Alfred James (Jim) Pittard was born in Ballarat in 1932. He completed secondary school at the Ballarat Church of England Grammar School in 1949. Pittard then enrolled in a diploma of pharmacy at the Victorian College of Pharmacy (1950-54), where he was apprenticed firstly to Cornell’s Chemist in Ballarat and then a pharmacy in Brighton, Melbourne. After graduation, Pittard worked for a year as a relieving chemist in rural Victoria before enrolling in a bachelor of science at the University of Melbourne (1956-58). He then completed a master’s degree (1959-60), again at the University of Melbourne.
Pittard was awarded a Fulbright scholarship in 1960, which took him to the University of California, Berkley. A year later, Pittard’s PhD supervisor moved to Yale University, and he followed. Pittard’s PhD degree was awarded from Yale in 1963. He remained in the USA on a US Public Health Services post-doctoral fellowship for another year before returning to Australia. Pittard spent the remainder of his career in the department of microbiology at the University of Melbourne, where he was appointed firstly as a lecturer (1964-66), then senior lecturer (1966-70) and finally professor (1970-1997). Pittard was made professor emeritus at the University of Melbourne upon his retirement in 1998.
Interviewed by Professor Michael Hynes in 2011
I am Michael Hynes and I am here to discuss his life in science with Jim Pittard, a renowned molecular biologist.
Jim, you were raised in Ballarat, a large country town in Victoria. What are your memories of this and how it affected your subsequent life?
I think it was a pretty happy time, Michael. We lived in Crocker Street for most of it and there were lots of kids in the street that we could play with. I remember playing marbles, making shanghais, making bows and arrows, going fishing on the lake, paddling a canoe on the lake, playing cricket, kicking a football, playing baseball – after the American soldiers were in Ballarat – and riding my bike almost everywhere. It was really a time of great enjoyment.
You went to school in Ballarat. How did that affect subsequent events?
I went to primary school at Pleasant Street state school. It was considered by my family that you should go to the state school for primary. I have to confess that I don’t really recall a great deal about it. I do remember playing the triangle in the first grade. I do remember learning my times table in the second grade and I think in every grade after that, until the sixth. I do remember also some of the teachers. The teacher we had in grade 5 was a sadist, really. He had a leather strap that he folded over so that, when he hit you, you got two bangs: one from the first and one from the second. He also used to delight in making the students kneel on a little dais up the front of the room until they almost fainted. So he was not good news.
Fortunately, the sixth grade teacher was lovely – a very nice chap. So I recovered from the maltreatment in grade 5. I do recall in grade 6 that we had a visit from someone in the temperance society telling us all about the evils of alcohol. They had a tract that they gave us and they said that, if we studied the tract, learned it well and got it correct, we could get a pound. I remember that I learned those three pages. I can still say the first bit: ‘Alcohol is not a stimulant but a narcotic, an anaesthetic, a drug which paralyses first the willpower and then the other higher faculties of the brain.’ That was 70 years ago I learned that. I did the test and I got 99. I don’t think it turned me to drink immediately, but I was very disappointed.
The other thing I recall is the fiasco of air raid drills. It was at the time when Japan had bombed Darwin and people were getting slightly worried about whether they were going to bomb everywhere. The drill was like something out of a French farce really. When they rang the bell, the kids in grade 4 went into the grade 5 class and hopped under the desks, the kids in grade 5 went into the grade 6 class and hopped under the desks and the kids in grade 6 went and marched into the bottom of the empty swimming pool and sang God save the King. So that’s my recollection of my primary school.
Jim, did you feel at all scared because of the war?
No. It was a bit unreal. We had to tell them how we were going to get home, if we had to get home. I said that I was going to walk home around the lake and so on. We didn’t really understand what war was like.
So then secondary school.
At the end of primary, I won a small scholarship that let me go to the Church of England boys’ grammar school. It was the school that my father had been to and where my brother was currently a student. That was a small school with only about 160 students all told. But it had a quite inspirational headmaster, Jack Dart. He was a philosopher, a classics scholar, a person of great integrity and a very hard worker. It was during the war and Jack used to maintain an enormous vegetable garden that provided all the boarders with vegetables. He used to get up every morning at five o’clock and go and work in the vegetable garden and then do his teaching. The other person who taught me at that school was George Seddon. George taught me English literature in the last year. So the classes were small. In matriculation, I think we had between three and five students. The teaching was informal. I can remember that we had one teacher who liked to teach us logic as we went walking around the pine plantation. We used to do English literature with George Seddon sitting up in the vegetable garden. And we used to do physics and chemistry, with a dear old teacher who had been pulled back out of retirement, sitting in the big chairs in the teachers’ room.
The other aspect of the school being so small was that you were really involved in everything. In my last few years I was in the cricket team, the tennis team, the football team, I trained for athletics, I was rowing for a while, I was sergeant major in the cadets, I was in the school play and, in my last year, I was captain of the school. I took all this very seriously and it required a lot of effort. That was the good side of things. It was really enjoyable and I think very important. The bad side was that academically it was a bit of a downer for me. My matriculation results were disappointing. I passed English expression, English literature and British history – which another student and I taught ourselves – and I failed physics and chemistry.
In terms of a career in science, that wasn’t a very good start.
It is interesting to think back to those times. At that school at that time, matriculation wasn’t really the focus of education. Maybe the teachers were rebels, but they almost regarded matriculation as an intrusion. So, in answer to your question whether it was important for later life – yes, it was very important. I learned a lot about responsibility, about initiative, about integrity and ideas. In the last couple of years at school, I discovered poetry, I discovered ideas and I discovered originality. So, yes, it was. The down side was that, as far as qualifications were concerned, I wasn’t doing too well.
This led on to your choice of career. How did your family influence this? For example, your father was the owner of a shoe store, this was a family business. Did you ever consider joining the family business? How did the family influence your choice of career?
I never ever thought about the possibility of going into the shoe store, even though the Pittards for generations had been involved with shoes. I had an elder brother, and thank goodness that was his destiny. My father thought I should do medicine, dentistry or pharmacy. Since I had failed matriculation, that only left pharmacy. I had passed leaving chemistry, and that was sufficient. So I signed up for a fouryear apprenticeship with Walter Cornell and Son, Druggists, in Ballarat.
I started off on a wonderful salary of 26 shillings and sixpence a week. For the first two years I worked at Cornells and studied by correspondence. Then I went down to Melbourne, changed my apprenticeship over to someone in Brighton. I continued working in a shop and went to pharmacy college in the afternoons. I quite enjoyed the course in a way. I enjoyed the challenge of learning in the last two years. I think it gave me an opportunity to demonstrate that my non-achievement in matriculation was not a lethal event. In the final year of pharmacy, I got the Ramsay Pharmaceutical Prize in chemistry. I thought that wasn’t too bad for someone who couldn’t get matriculation chemistry.
After you got your qualifications as a pharmacist, where did you work?
When I qualified, I worked for the next year as a relieving chemist. This meant that I travelled around Victoria and took over shops in country Victoria, while the owners went off for their annual holidays. I travelled by train and bus because I didn’t have a car and I stayed in country pubs. The pharmacies varied. Some shops that I ran were quite a challenge because they were busy and there were lots of things to do. I quite liked making up prescriptions, making ointments and mixtures. I got a bit fed up with counting out tablets and scraping off labels, but the dispensing I quite liked. But mixed in with the busy shops were some shops that had very little trade at all, and that was absolutely killing. I hated it. It was so boring. You would go to work and you simply had to wait for someone to come into the shop to give you something to do. You couldn’t exercise initiative. I decided when I did that, ‘This is not for me, and I’m not going to spend the rest of my life doing this.’ So I started to think about what I could do. I wanted to have more education, and science seemed to be the logical thing to do. For a while I toyed with the idea maybe that I might do science and set up a pathology lab next to my chemist shop. But, in the end, I didn’t do that. I knew, if I were to do science, I would have to do physics, chemistry, maths and zoology. So, during that year when I travelled around Victoria, I used to sit in country pubs at night with a physics book and a maths book, trying to bone up on those subjects, which was not easy, I must say.
But you did get into Melbourne University to do science.
I did get in. Today they wouldn’t let me in the back gate.
It has become so competitive these days. In telling your family about this decision, what did they think?
They were not pleased, Michael. By this time, Barbie and I had become engaged. I recall a discussion with my father when he pointed out at some length the onerous responsibilities that I was taking on. Which he didn’t think were met by planning a new course in science. My dear old gran, of whom I was very fond, took me aside and said, ‘Are you sure you’re not giving up the substance for the shadow?’ I assured her that I was not. They were not too happy with that, but that is all right. Barbie was very supportive and that was the main thing.
You finished your science degree and then made a decision to start a master’s degree working with bacteria.
While I was an undergraduate, I used to work at Braithwaite’s pharmacy out in Camberwell on Wednesday afternoons and Saturday mornings. Barbie was working in St George’s Hospital at that stage. Then, when I finished my science degree, Frank Gibson invited me to join his lab to do a master’s degree. Frank was at Melbourne and was a fantastic scientist and a great fellow. I hadn’t really thought about it, to be honest, but I was absolutely delighted. Frank was able to offer me £700 a year as a stipend. I can still remember my father shaking his head and more or less saying, ‘Son, you’ve done four years pharmacy and three years science and now you’re being paid £700 a year?’ He couldn’t understand it. Anyway, we were delighted. It was a great opportunity. So, I started doing a master’s with Frank.
When did you first meet your wife and what were your early married years in Melbourne like?
We met when we were still at school. We were both in Ballarat and we used to meet occasionally. We were very taken with each other. It was only during the time I was working as a pharmacist, when I first finished, that we really got to know each other properly. We got engaged at the end of that year and then we got married at the end of my first year of science. The first year of science I had in Trinity College and then we got married. We then rented accommodation around Melbourne for three years. It was a good time, really. We were poor, but everybody’s poor and we had a good time. Christopher was born towards the end of my science degree, in 1958. We had managed to get a small house built at Montmorency, so we moved into that when Barbie and Chris came home from hospital.
You already had a house in Melbourne when you were doing your degree.
With the master’s, yes.
So you were fairly well established. Overall during your career, has easy has it been to manage your family life and the demands of your career?
You should probably ask the family. It’s difficult, I think. The problem is, if you are teaching and doing research, it is a very demanding job and you do tend to be very preoccupied. Particularly with research, you spend lot of time thinking about problems you are trying to solve and you are very busy. I think people in that situation are very conscious of how much effort they spend trying to spend more time with the family. But you need to ask the family whether that effort is satisfactory.
Sometimes you can physically be there but mentally not.
That is right. It is constantly in your head. But we went overseas to do a PhD and then we had a period of travel. So it was only when we came back, that we really had to struggle with these demands.
After you’d finished your master’s, which took two years or so, you decided to do a PhD in the States.
I started with the master’s and I was working in the chemist shop every Saturday morning to get some money. Then Syd Rubbo, who was chairman of the department of microbiology, asked me whether I would be senior demonstrator. So, while I did my master’s, I was also senior demonstrator preparing all the material for class and so on. I don’t think you are supposed to do that, but I needed the money and Syd needed a senior demonstrator. So I did that for two years.
Then, when I had finished the master’s, Syd called me into his office. I noticed that he had this big book on The microbial world that had just been published by Stanier, Doudoroff and Adelberg, who were at Berkeley. Syd said to me, ‘Why don’t you go overseas and do a PhD?’ and I said, ‘Oh yeah, that sounds like a good idea.’ He said, looking at the book, ‘Why don’t you go to Berkeley?’ and I said, ‘Oh, okay.’ He said, ‘You should work with Stanier.’ So I said, ‘I’ll write to Stanier and see what happens.’ Syd encouraged me to apply for a Fulbright Fellowship, which, to my surprise, I won. Syd and Nigel Manning - Nigel Manning was dean of the pharmacy college – got together and they also got me a stipend of £700 a year from Harold Woods. Woods used to make Relaxa tabs. These were tablets that you could buy over the counter. They were a fantastic success because everybody thought they were stressed and needed relaxing and you could get ‘relaxed with relaxa tabs’. Anyway, I got £700 year from Mr Woods for two years. The idea was that when I came back to Melbourne I would do some teaching at the pharmacy college, which I did.
So I wrote to Stanier, and Stanier wrote back, and said that he was very sorry but he was about to go on sabbatical leave to Paris and didn’t have a place. I was thinking of writing to Doudoroff and I got this very nice letter from Ed Adelberg saying that he was happy to offer me a position in his lab. In that way I became a microbial geneticist, because that is what Ed was doing. So that was it. We packed up and launched ourselves off.
Would you say that doing a PhD after you had finished your master’s was accidental?
I don’t know whether I was just living from day to day. I really was very busy, being a senior demonstrator and a pharmacist and a father and a student. So I really don’t think I was looking ahead. Obviously I was very happy to accept the advice. The notion was one that I was very interested in. But I think it was not until I was doing PhD work in America that I felt like a full-time research person, because I was too busy doing too many things.
Obviously going to the States was a big experience. Perhaps you could tell us about how you reached that decision, your experiences travelling there and your experience then as a PhD student in the States.
The travelling was a great experience. The only way to get to America in those days was by boat, so we travelled on the P&O liner Himalaya. Fortunately for us some friends of ours, Barry Egan and his wife, Janine, were travelling on the same boat. Barry was going to Denver to do a PhD so we spent a lot of time together on the boat. The voyage was a sort of mixture of excitement and boredom. The excitement was all the stop-offs. We stopped at Manila, then Hong Kong, then Kobe and then Yokohama. After Yokohama, we went to Hawaii. From Hawaii, we went up to Vancouver. Then from Vancouver we went back to San Francisco. They were the high points of the trip, without any doubt. In Manila we went out at night and watched them play jai alai. It was good fun.
The low points were when we started off. Christopher - who was 2½ - had a raging fever when we first got on the boat. We weren’t too sure how he would be. We thought that perhaps they weren’t going to let us on. Anyway, after a while he was okay. We left Japan just before a typhoon got in, which meant that we had five days of the most horrendous weather. I don’t think I ever want to experience that again.
Were you seasick?
I was absolutely seasick. Barbie was terribly courageous and heroic. At one stage she dragged me right up to the top deck, where we shouldn’t have gone, to look at things. You looked out and the seas were up there (indicates) somewhere. How the ship kept going I have no idea.
We got to San Francisco, got to Berkeley, started looking for accommodation and had absolutely no luck at all for two weeks. We were shown dreadful little apartments miles away, nowhere near the campus. Then one morning I went up and I really believe that they got my file mixed up. They saw ‘Fulbright’ and they thought that I was some visiting Fulbright staff member. All of a sudden they showed me a list of places that I had never seen in my life. Five minutes later I had grabbed the downstairs of a house that was only two blocks from the campus. I signed up and we were there. We were in California for a year. We bought a great second-hand Ford station-wagon with little curtains on the windows that we used to go travelling in. We went to all the national parks –Yosemite, Sequoia, Death Valley – and had a great time.
The first semester I was there, Adelberg said that he didn’t have any room in his lab for me to do any work, which was a bit of a surprise. He said that they weren’t too sure what my master’s really meant, so would I take some graduate courses? I had to take two graduate courses for credit, which were enzyme chemistry and immunochemistry, and I had to audit three others. I did that quite satisfactorily. In the second semester there was room in his lab, so I went into the lab and started on my research topic. I worked as a teaching assistant in immunology at the same time.
At the end of that first year, Adelberg then said, ‘I need to talk to you. I’ve got some news.’ I said, ‘What’s that?’ He said, ‘I’ve been offered and I’ve accepted the position of chairman of the department of microbiology over at Yale University over the other side of the country.’ He said, ‘You’re very welcome to come with me, if you’d like to. I’d be very pleased if you would come. Otherwise, you might like to think of changing to someone like Pardee.’ I was thinking about Pardee, but Pardee had just been appointed to Princeton, so he was just moving on. Barbie and I talked about it and we said, ‘Sure. We’re only too happy, we’ll come to Yale.’ So at the end of the first year, we packed up and we drove down to Pacific Grove, where I was taking a fantastic course in general microbiology given by Klaus van Niel. Then we drove to Denver and stayed with the Egans for a couple of days. Barbie wasn’t too well, so she and Chris flew to Michigan to see some friends and then from Michigan to New York. I pointed the Ford in the direction of the east coast, drove across to New York, picked them up and took them back to New Haven.
For the first four weeks in New Haven, we lived in a little quanset hut. They had these army quanset huts on the polo fields and they had their graduate students living in them. They had a coke-burning stove inside to keep you warm in winter and they had hoses on the roof to cool you down in summer. I remember that my parents visited while we were in one of these. I think my father thought his predictions had proved correct. After that, we got into some new apartments for married graduate students. We stayed there for three years and that was fantastic. It was a very interesting group of people. Still on the travel, while we were on the east coast, we went up to Vermont many times. They were interesting trips. The Egans came across from Denver and we drove with them up to Canada, went around the Gaspe Peninsula and then came back. That was all really very interesting. I mentioned before, we had a station-wagon. We put a mattress in the back. Barbie and I used to sleep on the mattress and Chris used to sleep on the front seat.
Did you find American science rather different from what you had previously experienced in Australia?
I enjoyed it. It was fantastic. It was a very exciting time to be there. All the work in France – Jacob and Monod’s results – were coming out. There was a lot of work going on.
Perhaps you would like to explain about Escherichia coli that you first started to work on in the States.
I had been introduced to Escherichia coli and Aerobacter in Frank’s lab. But E. coli was really a fantastic organism. It had a lot going for it. First of all, it was non-pathogenic, which was a good thing. Secondly, it was very easy to grow. You could start off with one and finish up with one thousand million cells after overnight cultivation. It was an organism that had a fantastic synthetic capacity. In other words, this organism would grow on a simple sugar like glucose and ammonium salts. It made everything else it needed. All its amino acids, all its nucleotides, all its vitamins, it could make from these simple building blocks.
Also, because it divides by simple fission, the thousand million cells that you grow overnight you can really treat as a single individual in a way. This means that, if you want to look at enzymes, the fact that it is a tiny little organism – only one-micron long - doesn’t make any difference. You can work with a thousand million of them and you can extract the enzyme and you can see exactly what is happening. There were well established methods for extracting enzymes, for breaking cells.
In the late 1940s, Lederberg had discovered sex in bacteria, which again involved E. coli. He had established that the bacteria could transfer genes from one to another by a process called ‘conjugation’. In the late 1950s, people like Jacob, Wollman and Bill Hayes in the UK and Adelberg in the States had all worked on this conjugation system. They had managed to make it into a very efficient system for transferring genes from one cell to another. Not only was it efficient but also it was a system whereby you could do mapping very easily to find where the genes were on the chromosome. Norton Zinder and Lederberg had also discovered that you could do transductions. That is, that bacteriophage (bacterial viruses) can pick up part of the chromosomes and take it and put it into another cell. So all of the techniques were there for working with E. coli. Actually, in the 1960s, it is probably fair to say that more was known about the chromosome and the genes of E. coli than any other living organism.
Oh yes. I’m absolutely sure of that.
It continued throughout the 1960s because it was studies of E. coli that contributed to the cracking of genetic code and working out how the nucleotide sequence represented amino sequence in proteins. It was also the discovery of messenger RNA. You would have to say that E. coli was also involved in the birth of molecular biology. You can attribute many of the major discoveries to work with E. coli. So why would you not want to work with it?
That’s right. You actually got turned on by it and were really excited when you were in the States.
It was great. Adelberg had come back from a year at the Pasteur Institute. He had gone over there on study leave and his intention had been to work with Georges Cohen on isoleucine valine biosynthesis. This is something that he had worked on earlier on in his life. He worked with Georges for six months. But, at the same time, in another part of the building, Jacob and Wollman were busy doing all the conjugation work and all the work with operators. So Adelberg then politely moved from Georges Cohen’s lab to Jacob’s lab. Adelberg published that paper with Jacob on the too-early-interruption easy way to make F-primes. So he came back from the Pasteur all fired up about F-primes, operons and operators – all of this. It was exciting times. But it also was exciting because everything was happening. People were trying to crack the code – they didn’t know what it meant. There were interesting people with interesting ideas trying all sorts of way-out experiments. It was a great time to be in science actually.
I was an undergraduate then and what was happening then was really exciting. Your organism has always been E. coli. Did you ever consider changing to another organism – for example, an animal system?
No, never. The reason is very simple. Once we got started on our various projects that we worked on, we always had more questions to answer than we had time, students, money or anything else to put to them. And you do get pretty obsessed about trying to find the answers to some of these things.
And you were getting grants and being supported to do that, so why change?
That’s right. The reality also, even in those days, was that people who were very successful in one field, would go to a new field and apply for grants and all the reports would come in and they would say, ‘What’s this person done in this field?’ It was very difficult. If you didn’t have a track record, you just didn’t get there.
After you had finished your PhD, you finished up working at Melbourne University for your entire career doing both teaching and research. In addition, you were head of department frequently during that time. How difficult was it juggling all of these demands?
It is a bit tricky. I really enjoyed teaching. I think it was very rewarding. It is emotionally draining and a lot of work. But I liked it and I enjoyed both my undergraduate teaching and, in particular, I very much enjoyed teaching graduate students. The research and the teaching, in that sense, were very much combined, because much of my research was carried out by graduate students. I think this is often the reality. If you are an academic in a position where you are doing teaching as well as research, you do not have a great deal of unbroken time yourself to spend in the lab. So you do a lot of it through the PhD students who are working for you.
Administration: look, I was lucky with regard to administration. The microbiology department, ever since Syd Rubbo’s day, had always had the tradition of employing a ‘lab manager’, now a ‘business manager’. Who was very efficient and excellent person in that job. We had Jim McEwen for many years and, later on, John Gorry for a number of years. That person had an absolutely vital role to play in running the department. I used to say when I was head that, ‘if there were a day on which I didn’t come in, noone would notice. If there were a day on which John Gorry didn’t come in, the whole thing would fall apart.’ He would manage all the finances, hiring and firing of technical staff – all sorts of things. This meant that, as head, I was relieved of a lot of the administration that I have seen really bog my colleagues down. I have seen some people who really just get absolutely smothered in this stuff.
The second reason I was lucky was that the late David White and I had an arrangement whereby we rotated the headship. So each one of us would be head for about three years and then we would swap. Three years you could just about manage. You could still keep pretty much in touch with what was going on in research and get back to it. When I was appointed head of department the first time, I spent the first four weeks, in the head’s office, which was down on the first floor. It was a bit like being in the pharmacy again: I hated it because the doors were closed and no matter how hard I tried to get out of there, I was being locked in there more and more. So I just got out. The admin secretary was a little bit astonished, but I said, ‘I’m not going to stay in there. I’m going back to my office next to the lab. But, every morning when I come in, I’ll come and see you, get the mail, take it upstairs, deal with it and come back and give it to you.’ So I did that. I lived up next to the lab, which was great. It meant that I always had the door open and I could move into the lab whenever I wanted to. The students could come in. I always had an opendoor policy: people could come in whenever they liked. Those elements were really important in allowing me to cope with things.
As far as the university was concerned, I spent a lot of time in the early years on faculty of science committees. In later years I restricted myself to things that I had to be on: the medical executive board, the faculty board, and research and graduate studies. I concentrated on the research area and tried to stay out of other aspects of administration.
You worked for Melbourne University for your entire career. Was it a good employer? How have things changed at Melbourne University? Would you like to compare it now to how it has been during your time?
It’s difficult for me to comment much at this stage, Michael. It is certainly a very different place. It is so much more complex, there are so many more students now and lots of overseas students that we didn’t have before. The facilities are much better, in general. The workload seems to be pretty heavy on staff at the moment. They seem to be pretty stressed with not only the workload but also, the pressure on people to achieve. The pressure on people to be tops in teaching and research is pretty relentless. I can’t see the department currently taking off for a game of cricket with the department of physiology, like we used to do. Nor do I see my colleagues taking four weeks annual leave to go and sit in the sun. I think it is different from that point of view.
It is different in many other ways too. The impact of electronic media is incredible, in terms of accessing information and communicating. When I started off, the journals that we got at Melbourne all came by boat. So you got the latest information about three months after everybody else. In actual fact, that was quite important. When I would first plan out our research projects, you had the feeling that you needed to have a project that was comprehensive enough so that your lab would be contributing most of the information. If you were going to be dependent on overseas, you were just not going to get it in time.
So, I don’t know. The Melbourne model has been going and it is bedded down now. It will be interesting in a few years to look back and see how successful or otherwise it has been. I don’t know.
When you returned to take up your position at Melbourne, how did you and why did you choose the particular research projects that you did?
Okay. In 1961, Jacob and Monod published a big paper in the Journal of Molecular Biology, talking about genetic analysis of regulatory mechanisms in bacteria. It was a paper in which they described the operon model about gene regulation. It was a very important paper. It created a paradigm shift in the way that people thought about how genes were expressed and regulated in bacteria. In their model, what they postulated was that there were two new genetic elements that one had to consider. The first one they called a ‘regulator gene’ or ‘repressor gene’. They postulated that this made something – they weren’t sure whether it was RNA or protein – which was expressed in the cytoplasm of the cell. They thought that that repressor was then able to attach itself to the second genetic element, which they called an ‘operator’. The operator was always located right next to the genes that were being controlled. So here is the model. There is a gene somewhere in the chromosome. It makes something called a ‘repressor’, which binds on the operator and stops those genes from being expressed.
The next part of the model is that small molecules, like lactose or tryptophan, can combine with specific repressors and change their activity. In the case of the lac operon the genes are switched on in the presence of lactose. So the model said, ‘The repressor binds the operator and stops it expressing’, but when lactose is there, that binds the repressor and inactivates it so that you get it switched on. In the case of the tryptophan pathway, the tryptophan genes were switched off by tryptophan. So they simply modified the model to say, ‘The tryptophan repressor is unable to act until the tryptophan combines with it. Then, when the tryptophan combines with it, it sits on the operator and switches things off.’ So this was their model. Once that was published, people all around the world went rushing off to their own systems to apply this theory to see whether it applied to their system – and I guess I was one of those.
So I came back to Melbourne. Frank Gibson was still there. Frank had been elected to the Australian Academy of Science by then for his work on identifying the branch point in aromatic biosynthesis, chorismate. He was subsequently elected a Fellow of the Royal Society. The pathway I was interested in was the biosynthesis of aromatic amino acids – phenylalanine, tyrosine and tryptophan. It is a complex pathway. It has one set of about seven or eight common reactions, leading to this compound that Frank had identified: chorismate. Then there are terminal pathways going off: three to the aromatic amino acids and four to so-called aromatic vitamins: folic acid, ubiquinone, vitamin K and enterochelin. Frank had Dick Cotton working on enzymes in the phenylalanine and tyrosine pathway; and, with Graeme Cox, he was starting to work on the pathway to ubiquinone.
Frank is in Melbourne and I am in Melbourne. Here is someone who knows all about aromatic biosynthesis. So there is a lot of expertise available. The genes were not properly mapped. Tryptophan certainly was done. Charlie Yanofsky had been working on the tryptophan pathway genetically and biochemically for some time. But the phenylalanine and tyrosine pathways and the common pathway had not been looked at extensively from a genetic point of view. There was some biochemistry done and a little bit of genetics. So we thought, ‘This is a good place to start.’ Brian Wallace was my first PhD student. Interestingly enough, he had also done pharmacy, before he started science, and had also worked at Harry Braithwaite’s. He and I started work and the first thing we did was to isolate lots of aromatic mutants.
Actually, let me tell you what techniques we had available at this time. We had mutagenesis, so we could get mutants. We had conjugation and transduction, so we could map the mutations. We could make diploids, so we could do cis-trans tests to see whether we had operators or regulators. We could purify proteins. We had radioactive amino acids, so we could measure transport. That was it. They were the tools.
So we started isolating mutants. We isolated lots of aromatic mutants and mapped them. It was pretty simple to find out where they were. We had some challenges. The first reaction in the pathway is carried out by three separate isoenzymes, which means that it is not easy initially to get mutants. If you knock one out, you still have two left that will carry out the reaction. But, fortunately for us, Colin Doy, Keith Brown and others had shown that each one of those enzymes is inhibited by a different amino acid. One is inhibited by tyrosine, one by phenylalanine and one by tryptophan. So we were able to use that sort of information to design moderately clever but not too complicated methods for isolating mutants. We knocked out those first enzymes one after the other until we had knocked them all out. Then we constructed a strain that had only the tyrosine inhibitable enzyme for that first reaction. We had shown with our mapping that the gene for that was situated right next to the gene of the first enzyme in the tyrosine pathway. Those two are sitting together, so they are good candidates for this sort of operon model. Other people had shown that they are repressed by tyrosine and de-repressed if you starve for tyrosine.
So how do we isolate this regulator mutant or look for it? Well, the strain that we had made, which just had the tyrosine-inhibitable enzyme, couldn’t grow if you added tyrosine to medium – because it knocked that out. The cells still needed that enzyme to make phenylalanine and tryptophan. Nor could it grow if you added a tyrosine analogue like paramino phenylalanine, which mimics tyrosine as a co-repressor but not as a feedback inhibitor. So we made resistant mutants. We mapped the mutations. Some of them, as we predicted, were closely linked to aroF, and they were clearly operator mutants. But we also found a whole bunch situated elsewhere on the chromosome in a gene that we called tyrR. That was our big discovery. We had found our putative regulator gene.
Helen Camakaris isolated temperature-sensitive mutants of tyrR and amber-suppressible mutants of tyrR, which showed conclusively that the tyrR product was a protein. She also made a lac fusion with tyrR and showed that it regulated its own expression. So, having got the tyrR mutant, we then set about finding what were the other genes in the pathway that were regulated by TyrR. I guess we spent the next 10 or 12 years identifying a total of eight different transcription units, or genes, whose expression was regulated by TyrR. Some of these were repressed. That is, you added tyrosine to switch them off. Some of them were activated. You added tyrosine or phenylalanine and expression went up.
At that stage, Michael, you would say that we had more or less come to the end of this project. There was nothing much else that we could do. Except, there was a lot that we could do! We had been sufficiently slow about doing it that gene cloning techniques, DNA sequencing and a whole swag of new technologies were available. So we really were just at the beginning of something. We had all these genes that were regulated by TyrR and now we could clone them, we could sequence them and we could look at the upstream regions. We could have a look at the promoter sequence and we could identify TyrR binding sites. We could clone the tyrR gene and make up lots of TyrR repressor. Then we could use the repressor in purified systems and so on. So we were off and away.
It was a second burst of activities.
It was a second. For the next 10 or 15 years, we were busy and that is what we were busy doing. It is not easy to explain, but I will take this one example to see whether I can explain the sorts of things that we did. One gene which is regulated by TyrR is the tyrP gene, which codes for a transport protein that brings tyrosine into the cell. In the presence of tyrosine, this gene is switched off. All that means is that, when you have got a lot of tyrosine in the medium, some gets into the cell and once the cell pool is high enough, it doesn’t need to bring any more in. So it switches off the transport protein. It stops making it. If there is no tyrosine inside, if the pool is low, the transport gene is switched on to try and grab any tyrosine in the medium. If you don’t give it tyrosine but you give it phenylalanine, the gene is activated. They make more of it. The reason for that probably is that the cell likes to balance these similar amino acids in the cell. If there is too much phenylalanine in the cell, it is opening up the tyrosine transporter to try to grab in a bit more tyrosine to bring it in at the same time.
How does this work? Both these things are affected by TyrR? When we looked at the region upstream of the promoter of tyrP, we could identify two tyrR ‘boxes’. As a result of mutation studies, we were able to identify where the TyrR protein was binding. The tyrR box relates to a palindrome – you know this thing: ‘able was I or I saw elba?’ It goes back to front. The tyrR box sequence was TGTAAA, then six bases and then TTTACA. That is the ideal tyrR box. We looked upstream of the promoter and, overlapping the minus-35, there is a tyrR box and then, three bases away, there is another tyrR box. The upstream box is very much like the consensus. The downstream box, the one that overlaps the promoter, has a few mismatches in it. It has a few GC pairs in the central region, which the tyrR boxes don’t usually have.
By then, we had also purified TyrR protein. Actually, there were also then wonderful systems with gel and electrophoresis for looking at DNA fragments and looking at what is happening and using radioactive tracers. So we could add TyrR protein with tyrosine and with phenylalanine and look at what was happening to this region. We could show that, in the presence of tyrosine, both boxes were occupied by TyrR. Barrie Davidson’s lab, over in biochemistry, had shown that in the presence of ATP and tyrosine, TyrR was a hexamer. In the absence of tyrosine, it was a dimer. We showed that if you look at what is happening in the presence of tyrosine, you get both boxes occupied. The polymerase can’t sit on the promoter, because TyrR is already sitting there. So you get no transcription. In the presence of phenylalanine, only the top box is occupied. So the dimer sits on the top box and it can now interact with RNA polymerase sitting on the promoter to help it initiate transcription.
Arna Andrews and Blair Lawley then took this system and started inserting DNA between the boxes. So we could move the boxes apart and we could move the boxes further away from the promoter. We could ask the question, ‘What is the requirement here with these boxes? For example, do they have to be on the same face of the helix for repression?’ They showed that if you move these boxes apart, putting more bases in between them, repression disappears. But, after you have put 10 bases in between them, repression comes back. What that means is that those boxes have to be on the same face of the helix for the hexamer to bind across. Similarly, with activation: if you moved the strong box up or down, you could show that there was a face specificity of the helix for activation to occur. So that is the sort of thing that we were able to do at this stage. That is what we did with all of the genes. We wanted to understand how it was that this protein managed to use different amino acids to do different things to different genes. The interesting thing about having a ‘regulon’ is that there are subtle differences in the regulation of each one of these transcription units, and those subtle differences relate very much to the function of the gene which is being regulated. A ‘Regulon’ is what we call this system because you have got eight different transcription units controlled by the one repressor.
In 1991, I wrote a review with Barrie Davidson on the TyrR regulon. Basically saying what we knew about it at that time. That had just been published when I got a letter from Akira Ishihama in Japan. Akira said that he had read the review and was very interested. He wanted to know if we were interested in collaborating with him. That was great. That moved us on to phase 3. Akira was technically superb, there is no question about it. We got an ARC AustraliaJapan collaboration grant that gave us a little bit of money to travel. Blair Lawley went and worked in Akira’s lab for about four weeks. Yang Ji went and worked in Akira’s lab for about two months. Peixang Wang went and worked there for about four weeks, followed later by Shan Hwang. They got very important techniques working there and, what is even more important, they came back with fantastic material. The most important thing for us was RNA polymerase. RNA polymerase is a complex enzyme made up of a number of subunits. It was available commercially, but nearly everyone you spoke to said that it didn’t work. The enzyme you got was no good. Akira made his own and it was wonderful. Whenever anyone had anything to do with gene activation and RNA polymerase, Akira was there.
Anyway, Akira provided us with RNA polymerase and this then allowed us to do a whole swag of new things. Now, with RNA polymerase and purified TyrR, we could start doing in vitro transcription studies. Now we could take the DNA template, we could add RNA polymerase, we could add protein and we could see how they interacted. We could understand what, in transcription initiation, TyrR was controlling. It was also the ability to carry out this in vitro transcription that let us solve the problem of aroP. aroP encodes a general transporter of the three aromatic amino acids. The aroP gene is repressed by TyrR and by each one of those aromatic amino acids. That was a dilemma for us, because everywhere else the only thing that worked was tyrosine and phenylalanine. We never got TyrR-mediated repression with tryptophan. If tryptophan was acting, it needed the TrpR repressor, and Blair had shown quite clearly that wasn’t happening with aroP.
Once we could do in vitro transcription, Yang Ji and Peixang discovered that, in addition to the transcripts that made AroP, there was another transcript that went in the opposite direction. I will always remember the day I came into work and Peixang was standing by my office door. I came in and he said, ‘Prof, I think I have made a discovery.’ I said, ‘Good on you, Peixang can we talk about it?’ He had made a discovery. In actual fact, the way in which TyrR represses aroP is that it activates a promoter on the opposite strand. We knew that tryptophan would work to help it activate. So either tryptophan, tyrosine or phenylalanine activates the polymerase to bind this promoter on the opposite strand. When it does that, it blocks the one going in the direction to make AroP. This one on the opposite strand is a dud one anyway. It is a very tight binding promoter, but it doesn’t make anything. So that is phase 3.
Phase 4, the last phase, also involved Akira. We now had the sequence of the whole chromosome and we knew what tyrR boxes were. Actually, people in the States had scanned the chromosomal sequence and identified where there were various tyrR boxes on the chromosome. We used a slightly different mechanism. Akira called it Selex. How it works is as follows. You take the E. coli chromosome and break it up into fragments. Then you mix those fragments with TyrR protein under appropriate conditions. Any fragment that has a TyrR binding site will bind to the protein. Then you separate out the protein with the fragments bound to it. You take the fragments off the protein and use PCR to amplify them. Then you can sequence them, and then you can use any one of all the wonderful IT things that are around to tell you where that comes from on the chromosome. Then you have a list of a number of genes that have tyrR boxes associated with them. We had eight or nine genes where the tyrR boxes looked as though they might be doing something because they were just upstream of where the gene was. We went through a quite exhaustive study in which we made lac fusions and studied regulation of these genes. I regret to say that, of the nine, only one – that is the folA gene – could we show was definitely regulated by TyrR.
That was even though they were in the right sort of position.
Even though they were in the right spots! Anyway, folA worked out to be a new member of the regulon. The others – well, I’m cautious. I would have to say that we failed in our attempts to demonstrate that they were part of the regulon. We couldn’t demonstrate that. But it may be that we just didn’t have the right conditions.
Organisms are always smarter than we are.
That’s right. Over the years we have mutated the tyrR gene and have identified regions of the amino terminal domain. In particular Yang Ji has done a lot of this work, and Helen Camakaris also. It is a big protein. We have two patches of the amino terminal domain that are necessary for activation: one which binds the amino acids and one which interacts with RNA polymerase. We have got mutations in the central domain which affect hexamerization, ATP binding and so on. In the carboxy-terminal domain, we have identified clearly the DNA binding domain and which bases interact with which amino acids in the protein. It is a very well-studied protein at this stage. Helen, and Tadeshi Fuji, also managed to identify two of the amino acids in the alpha subunit of RNA polymerase that interact with TyrR. So we are almost there.
Okay, plasmids. The second major project we had was plasmids.
Did that begin when you first came back?
Yes. When I was in the States, there was a lot of interest at that time in antibiotic resistant plasmids in Japan. Lots of strains – shigella, salmonella – were turning up that were resistant to four or five antibiotics. People were very worried about it. Then it was shown that these antibiotic resistances were sitting on plasmids. Plasmids are little mini chromosomes that were also able to transfer themselves by conjugation from cell to cell. I had an abiding interest in plasmids. My PhD work had been with F-genotes and conjugation.
When I came back, there had not been a great deal of work done on plasmids in Australia. So for the first two or three years, we did pretty straightforward epidemiological experiments. We were looking at organisms from hospital outbreaks of gentamicin-resistant strains of Klebsiella. We were looking at the organisms carried by refugees who had recently come in from Vietnam. We were looking at the resistance plasmids that you might find in organisms that you got from cattle. There was a big question about whether or not there was transmission of these things from animals to man and back again. So that is what we were doing for the first few years.
You couldn’t do a great deal with plasmids at that stage. You could determine the antibiotic resistance phenotype easily. You could determine something called ‘incompatibility group’, which simply meant, if you had one of those plasmids, you could test whether it could coexist in the same cell with certain other plasmids that had been identified. You could measure them and we started off measuring them, but that really was enormously tedious. You had to extract the DNA, separate the plasmids from the chromosome, purify the plasmids, put them on planchettes and put that in an electron microscope. Then you could maybe measure and see how big it was. We did that to start off with, before gels and things. It was very hard work.
We were using the incompatibility test quite a lot. We discovered that a lot of the plasmids that we had, seemed to have more than one incompatibility locus. They were complex. Furthermore, the plasmids were very big. They had lots of antibiotic resistance genes, conjugation genes and so on. They were hard to work with. So we thought, ‘Why don’t we use the gene cloning techniques to make little ones? We just need the genes for replication and nothing else. That little plasmid should be able to replicate quite well in cells.’ We also put in the genes for galactose catabolism and then we had something that we could measure easily on a plate. We made up a whole swag of things we call mini plasmids. We were using those to type unknown plasmids. We sent them out to quite a few labs around the world. But then, by this time, technology had improved and now you could get radioactive probes and you could do it more easily that way. So we stopped doing that.
Yet we had these mini plasmids and we thought, ‘Let’s take a group of these and use what we have got to ask the question, ‘How is replication regulated? How do these little plasmids regulate their own replication?’ This then became the big project in the plasmid area. We worked on the Icomplex plasmids. I should point out that the regulation of plasmid replication is very different from TyrR regulation. In the case of TyrR, when the cell is growing in the presence of tyrosine, it is not making any enzymes. You transfer it to a medium where there is no tyrosine, and now it has to make those enzymes and it has to make them quickly because it is in competition with all sorts of other cells. So, with the TyrR control, you will get more than a 100-fold increase in the rate of synthesis very quickly. In the case of replication control, these plasmids control their replication so that there is only between one and three copies of a plasmid per cell under any circumstance. There’s got to be more than one so that, every time the cell divides, each daughter cell gets a copy. But it costs a lot of energy to make these things, so the cell that has a big copy number is at a disadvantage to the cells that don’t. With the fine-tuning of this regulation, it is a different system to TyrR. Furthermore, the other difference is that, in the case of these plasmids, the regulation is affected at the level of translation: and not transcription. TyrR affects the transcription but with these plasmids, control acts at the level of translation.
I will refer to this diagram (indicates), which will make it easier to explain. I won’t go through the work but just give you some idea of the complexity and the elegance of this system that we finally discovered. The critical protein being regulated here is called a ‘RepA protein’, and that’s responsible for replicating the plasmid. The messenger RNA, which involves the coding sequence for RepA, has a long leader sequence to it and it forms paired structures, like this one here (indicates). The interesting thing about the paired structure which is formed with this RNA is that the start site and the ribosome binding site for RepA are locked up in it. They are not accessible to ribosomes, so nothing is going to happen.
However, in front of the RepA transcript, there is a region which codes for a leader peptide called ‘RepB’. So, when a ribosome comes and translates repB, as it comes along translating this message, it opens up this paired structure which reveals the binding site and the start site for repA. First of all, you get translation of repB and that makes repA ready to be translated. When that happens it allows a reaction to occur between another loop in the RNA here (indicates). This can form a structure called a pseudoknot. So we have a pseudoknot which is just upstream of the ribosome for repA. If you don’t get pseudoknot formed, the ribosome won’t work either. The ribosome that is translating repB comes along here (indicates) and stops, and Judyta and others have shown that this ribosome is the one that comes back and translates repA. Nothing else can get in here (indicates). I should say that Judyta Praszkierr must have credit for most of this work. There is no question about that. So that is the simple solution. That is how repA is translated.
However, it doesn’t happen like that. There is an antisense RNA which is made which can combine with this stem- loop here (indicates). When it does, as in this case, the pseudoknot cannot form. When the pseudoknot cannot form, translation can’t occur. So the small antisense RNA is controlling what is happening. You have the balance between these things. Basically, you have a series of regulations that have come together to give you this very fine structure control.
We spent many years on this and did lots of mutational studies. Ian Wilson made many mutants. Kirby Seimering did some beautiful work on the interaction between the antisense RNA and this RNA here (indicates).
That is interesting because the use of small RNAs in eukaryotes has been widely acclaimed in recent years, but it has been present in bacteria for a long time, including your discoveries.
And a lot of very interesting work has been done about how the RNAs interact and how they control things. There is no question about it. So that was the second major project we did.
We also worked on the transport proteins. When we found that they had been regulated by TyrR.
You had a little bit of involvement with the commercial world in terms of the biosynthesis of tryptophan.
We had all these mutants, so we thought maybe we would have a go at seeing whether we could make something. Tryptophan was an amino acid that was required commercially as an addition to stockfeed. It is one of the amino acids that are very low in things like sorghum. So we set to try and make strains that would produce tryptophan. We were successful enough to do a deal with a big German company called Degussa. They paid – I’m amazed to say – a $1 million licence fee for our strains. They then supported our research for another four or five years after that. We had some success. We had strains that were almost commercially viable. But we had some problems with stability that I don’t think we finally solved. It was an interesting time, an interesting experience. I went to Germany a number of times and we had the Degussa people out in the lab. Eberhard Breuker came out and worked with us for quite some time.
In the 1970s, E. coli was at the centre of the developments in recombinant DNA and genetic engineering work, so you were invited to go to a groundbreaking meeting at Asilomar in California to consider the possible risks and dangers of this. Perhaps you would like to tell us about this meeting.
In terms of background to this, Paul Berg and the late Bob Symons had been planning to do an experiment with lambda and SV40. Lambda is a virus of E. coli and SV40, as you know, is an animal virus. They had planned to linearise these and use homopolymer tailing to make polyA on one and polyT on the other and join them together. They were then going to put this chimeric molecule into an animal cell to see what happened and to put it into E. coli to see what happened. Some scientists got excited about that and said, ‘We think SV40 might be oncogenic and, if you put SV40 into E. coli, is there a danger that you might make an E. coli strain that can induce cancers?’ So that experiment was not carried out.
Herb Boyer and Stan Cohen had discovered that, when you cut DNA with restriction enzymes, they make staggered cuts with sticky ends so that you could join cut molecules together. That opened the field up to all sorts of possibilities of mixing genes and putting them into cells. So a number of scientists wrote and suggested that there should be a moratorium voluntarily put on the work. Berg and others then organised this big conference at Asilomar to discuss potential risks and how they could be handled. There were 85 scientists from the States and about 35 from other countries who went there. Jim Peacock and I were there as delegates from the Australian Academy of Science. Bruce Holloway was also invited as a specialist in pseudomonas genetics. It was an absolutely frenetic 3½ to four days because of the way it was organised. The mornings were mainly taken up with people reporting on the latest experiments that had been done using this new technique. It really was quite mind blowing.
So there was science?
Yes. I can remember Stanley Cohen talking about taking some histone-coding DNA and putting it into E. coli and being able to demonstrate now there was some RNA that wasn’t there before. Herb Boyer was talking about some experiments they had done with staph plasmids. So the mornings were spent on the science and the afternoons were spent trying to work out what the risks were and how they could be handled.
The problem was that the risks were all absolutely hypothetical. In essence there were three groups of people there – three working parties, if you like. There were the microbial geneticists, who were concerned with bacteria and antibiotic resistance and things like that. There were the eukaryotic plant people and animal people, who were interested in those cells. And there were the virologists, who were more into the cancer side of things. The interesting thing was that members of each group were totally convinced that their own work was perfectly safe, but also they were equally suspicious of the work that the others were doing. There is a message in that. Really, what it says is that what people are frightened of is not what they understand, what they are frightened of is what they don’t understand.
Just to set the background to this meeting, the media was also invited. They also invited a number of legal people. I still remember clearly a lecture given by one of the law professors towards the end. Basically he was saying, ‘If you blokes don’t regulate this and do it properly, we’ll regulate you out of business.’ It was very interesting. Anyway, this was the dilemma.
There were a lot of very bright people there – Sydney Brenner stood out particularly, Roy Curtis III, a number of very smart people. So every time we considered different experiments, because we didn’t know the answers, people said, ‘Well, look, let’s consider the worst case scenario.’ What that means is that they say, ‘If you’re using E. coli as the host, let’s assume that the piece of DNA that you’re putting into E. coli will make it into a pathogen. What can we do to make sure that, if that happens, it won’t escape from the laboratory and nobody will get sick?’ They came up with a set of guidelines on physical containment and biological containment. The physical containment stipulated what the labs should be like and how the procedure should be done and the biological containment included all sorts of very clever plans about attenuated strains of E. coli and about plasmids that couldn’t be transferred. At the end of four days we had a draft set of guidelines for all these experiments as to how they should be carried out. There were certain experiments that were prohibited by that meeting. For example, it was decided that you shouldn’t put genes encoding toxins, like tetanus toxin, into E. coli, and that you shouldn’t release any modified organism into the environment.
Jim Peacock and I came back to Australia and gave a report to the Australian Academy of Science. The Academy of Science set up its own committee – the Academy of Science Committee On Recombinant DNA (ASCORD). Gordon Ada was the chair of that. For about five years, we oversaw the development of recombinant DNA work in this country. We produced guidelines. The whole system was voluntary, but we had agreement from heads of universities, from ARC, from NHMRC and from CSIRO that everybody would abide by the guidelines. It actually worked quite well.
After five years, the Academy commissioned another report. It was a review, with Frank Fenner as chairman of that review committee. Nancy Millis and I were on that committee. It is interesting to read the report on it. One of the things it said was that many of the hypothetical concerns that had been expressed five years earlier had turned out not to be valid. Nevertheless, it thought that regulation should continue and there should be a new committee. They wanted the committee to be government-funded. There were two reasons for this. One, although it is probably not written anywhere, is that the Academy had approached the government for 10 grand to provide secretarial assistance to the committee. The government had more or less told it get lost. It is interesting to note that, with the new government committee set-up, the budget in the first year was $85,000. Anyway, that is the first point. The second point was that it looked as though industry was about to get involved, and people are always concerned that these nasty industrialists and commercial people are going to do things. The feeling was, ‘Maybe we should have a government committee. Maybe that will have a bit more clout to control things.’ Then the Recombinant DNA Monitoring Committee (RDMC) was set up. Nancy Millis was chairperson of that and I was also on the scientific committee. That one went on for about another six years.
The government then decided that it wanted to change the committee again because now it looked as though agriculture was going to get involved. This meant that they were going to have to look at planned release. Up to that point, we had produced numerous guidelines on control. Having made that decision, the government took about two years to do something about it. We were in limbo there for quite some time. Then they produced the Genetic Manipulation Advisory Committee (GMAC), which had this wider purview of also looking at planned release. Nancy Millis was chairperson of that and I was chair of the scientific subcommittee of that committee.
For about 10 years, we handled all the experiments that came through. As chair of the scientific subcommittee, I looked at every application that was made in Australia. I even looked at the ones that didn’t require approval, just to make sure that things were working all right. Around about 1997 or 1998, the government decided that it should have a government committee to review what was happening. The House of Reps had a committee review which produced a report called The threat or the glory? Typical spin on this sort of stuff. Again, even though there was no evidence of any hazard or danger, they decided to go ahead and legislate to make all of the regulations enforceable by law. They created the Office of the Gene Technology Regulator and a whole swag of new committees. GMAC now becoming GTTAC but we were originally GTAC (the Gene Technology Advisory Committee). Then some of the activists objected to that and said, no, we were only the technical advisory committee. So we became the Gene Technology Technical Advisory Committee—and the bureaucracy built up.
Yes. It is now very great.
The budget now is $8 million a year or something.
That’s right. In fact, the original basis of concerns about E. coli was because it was a pathogen but it was proved that the laboratory strain was very safe.
It was a worst case scenario. The problem is that we were too clever by far in putting that forward at that time. I don’t think anybody realised how difficult it would be to roll back away from that worst case scenario. As more and more evidence comes out, evidence which says, ‘It doesn’t apply,’ people say, ‘Oh, but.’ Then you get people saying, ‘What about the precautionary principle?’ Nancy told me a funny one about the precautionary principle. What did the bloke say? He took his vitamins in alphabetical order. He said that he didn’t know whether it made any difference, but it certainly didn’t do any harm. That is the precautionary principle.
If you look at it now, there is more than 30 years of experience in work with recombinant DNA in the labs and release. It would really be a great opportunity for some sort of retrospective analysis to say, ‘We’ve approved all this work. How many examples are there of real hazard? How many examples are there where we have avoided a problem by having this?’ They will find lots of examples where they have found people who haven’t been exactly right with the specifications for planned releases – for example the buffers have been 10 metres instead of 15 metres. But that is not what I’m talking about. I’m talking about how many creations will they find where they can say, ‘That was a really nasty thing. It’s a pity. Just as well we had that locked up.’ I don’t believe that there are any. But, on the other hand, the bureaucracy is so well established it would be very difficult to persuade them to do that.
Returning to more personal things, we are sitting in this magnificent mud brick house at Eltham. This is the house that you and Barbie built. I believe that you had mud brick extensions in your first house at Montmorency and now you’ve built this terrific, wonderful house. Do you want to tell us about this?
We had a little wooden bungalow in Montmorency on a block with two big white gums on it, it was lovely. When we came back from the States, we adopted two little girls a year apart. So then in the family there were the three kids and the house became a bit small. Barbie, who knew about these sorts of things much more than I did, got Alistair Knox to design a room on the end of that Montmorency house, and it was lovely. It was typical Knox: slate floor, exposed timbers, big windows and a great big fireplace at the end, and we loved it. We got so turned on by that that we started digging out under the house, making mud bricks. We made a room under the house for Christopher. Christopher made a cubby, down at the corner of the block, out of mud bricks. Interestingly, we went back there the other day, 30 years later. The current occupiers of the house had converted that cubby into an office. So we were all fired up by mud bricks. We even put mud cladding on the timber. We put up chicken wire and mud, so you had fake mud walls.
In 1977, I was in Adelaide attending a course on recombinant DNA technology given by Ken Murray and Noreen Murray. Barbie rang up and said, ‘I have found the block of land that we’re looking for,’ and I said, ‘That’s very interesting.’ She said, ‘There’s only one problem.’ I said, ‘What’s that?’ She said, ‘It’s being auctioned on Saturday’ – and I was getting back on Friday. So I said, ‘Okay. I’ll ring up and have a chat with the bank manager.’ Prior to this, every chat I had had with the bank manager had been remarkably negative. So I was quite surprised when I rang him up and said, ‘We want to buy a block of land, what about borrowing money?’ and he said, ‘Yes, sure, no problems. You can go right ahead.’ The bank agreed to lend us 40 grand. I came back Friday night. We rushed out here and walked around the block and thought, ‘This is really good.’ We came to the auction the next day and I think we were the only bidders, except for someone that the real estate agent had paid to push the price up. Anyway, we bought the block for $42,000 and then we went about selling Montmorency. We sold it for about $50,000, which was a steal.
We got Alistair Knox to design us a house. In the first design the house was up by the fence somewhere. Barbie looked at it and she didn’t like it much. We went and saw Alistair. We were sitting there and Barbie said, ‘I don’t like this, Alistair – dull and boring.’ Alistair looked at it and he said, ‘I don’t either,’ and he ripped it up and put it in the bin. Then he came out, sat on the block and looked at the way the hills went. He decided the only thing to do with this block really was to make a big excavation, cut out this large amount, push it over and build half the house on fill. Then we had a design that we wanted.
Next I got a builder. But, after a week, the builder discovered how little money we had. I said to him, ‘This is the situation. We’ve got 40 grand and I want you to build until that’s all used up and then stop. We’ll finish after that.’ He said to me, ‘No, I can’t do things this way. I’m off.’ Then Alistair arrived with two young blokes and introduced them. He said, ‘This is Tony Ryan and Maurice Wilson.’ Tony was a carpenter and Maurice had been doing medicine. He had got to the part of his medical course where he was involved with patients and had decided that he didn’t want to be a doctor. He took himself to Alistair’s house, knocked on the door and said, ‘I want you to make me into a builder.’ Alistair said, ‘Come and work for me. I’m extending and you can do this sort of stuff.’ Tony Ryan and Maurice Wilson did most of the building, but Alistair persuaded me that I could be the owner builder. I could subcontract and do all sorts of things, including things I wouldn’t normally have thought of doing. We were away.
The first thing we did in the summer was to make the bricks. We had all this soil piled up for making bricks. Barbie and I used to make 100 a day. It was a full day’s work to make 100 bricks. We made about 2,000. I think there are about 3,000 in the house, all in all.
Is it true that PhD students had to make a certain quota of mud bricks before they graduated?
A very interesting observation, Michael. We only had people helping us on one or two occasions and, although those occasions were much more enjoyable socially, we never got any more bricks made. The best day for making bricks was when Barbie and I were just slogging away. The other thing we did was to rush around buying timbers. All the timbers in this place are second-hand. All of these lovely Oregon rafters came out of a picture theatre in Sydney Road that had been burnt down in a fire. They supported all the dress circle. So we got all that. The great big king billy pine posts were bridge timbers. They were covered with bitumen and had nails in them and we had to fix all that up. All the red gums came out of the wharves down at dock 19 where they were pulling them out. During the winter, when we couldn’t make mud bricks, I bought a couple of adzes and we adzed all the big posts. It would take me one weekend to adze each post. We adzed them and we oiled them. We got all the ceiling boards from an old house in Toorak. We took the nails out, turned them over and oiled them. We were incredibly optimistic, Michael. I’m glad that, when we did it, I didn’t really understand what we were doing, because it was a massive task.
What is the grand design thing where they are always going over budget? Yes, we would have been right in there. We didn’t have enough money, in the end, to get the kitchen finished. I bartered what tools I had bought to Maurice. I gave him half the tools I had to finish the kitchen.
We had some pretty funny experiences. We had rented a place in Bonds Road while our house was being built, but of course it wasn’t finished before the rental was up. So we got out of the place we were renting and put a couple of caravans on site. We were living in the caravans. Somebody complained and then the council moved us on. So we moved into the house. We shouldn’t have been in the house because we didn’t have occupancy, but we moved into it. We didn’t have water. We didn’t have electricity. We used to have a fire and boil up a big pot of water to wash the children. One occasion, very Monsier Houlot-like, we were in the tiny little caravan and I was on some government committee and they sent a big limousine with the chauffer to pick me up and take me to the airport. I can still recall when this limousine nosed its way into the building site and I stepped out of the caravan in my suit and tie with my little briefcase heading off to Canberra. But it has been a fantastic experience. It is a wonderful house. It is 30 years old now and it doesn’t look a day old.
It’s proof that the design and functionality have really worked out.
I find this is an interesting question: who have been your scientific heroes, both internationally and in Australia?
I don’t do heroes too well, actually. There have been quite a few people obviously that I admire. In terms of international people, Charlie Yanofsky is someone who has always impressed me enormously with his intellect and his ability to really solve very difficult problems. The whole attenuation thing with trp was an example of that.
Yes. He’s the one who did all of the tryptophan work not only in E. coli in but other bacteria.
That is exactly right. I think he is fantastic. I was very impressed with Doudoroff when I was there. I didn’t know him very well, but he had a sort of intuitive ability. Doudoroff would go to seminars that were on things that were not necessarily his specialty. He had this capacity to cut through with the right question – very clever. Adelberg had what I would regard as a super-analytical mind. He had a mind that was excellent at bringing in facts, organising them in his head and then seeing where the gaps were. It is very different from the intuitive thing. It is a slower process. Obviously, you would have to admire the work from the French school with Jacob and others, they were fantastic. Pritchard in the UK, in the early days, he did a lot of work on the control of plasmids, you really needed to be original to do that. Someone like Sydney Brenner was very impressive, with all of his very clever experiments on r11 mutants of phage.
Yes. I have heard him talk once. But, just from what you read, his ability to express something that isn’t detailed science but is a humorous spin on science clearly indicates how bright he was.
Yes, he was good. So this is the point. You meet people whose intellect impresses you and whose achievements impress you. There is no question: there are lots of people like that in the States and overseas. In Australia, at the top of the list you probably have to have Burnet in his prime as a very original thinker and a great scientist. I think Frank Fenner has to be there for similar sorts of reasons. Then, really, I would put Frank Gibson and Graeme Cox up there too. I was always very impressed with their work and their model on the complex ATPase. It was very innovative and was the first model to come up with rotating subunits. It is a model which is still not so far away from the real situation. I was always very impressed with that and with their ability to do that. Also they did it in E. coli at a time when the general consensus was that E. coli was irrelevant to this problem. They said, ‘No, it’s not.’
Both of those were colleagues of yours in Melbourne and then they both went to Canberra to ANU.
Yes. There are lots of other interesting people, but they are a few that strike me.
Jim, you have had a long career in serving the Australian scientific community by being on granting panels, reviews and also Academy business. Perhaps you would like to compare for us what you think the Australian scientific scene is like now compared to what it has been.
I have been out of it for a bit and it is a bit hard to comment. There is clearly a lot of excellent research being done in Australia. Each year, when you go to the Academy and listen to people who have just been elected or to people who have won the Young Investigator Awards, one cannot be other than highly impressed by the quality of the work that is being done. I think that goes without saying. The nature of the work has changed a lot. Now much more of the work is done in big collaborative teams. The old situation, where you used to have small groups and single people doing research, seems to be on the back-burner.
I guess there is one aspect that I’m not so happy with. It seems to me that there has been an ever-increasing emphasis on outcomes. Although I understand why and I think that scientists should be more active in explaining the outcomes to people. It’s fair enough: if they are getting the money, the scientists have got to show what the public is getting out of it. But it does seem to me that it is becoming harder and harder for people to be supported to do fundamental research. To do research, where all they are doing is trying to solve a conundrum. If you look back in history, you will find that some of the big discoveries that had really important applications came out of exactly this sort of work. The recombinant DNA thing, for a start, is a good example. Herb Boyer and Stan Cohen were not trying to introduce a system for gene cloning. That was not in their minds at all. Herb Boyer was trying to understand restriction. Stan Cohen was looking at antibiotic resistance plasmids and trying to make smaller ones and put them into cells. They both had coffee together and decided that they could join their work and make something new out of it.
I reviewed an ARC application the other day and it seemed to me that the details on the research project over the years have been shrunk in these applications. Initially, that used to be the big thing. Now it is not the big thing. It is there, but there are all sorts of other bureaucratic aspects too. There is one section which really gives me the horrors – that is the section which says, ‘Tell how your research will benefit the nation.’ Really, people write the most ridiculous rubbish. If you are doing fundamental research, you don’t know whether it’s going to work. You don’t know what the outcome will be. The best you can do is say, ‘Here is my track record. This shows you that I’m able to do this sort of work. Here is the importance of this question.’ But I read things where people say that their work is going to save the country and cure cancer, and I think, ‘What are you doing that for? Why are you writing this?’ I know why they’re writing it. There is a list somewhere that says, ‘This section is worth a certain percentage.’
Yes. You have to say how it fits into the national priorities.
In general, I would have to say that I think research in Australia is going very well. Notwithstanding all these problems, there are great research groups and they are making great discoveries. Yes, very exciting times
Yes, I agree that there are, particularly people around about the age of 30 to 40. The talent is still there.
Would you recommend a career in science to young people? How old are your grandchildren now? What stage are they at?
Chris’s youngest, Lily, is doing year 12 next year and she likes maths and physics – I don’t know where she got that from though! Yes, sure I would encourage any young person. I would say, ‘Yes, definitely.’
Why? And what advice would you give to them?
Because it is the most exciting opportunity. We live in a world of extraordinary contradictions at the moment. There is such wonderful new knowledge and new understanding about life coming out of all the scientific discoveries and the potential for all sorts of things just around the corner. Yet we do all this against a background of the most extraordinary radical unthinking pseudo-religious/religious stuff. I would say to people, ‘You need a scientific education and it will be exciting.’ Wonderfully exciting, I think.
Looking back at your life in science and also personally, would you do anything differently?
No, I wouldn’t, Michael. It is the journey really, isn’t it? If you change one bit, that changes everything else. Sometimes the struggle is as important as the achievement. Certainly I think anyone retrospectively might say, ‘Well, actually, if I had thought a bit more about that at that stage, we could have done a slightly different experiment and we might have got there sooner or we might have got a different answer.’ But it doesn’t work that way. You have to deal with it and live with it. Basically, the answer is that I think I have been very lucky. I think it has been a time of enormous excitement and discovery in molecular genetics and molecular biology. I think it has been a privilege to be part of it, and I have really enjoyed it.
Thank you, Jim. This interview has been really interesting and a pleasure.
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