Professor Nick Hoogenraad, biochemist


Nicolaas Johannes (Nick) Hoogenraad was born in The Hague, Holland in 1942. He then spent part of his childhood in Indonesia before immigrating to Australia in 1952. At fifteen, Hoogenraad went to sea but soon returned to finish his secondary schooling at McLeod High School. Hoogenraad graduated from the University of Melbourne with a BAgSc (1965) and a PhD in Biochemistry (1969). During his PhD, Hoogenraad was also a senior demonstrator for agricultural and medical practical classes.

Hoogenraad commenced a postdoctoral fellowship in the Department of Paediatrics at Stanford University in 1970 and was appointed to assistant professor in Human Biology in 1972. Hoogenraad returned to Australia and the newly established Department of Biochemistry at La Trobe University in 1974. Hoogenraad was given a personal chair in Biochemistry (1992), made head of the Department of Biochemistry (1993) and head of the School of Molecular Sciences (1998) – a position he still holds. As well as his research and administrative responsibilities, Hoogenraad continues to take an active interest in science education at both a secondary and tertiary level.

Interviewed by Professor David Vaux 25 November 2010


I am David Vaux and I am interviewing Nick Hoogenraad for the Australian Academy of Science.

An extraordinary childhood

When were you born?

I was born in The Hague, Holland in February 1942. It was the coldest winter on record in an occupied country. It was so cold that you could drive a car a certain distance out from the shore onto the North Sea. My father was fighting in the resistance and my mother couldn’t go to hospital to have me, so I was born in my grandmother’s kitchen. They put cottonwool in the sink and I was born there. In a moment of weakness, my mother said to her father, ‘I’ll call him Nicolaas Johannes,’ which was his name and a family name from way back. I became ‘Nick’ only when I came to Australia and people repeatedly messed up my Christian name.

You mention that your father was in the resistance, but had he trained as something before that?

He was born in Balikpapan in Borneo. His father was an architect who worked for the Shell Oil Company, so he designed a lot of the buildings in Balikpapan. When my father grew up, he came to Holland to go to school and eventually joined the army, with the threatening clouds of the war. He was captured very early – when Holland capitulated. He escaped from the camp that he was taken to and went to work as a civilian in a company in Amsterdam. He was there at the beginning, in 1942, when the Dutch underground started. So my childhood was quite an extraordinary one and I have vivid recollections. Even though I was only four years old when the war ended, I have recollections of being on the run. I remember living in caves and living in farmers’ lodgings, in bedsteads in the wall. I remember sleeping at the feet of a farmer and can remember his filthy feet. I have recollections of sensory things like that. It has left its mark on my own upbringing and family.

What happened after the war?

My father didn’t want to live in a country that had been occupied, because he didn’t know whom he could trust. The Dutch government, in their wisdom, gave a lot of the people from the resistance commissions to go to Indonesia. So he went to Sumatra and became the Commissioner of Police in Medan.

He had already been in that area before?

Yes, he was born in Borneo. He loved Indonesia, passionately. He pined for it all the time that he was in Holland. Eventually, in 1948, we joined my father in Indonesia. For four years we lived in Holland without my father, with my mother looking after the four children.

You had three brothers and sisters. Were you the youngest, the oldest or in between?

There were five of us eventually. I was the second eldest. My brother Robert was 18 months older and we have great recollections together of those early years in Holland and our early years in Australia. Then I have a brother, Paul, who was born during the war. My sister was born the year the war ended and my parents called her Angelique Irene, angel of peace. My youngest brother, Frank, was born in Indonesia.

Your father went to Indonesia and you followed in 1948. How old were you then?

I was six. In 1952, we came to Australia to start a new life because Indonesia had gained independence in the meantime.

Do you still remember much of Indonesia?

Oh, yes. It was a wonderful childhood, carefree and exciting. I remember particularly the last months, sleeping under my bed while bullets came through the roof at night. The freedom fighters were lying in the paddy fields shooting at our house. I remember breaking my arm and going to a hospital and seeing a pile of bloodied uniforms there, where people had been killed.

Who was in charge of Indonesia at that time?

After the war, the Dutch. At the Yalta conference all the colonies were given back to their European masters, which, as we know now, was a dreadful mistake. After 1951, Indonesia became independent.

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A new start in Australia

In 1952 your family moved to Australia. Why did they choose Australia and whereabouts did you go?

My father came straight to Australia in 1951 and he was going to start building a house. We went back to Holland with my mother for a year and then came out to Australia. They made the choice purely to give us a chance at a better education. My father was still not ready to go back to Holland. He was quite traumatised by his experience during the war and suffered continually from nervous breakdowns.

What did your father do when he was in Australia?

He was a clerk. They did not recognise his qualifications in Australia. In 1952, it was a different situation from what it is now. Being an immigrant, even being from Holland, was not a comfortable thing to be.

What was your English like when you first arrived in Australia?

I knew no English when I arrived in Australia. But I was sent to my aunt in Henty in New South Wales, for the first couple of months when I arrived here, and I came back knowing English. In fact, I would say that within two years of coming to Australia, we stopped speaking any Dutch at home. We all learnt it very quickly and we learnt it probably more correctly than a lot of people native English speakers. Those early years in Australia were really difficult years. They were the years that really made me what I am. I went out to do paper rounds at five in the morning from the time I was in primary school to help to support the family.

Whereabouts were you living?

In Rosanna. After doing my morning paper round and coming home and getting changed, I used to walk from Rosanna to McLeod High School. That is where I went to high school, and which was quite a long walk.

School and sport

Tell me about McLeod High School. What was that like?

McLeod High School was only one year old when I went there. In fact, my elder brother was in the foundation year. We had staggered hours. The school wasn’t completed, so we both only went to school for half a day, in shifts. The other half of the day I spent in the Yarra swimming and I became a very good swimmer.

We had very good teachers in those days, particularly English teachers, and they laid the foundation for a great love of the English language. I had a teacher called Ms Hyatt, who was into syntax. We learnt words which, when I use them now, people ask me what they mean. It’s amazing – the education I had in English. We had another teacher who was very good in literature and I fell in love with poetry and things like that. I remember that I used to love the poet Browning. Then I had a teacher in matriculation, year 12, who was terrific at composition and taught us how to write good prose.

Were you very social at school?

In primary school I always hid to have my lunch because I was so ashamed of being sent to school with my sandwiches wrapped in newspaper. So I became a loner and found out very quickly that, to be accepted, you became good at sport. And I did become a sports champion. I was eventually in the Olympic Games training squad.

What was your sport?

When I was at primary school, it was swimming. I went to Heidelberg State School which had a swimming pool. I still have the medal that I won for the Victorian schoolboys’ championship in breaststroke. But then, after the 1956 Olympics, I became really turned on to running and I became one of Australia’s top junior milers. I won the ‘combined high schools mile’, for example.

You were the champion runner at the school when you were in the first year of high school.

Yes. From my first year of high school, because there was only one year higher, I won the school cross-country all six years that I was at high school. I won the district one from middle school onwards basically.

Imagine what would have happened if you had taken up AFL football.

I played football as well. I played all the sports that other people did. I used to go home and do my homework until probably nine o’clock and then I would go for a run for an hour in the dark. I ran with a rock in each hand, supposedly to strengthen my upper body but partly to keep dogs at bay. But, at fifteen years old, I had had enough of this arduous life.

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Run away to sea

What was troubling you? It sounds as though you were doing very well academically and you were one of the best sportsmen in the school. What was troubling you?

I had just had enough of the arduous life I had as a youth. So at fifteen I ran away from home and went to sea.

How did you decide to go to sea? Where did you go to find a ship?

I wagged school one day and went down to the wharf on the Yarra – the wharves were still on the Yarra at that time. I sat there and looked at this gorgeous boat made out of Huon pine, Argonaut II. I remember sitting there, dreaming about what it would be like to go to sea. Then I saw somebody arrive in a fancy car, jump over the gunwale and go in. I thought, ‘He must be the captain.’ So I followed him and asked if he could use a deck hand. He said, yes, he could, ‘We’re sailing at four o’clock. You’ll have to join the union before you go, so you’d better get going.’ So I did. I went home, packed a bag and told my mother that I was going to sea – I am sure she thought I was joking. I went to the union headquarters, joined the union and sailed at four o’clock that night carrying superphosphate to King Island.

How long did you spend on the boat?

I spent three months on the boat. Then, at the end of three months, my ambitions got the better of me. I went to BHP and spoke to the employment officer there to see whether I could get a job as an apprentice deck officer.

You were interested in a bigger boat?

That’s right, and a more professional career. I still remember the man who saw me, Mr Ingram. I was lucky I met this very honest man. He looked at my report card and said, ‘Son, you’re wasting your talents. Go back to school and come and see me again in a year’s time’. Which is exactly what I did.

You went home and just knocked on the door and said, ‘Hi, mum.’

That’s right. I went back to school and then went back to BHP at the end of the year and again he said, ‘You’re wasting your talent cutting your school now; go back and finish matriculation (year 12), and at the end of that we’ll give you a scholarship to do engineering at Melbourne University.’ During my final year at school, I met Joan and I thought that going to sea was not such a good option. I finished up doing what I thought was the next-best option, which was doing agricultural science at Melbourne University. I thought that I would eventually finish up on a research station somewhere in the bush. I thought that would be a great career.

Meeting Joan and a fall-out with religion

Where did you meet Joan?

I met Joan at a youth club and, in a chivalrous manner, I walked her home in the dark. I did that for about 12 months before another guy who would walk home with us said to me, ‘If you don’t grab her hand I will.’ So I thought I had better grab her hand. From there, our relationship developed.

Was this a Christian youth club? Were you religious?

Yes. I went to the Presbyterian Church in Heidelberg and I was religious. But I ceased being religious when I learnt more during my university and PhD studies.

Tell me about that. When did you start to have doubts? When did you stop believing in religion?

Basically, from being fairly flippant at school, and wanting to be accepted, I became fairly serious. Joan tells me that I was always very serious about things. I became very involved with the anti-Vietnam War movement and I started questioning all sorts of things. I used to go to Rationalist Society meetings and I would have philosophical arguments with my PhD supervisor, Frank Hird about free will or the lack of free will. I have had such discussions with you as well, as you know. So out of that came a realisation that there was nothing higher than evolution and natural selection.

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Determined to do biochemistry – and practise it in the vineyard

Who was – how do you say it – Oparin?

Oparin, yes. As an agricultural science student I fell in love with biochemistry. I never did practise agricultural science, except for my own vineyard. In my final year I had to work on a farm and I went to Branxholme in the Western district. I collected a set of books to take away with me and one of them was Oparin’s book Origin of Life on Earth, and from the moment I read that book I was absolutely taken. I wanted to become a biochemist.

Oparin postulated how the first cells might have formed and the first molecules. Then I read more broadly in the area, about these experiments that were done under the sorts of conditions that would have existed on Earth at the dawn of life, and that you could get all the molecules with a discharge of electricity through an environment like that. You could make all the amino acids and even peptides and nucleotides. I was enthralled by that. I have never worked in that field, but it was enough to make me determined to do biochemistry.

You mention that you had a winery. When did you become interested in wine?

When we bought the place we live in now. It is on 25 acres and we had a bit of land that was disturbed because they had put a high­pressure gas pipeline through it. We thought we would plant vines there. This is a good wine growing area, and it was a way of me finally practising agriculture. So we planted 7,000 vines.

PhD with Frank

Tell me about Frank Hird. I remember that he lectured me when I was a medical student.

Yes. He also lectured me and I loved his lectures. But he was a very tough man, as you probably heard. I will just tell you one story that sort of illustrates that. At the end of one year, he took me into his office and said, ‘You can call me Frank now.’ The poor hapless guy in the lab next door heard me calling Frank Hird ‘Frank’ and decided to do the same thing. He was called into Frank Hird’s office and told, ‘I will tell you when you can call me Frank’ – very different days from now. He was a difficult man and very demanding. He would stand over me while I was doing experiments. I used to have a habit of plotting my results on the graph as they came off the equipment and he would watch the results as I plotted them. So I learnt to work under pressure and I think that stood me in good stead. For my PhD, I worked on an agricultural biochemistry problem, working on the role of bacteria to the nutrition of sheep.

Why is that important? What does it do for the sheep?

Sheep eat grass and herbage and it is all fermented in the fermentative organ in sheep. In sheep and cows, it is the rumen, and in horses and rabbits it is the caecum. So I set about trying to find out quantitatively what role the bacteria played. When I was doing that, I got into electron microscopy. I used to go to Frank Gibson’s lab in microbiology to use some of their equipment and I got to know Frank and Graham Cox, and we have remained friends all of our lives. Nancy Millis was my co-supervisor. I discovered bacteriophages in the rumen of sheep, and that was my first paper with Nancy Millis and Frank Hird.

Ian Holmes was on that.

That’s right. I also had a friend from my running days, Bob White, who worked in the University of New England in Armidale. They were doing experiments on sheep that had been surgically modified so that you could measure radioactivity of radiolabelled nutrients administered to sheep by measuring the radioactive carbon in the carbon dioxide in expired air. You could put things into any digestive compartment of the sheep and take blood samples. I made a huge batch of radioactively labelled bacteria based on radioactive wheat that Frank Hird had made by injecting the stalks of wheat plants. He had this wheat sitting in a bag somewhere – can you imagine? So we used that as the substrate for these bacteria to grow in Nancy Millis’s large scale­up facility.

I took my radioactive bacteria to Armidale. Joan and I went in our Volkswagen with our newborn son Andrew and we spent a lot of time there. This ultimately led to a major paper where we discovered that half of the glucose in sheep blood came from the breakdown of bacterial bodies in the normal digestive system. Ron Leng, the person in whose lab I went to work, had been involved with Anison and Lewis in discovering that half of the glucose came from volatile fatty acids, propionic acid mainly. But it wasn’t known where the other half came from. So that is the work I did.

By the end of my PhD I had had enough of working with smelly rumen bacteria. I remember Max Marginson, who worked in Frank Hird’s lab as well, calling me ‘Rumencrud’, as a take-off on my name. I got my own back on him by putting a drop of butyric acid under the armpit of his sports coat, and he stopped calling me that name after that. But I decided, no, I was going to do something different, so I went to work in Stanford.

For the non-scientists, can you tell me what the significance of butyric acid is?

Butyric acid is a volatile fatty acid that sheep make out of cellulose. It is where a lot of their energy comes from. But it is also the smell in vomit and in sweat, and in parmesan cheese, if you want to know.

Rancid butter.

That’s right. I shouldn’t have done that, but I did.

All of the four authors on that paper were really interesting people. What did they go on to do?

Frank Hird’s lab was where just about all the professors in biochemistry in Australia came through in the early days. Bob Symons came through his lab but he had left by the time I had started. Maurie Weidemann, who became professor of biochemistry at ANU and then Barrie Davidson, whom you probably remember also went through that lab. I was there overlapping with Barrie Davidson when Maurie Weidemann was just finishing his PhD. I recall that Frank was in hospital having an operation when he phoned my home. My mother answered the phone and Frank said, ‘Could you ask Nick to come and visit me in hospital?’ I went and visited him and he said to me, ‘I’ve chosen you to be my PhD student.’ Can you imagine that happening today? But I was on cloud nine. I did cartwheels coming out of the hospital. I was so thrilled to be invited to be his PhD student. Frank worked on agricultural biochemistry problems, particularly around this bacterial ruminant nutrition area. He had always fancied having a joint student with Nancy Millis. Because I was going to work on bacterial cell walls, I was it. I am very proud that Nancy Millis, Ian Holmes and Frank are co-authors on my first paper.

Ian Holmes really established the first microscope facility at Melbourne University. He bought a Hitachi and taught me how to do electron microscopy. In fact, out of that, Frank and I made the first atlas of bacteria from the rumen. They were very important days for me.

I remember coming back from Stanford five years later and still being invited to talk about the work I did as a PhD student. Four papers came out of that. But I turned my back on that area of research. I wanted to become a real biochemist and work with proteins, purifying proteins et cetera, and that is what I did.

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Choosing a post-doc

You had to make a choice about where and when to do a postdoc. Can you tell us about that?

Maurie Weidemann, who was a predecessor in the lab, had done a postdoc with Sir Hans Krebs – who was famous for the Krebs cycle. He had left Krebs lab with a very good reputation, Krebs was very pleased to have him. So I contacted Krebs to ask whether he would have me – he was in Great Britain by this time. He said, yes, he would very much like to have me but he couldn’t fit me into his lab for a year, and I didn’t want to wait a year.

Ivan Oliver, who eventually became the professor of biochemistry at UWA, had spent time in Norman Kretchmer’s lab at Stanford University. So, on his recommendation, I contacted Norman Kretchmer and asked whether he had a position for me, and he did. So I went to work with Norman Kretchmer who was head of the paediatric department and they were doing developmental work in mammals.

I had come out of Frank Hird’s lab, where I had had a very strict education. I also had a very Australian education, in the sense that, we still had a workshop just for postgraduate students in biochemistry at Melbourne University and I had learnt to make my own equipment. Frank privately tutored all of his students in how to do glass-blowing and make equipment. Early on in my life, like many Australians, I learnt how to fix my own car. For example I once had to change the clutch in my Volkswagen and things like that. When you went to America in those days you were quite unusual for being handy with your hands.

Those were the very early days of molecular biology and you had to make your own reagents.

The pyrimidine pathway, allosteric enzymes, compartmentalisation and transition state analogs

I didn’t get into molecular biology in those early days (1970). I set about purifying aspartate transcarbamylase, the enzyme of the pyrimidine biosynthetic pathway that was famous for being the regulatory point of the pathway. Aspartate transcarbamylase from E. coli was the classical allosteric enzyme.

What is an allosteric enzyme?

An allosteric enzyme has binding sites that are different from the substrate binding sites, and the binding of a regulatory molecule to this site modifies the conformation of the enzyme and the affinity of the substrate to bind to the enzyme. So you can either positively or negatively regulate the enzyme. All of the theory about allostery really came out of work on haemoglobin and aspartate transcarbamylase. I was going to purify the enzyme from mammalian species, thinking it would be the regulatory point. I had a student colleague who was doing his MD/ PhD degree, Rod Levine, and he had been working on developing a very sensitive radiochemical assay for carbamyl phosphate synthetase, the first enzyme in the pyrimidine pathway. So I decided that, as I purified aspartate transcarbamylase, I would assay my fractions for both enzymes. To my surprise, as I purified aspartate transcarbamylase further and further, carbamyl phosphate synthetase came along for the ride. Rod, Norman Kretchmer and I published that, and it is a highly cited paper because it was the first enzyme complex in the cytosol after fatty acid synthase to be discovered.

It really got me started in my lifelong interest in compartmentalisation, because the first enzyme of that complex was physically connected to the second enzyme. Actually, later it was discovered that three enzymes are part of the complex. I collaborated with George Stark on the complex I had discovered. George was in the biochemistry department at Stanford and was working on aspartate transcarbamylase from bacteria. He eventually took my project further and discovered that the three enzymes were encoded by a single messenger RNA and a single fused gene. There is meaning in this because, in bacteria, the enzymes were single. In yeast, two of them had fused together. And, in mammalian species, three of them had fused together. It was later found that the last two enzymes of the pyrimidine biosynthesis pathway were also part of a complex. That is the theme now. You can have compartmentalisation in the cytosol by having multi-enzyme complexes.

What do you mean by ‘compartmentalisation’?

The first enzyme of the pyrimidine pathway in the cytosol, carbamyl-phosphate synthase, makes the product carbamyl phosphate. There was also a carbamyl-phosphate synthase inside the mitochondria which is a different compartment. Why were those being separated? The mitochondrial enzyme made carbamyl phosphate, serving the urea cycle and arginine biosynthesis. The cytosolic enzyme makes the same product to make the building blocks for DNA and RNA. So they needed to be kept separate. Those pools of carbamyl phosphate had to be kept separate because they provide a different functional role for the cell and therefore they need to be separately regulated. What we also found was that the complex never released its product. So it was not released into the cytosolic soup but was passed straight on to the next enzyme – beautiful compartmentalisation. Then together, Rod Levine and I found that the first enzyme of this complex, carbamyl phosphate synthetase, was the regulatory locus for de novo pyrimidine nucleotide biosynthesis.

So it was different in the mammalian cells from the bacterial cells?

That’s right. I remember that Arthur Kornberg was writing a textbook on Biochemistry and he asked me to write a new section for his new edition of that textbook on regulation of pyrimidine nucleotide biosynthesis in mammalian species.

Who is Arthur Kornberg?

Arthur Kornberg was a very famous biochemist who discovered DNA polymerase and won the Nobel Prize for it. And he was the head of biochemistry at Stanford, absolutely one of the most wonderful departments.

Which department were you in?

I was in paediatrics, but I became friends with George Stark from the biochemistry department because of our interest in the same enzyme in different species and eventually I finished up doing a lot of work with George. He was a chemist by training and he made transition state analog inhibitors. I used those to show that the mammalian enzyme was also inhibited, and it went into anticancer trials to block the supply of the building blocks for RNA and DNA.

What is a ‘transition state analog’?

Enzymes work by taking substrates and, due to a conformational change on the enzyme – which puts stress on these substrate molecules, you get a molecule that is halfway between the substrates and the final products. We call that the transition state. It is the function of enzymes to reduce the activation energy of a reaction so that it will happen more rapidly than it would without an enzyme. George very cleverly designed or predicted what the transition molecule would look like on the enzyme active site and made analogs, which are stable, and they turned out to be superb inhibitors. In fact, eventually, I finished up making a transition state analog inhibitor for ornithine transcarbamylase, the second enzyme in the urea cycle, and in arginine biosynthesis. I finished up putting that onto a resin and developing a single­step purification for the enzyme. Where it used to take a week to purify the enzyme, you could get 100 per cent recovery and 100 per cent purity, in a single step with a transition state inhibitor.

So these analogs could be used to block the enzyme and they could be used to grab on to the enzyme without it being able to let go.

Exactly. We are jumping ahead a bit now, but I became a friend of George Stark. He remained a mentor. In fact, I went back to his lab in the Stanford biochemistry department in 1979 to spend a year with him, and that was one of the most formative years in my life.

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Standford in a vibrant political climate

Norman Kretchmer was a mentor, but also adopted me like a son, partly because of my Dutch background and history. He found out that my father had fought in the resistance and it was important to him because he was Jewish. I felt on top of the world during my postdoc at Stanford. Norman was one of these people who didn’t come into the lab very often. He had very little to do with it. He obtained the grant money. I had come from a very strict background of training and I was very confident. I started lab meetings and I basically became his lab head, helping lots of medical graduates who were coming to do postdocs with their work.

Norman taught me how to write my first research grant. As a PhD student, I didn’t have a clue how research was funded. I didn’t even know about grants. I had never heard of them. I got my first NIH grant with Norman’s help. In fact, after 18 months I was planning to go back to Australia. I had been offered a job in Perth in Western Australia, and Norman Kretchmer said, ‘What do I have to do to keep you here?’ and I said, ‘I want an assistant professor’s position to stay’. So I was made an assistant professor and I stayed for nearly five years.

What was the political climate at the time?

It was vibrant. It was the Nixon years. In fact, after Kent State, where the National Guard turned their guns on the students, pretty well everyone at Stanford Medical School put down their tools and started working to end the war. I joined a small group that went around to high schools talking about the history of the Vietnam War. Norman Shumway put down his tools. Paul Berg actually led a party to Washington to try to see Nixon. Nixon refused to see him. Paul Berg was from the Biochemistry department and he eventually won the Nobel Prize. It was quite amazing. We used to meet at the beginning of every day to plan our day, to see how we could put an end to the war.

Is that where you grew your beard?

I grew my beard earlier than that, when I was still a PhD student and became active in the antiwar movement in Australia. I was well and truly a socialist. I used to go to Communist Party meetings surreptitiously, borrowing Joan’s mother’s car.

What year did you grow your beard and have you ever taken it off?

My daughter is 41 years old and she has never seen me without a beard. I grew it when Joan was pregnant with Kirsten. It has never come off, because Joan said she would divorce me if I took it off.

You mentioned before, at Stanford, Paul Berg and Bob Symons. Can you tell me more about them?

Yes. We came to Stanford, to a new country, with two young children. We had had our two children while I was still a PhD student. Joan joined the International Centre at Stanford and met Verna Symons, Bob Symons’s wife, and we eventually got together. Bob had come out of the same lab where I did my PhD, and from that point we became friends. Bob Symons used to tell me about the exciting stuff he was doing in Paul Berg’s lab. He was on his first sabbatical, as it turned out, from Adelaide University, where he was a staff member. Bob told me about the cloning experiments that they were doing. In fact, Paul Berg eventually won the Nobel Prize for doing the first cloning experiments and Bob Symons was cited in his Nobel address.

I think he was on the key paper on DNA, yes.

The key paper, yes. Certainly, I learnt a great lesson from that. Basically, I had interest across everything. Frank Hird taught me to go to the library and spend half a day there each week, and I read everything. I was always really energised by reading papers. When I went to Stanford, I used to go to clinical-pathological correlation sessions and hear patients discussed. I went to seminars by the world’s greatest scientists and I was very interested in what Bob was telling me.

I went back to Australia in 1974 and in 1979 I returned to Stanford to work with George Stark. I went to work with George with the notion that I could make antibodies against transition state analog inhibitors by attaching them to carrier proteins and make enzymes out of antibodies, ‘abzymes’. That was the first time that was attempted. As it turned out, I failed for reasons I now understand, but other people have certainly succeeded with that.

You were at Stanford University and you had been appointed as an assistant professor. For how long did you stay on?

I stayed another three years as an assistant professor. I was teaching in the Human Biology program. It was a new federally funded program that was established to train pre-med students with no background or very little background in science. I had become involved in teaching in that course as a postdoc. I was a teaching assistant, as so many postdocs are in these universities. My teaching was very highly rated. An alternative handbook was put out by students and they gave me the top rating. I am sure that helped me to get my assistant professor position and probably my position in Australia as well.

Return to Australia

But what happened was that Norman Kretchmer was offered a job to become Director of the NIH Division of Child Health and Human Development and he invited me to come along as his lab head. I did go to Washington DC to look at the lab and I thought, ‘No, it’s not really biochemistry,’ and I was too gung ho to be in mainstream biochemistry working on molecular mechanisms. So I decided not to do that. We got a new head of paediatrics at Stanford who was much more clinically oriented and not so interested in research and I saw the writing on the wall, I would either have to go somewhere else in the States or come back to Australia. Joan was very homesick. I think it is probably fair to say that, if I were not married, I would have stayed in the States. But we came back.

How many kids did you have?

Two children and that was it. They had started school in America. I had an offer of a job at Flinders, which I accepted. Then I also got an offer a bit later at La Trobe, and I knocked back the one at Flinders. I went back to this new biochemistry department, just two years old, at La Trobe University.

In what year was La Trobe established?

The biochemistry department was established in 1972. I went back at the end of 1974.

Who was heading the department?

Bruce Stone was the foundation professor. My return from Stanford was just a total shock. Having come from Stanford into a new department, I was given a lab, which was a large-scale lab, with centrifuges and a scintillation counter in it but there were no benches. I was quite devastated. I felt I had made the worst mistake possible. But, in time, I got things going. I was saved in many ways by David Danks from the Royal Children’s Hospital. He had established a Birth Defects Research Institute and he had tried to recruit me there before I went overseas straight out of my PhD.

What was the initial contact with David Danks?

A letter was waiting for me when I came back from the States asking me to get in contact with him, which I did. He invited me to come to the Children’s Hospital every Friday morning, where we discussed new cases. Coming from Frank Hird’s lab, I was strong on metabolism, and these cases were all metabolic defects. I went there for about three years, spending basically half a day but eventually I couldn’t afford the time as I built my own group. However I remained very close to the Murdoch Institute, as it became known, until David Danks retired. Then later I had connections with it again. In a sense, that activity gave me a way out. As I had credit for my three years in the faculty at Stanford, at the first opportunity I could get to have a sabbatical I went back to Stanford to work this time with George Stark in biochemistry.

You were at La Trobe for four years and then you wanted to go back to Stanford.

Ideas brought back from Stanford

Yes. I had remained in contact with George. There was no email, correspondence was all by snail mail. But I was very interested in the direction in which George was going, having found that the three enzymes that I had been interested in were actually encoded by a single gene. I was interested in George’s chemical approach to biochemistry in making transition state analog inhibitors. I had already made one when I was back in Australia. So I thought, if I could couple my transition state analog, which we called PALO, phosphonoacetyl-l-ornithine, with his, called PALA, phosphonoacetyl-L-aspartic acid, to a carrier protein, so you could inject them into animals and make antibodies, maybe we could make abzymes.

When did you learn the techniques for making monoclonal antibodies?

It was when I went to George’s lab. We decided this would only ever work if we made monoclonal antibodies so that we would have a single antibody with uniform activity. So I started making monoclonal antibodies in 1979. In fact, they were coming out of my ears, I had so many clones.

Did you learn from reading papers –


or were there other people at Stanford doing them?

Not that I was aware of.

So you were making your first monoclonals at Stanford?

Yes, from the Nature paper of Cesar Milstein and George Kohler. Technically, I could turn my hand to things. While I was making monoclonals, I also worked on the western transfer method. It is probably not widely known now that George Stark invented both the northern and the western methods. They appeared in PNAS articles but not as methods papers. The northern was a blotting method that was George’s suggestion and that was taken up by Dave Kemp from Australia, who was working in Hogness’ lab next door to George. The western was another blotting method to blot proteins onto paper and probe with antibodies. But, because the proteins were in cross-linked gels, they were hard to blot. George suggested that I try to develop a piece of equipment where we could pass an electric current at right angles to the way that the proteins migrated in gels, to make them come out of the gel onto paper. I played around with making a number of prototype instruments and eventually got one to work, which I took back to Australia with me in 1980. I had just about everyone around the place coming to my lab to do the first western transfers and also to make monoclonal antibodies, things I learnt at Stanford.

That year at Stanford I got to know Arthur Kornberg better. Arthur was a ‘god’ professor of the old-fashioned sort. When he started the biochemistry department at Stanford, he brought across with him his department from the Washington University in St Louis, Missouri – people like Paul Berg, Dave Hogness and Dale Kaiser. He insisted that they have lunch with him every day. People would bring in their nosebags, eat lunch around a table and talk about science ideas. When I went there in 1979, I was invited to these meetings. It was called the ‘Wednesday Club’ by this time, because they only met on Wednesdays. Everyone was too busy to meet every day. I was afraid to open my mouth for the first month or so. But, after that, you learnt that your ideas were as good as anyone else’s. It was this fantastic reinforcing, by all these really world-class people. No­one could fail in an environment like that.

George came from Moore and Stein’s lab, famous for inventing amino acid analysis. George started as a chemist but he eventually became one of the world’s great molecular biologists. He discovered the Jak-Stat pathway, and, with the help of postdocs who were there, he also developed the northern and western methods. George has been inspirational to me, as was Arthur. The ‘Wednesday Club’ was an example of what to do, and I set up something similar in Australia when I came back.

How important do you think new technological advancements in science are?

Absolutely essential. It was all about making things. The scientists had to make the first instruments. But it has always amazed me how quickly companies come up with first class commercial equipment such as protein sequencing machines. The first mitochondrial enzyme I purified was ornithine transcarbamylase, using my transition state analog inhibitor which I covalently attached to an affinity support. But we were also the first to clone a cDNA for a mitochondrial enzyme from a mammalian species. This was made possible because I learnt how to do protein sequencing manually using the Edman method, and I learnt how to make DNA manually using a Pasteur pipette and test tube.

Where did you clone the gene?

Back in Australia. I learnt the basic techniques for cloning from another thing that Arthur insisted on. That was that each of his staff could only have an office and a small lab, all the other researchers were mixed together in other labs. I was working with George Stark, but I wasn’t working in his lab. I was working in a lab with another PhD student of Dave Hogness, Jeremy Nathans, who was making DNA manually, synthesising small length of DNA called oligonucleotides. I took note of all this and wrote notes so that, when I came back to Australia, I could do exactly the same thing. It is very powerful to put people with different research interests and expertise in the same labs because you encourage cross-fertilisation. It is an example I have tried to emulate since then. Arthur also created an environment with enormous discipline. I never heard anyone stab anyone else in the back or speak negatively about their colleagues, even though, in a department like that, with two Nobel laureates and others who might well have won the Nobel Prize, there was a lot of tension. But it was just the most wonderful place to work.

You were there for a year and then you returned to La Trobe all fired up.

All fired up and didn’t look back. As I said, I started my own equivalent to the ‘Wednesday Club’, where I went to other biological science departments. I remember that Jenny Graves, George Stephenson and various other people used to come. We would take it in turn to talk about ideas and about the research that we were doing. I also came back with an apparatus that put me in charge of westerns. Robin Anders, whom I knew from PhD days, came from WEHI to use my instrument. Ian Mackenzie also came to my lab and learnt how to do westerns and they copied my instrument.

You brought the technique for western blots and for making monoclonal antibodies –

Other people had done that as well. People in New South Wales were making monoclonal antibodies. But I was certainly one of the first ones.

And for synthesising oligonucleotides.

Yes. There was a team doing that at the Howard Florey as well and Ian Mackenzie was making monoclonals probably earlier than I was. All these methods came in from overseas.

Then they all got fertilised all around Australia.

Then I had a wonderful PhD student who came in and said that he wanted to work on molecular biology. That was Peter McIntyre, who is now professor of Pharmacology at Melbourne University. I put Peter on to cloning ornithine transcarbamylase and carbamylphosphate synthetase, urea cycle enzymes. In those days everything was manual. It took all night to make a 15-mer with 64-fold complexity with just a Pasteur pipette and glass beads on a sintered glass funnel. When we got our first cDNA clones, you had to make the libraries yourself. They were made by Julian Mercer and Peter Hudson, they collaborated and made the libraries. We screened them with oligonucleotides and then manually sequenced them. These days we have machines for all of those things.

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Crowded mitochondria

You were interested in the enzymes that were originally found in bacteria but then found in mammalian cells.

Yes. Mitochondria evolved from alpha-proteobacteria from a bacterial symbiont. The cell before that, the archaeal cell, was an anaerobic cell. They didn’t have the mechanism to utilise oxygen as an electron acceptor to make energy in the form of ATP. By setting up a symbiotic relationship with this alpha-proteobacteria, they were able to become oxidative. Therefore, in a sense, it was an important process in leaving the sea and making very efficient use of energy, because the oxidative metabolism is a very efficient form of making energy.

But my interest in the mitochondria, in a sense, came from my interest in the compartmentalisation of metabolites. I remember in the early eighties reading a paper by Daniel Atkinson from UCLA, who spoke about the solvent crisis that cells have. This really resonated with me. There are so many solutes in the cell – proteins, nucleic acids – that the cell is right on the edge of a solvent crisis.

You mean turning into a solid.

Yes, a solubility crisis. We know only too well that, if you make too much uric acid, it comes out as solution and produces gout and kidney damage. We know now, much later, that many of the diseases of old age are all about proteins coming out of solution. It became clear around this time in the early eighties, when people started doing calculations, just how extraordinarily crowded the cell was. Mitochondria from liver cells have around 500 milligrams per ml of protein in them. It is just unbelievable. When we were purifying enzymes, if we could get 10 milligrams per ml, that was great. If you tried to go higher, it would come out as a solution. But, in a cell, you have 500 milligrams per ml in the mitochondria and in the cytosol, 250 milligrams per ml.

I have heard that in some protein crystals the concentration of protein is less than 5,000 milligrams per ml.

The question of how the cell can cope with such a crowded environment is a huge puzzle and I really think that is something for the future. The huge puzzle is that, despite the whole cell having this crowded environment, things can diffuse freely. You can use NMR to show that there is free diffusion of protons within the mitochondria. Yet, if you do the calculations, there is only room for two water molecules around each protein molecule at 500 milligrams per ml. So it probably suggests that the organisation of proteins in the cell is even more highly structured than we believed. Probably there is a soft structure in the cell that is immediately destroyed when we break open the cell. It is very difficult to work on this. We need new methods to try to find out how this works. It is probably organised in a way where metabolites are passed logically down pathways.

How many different proteins are in the mitochondria?

Altogether, there are probably around 1,500. But it varies in different tissues. There are enzymes present in mitochondria in some tissue which other tissues do not have. Like the liver and the small intestine which alone have the urea cycle enzymes in mitochondria. So, on average, maybe there are 1,000 different types of proteins in mitochondria in most tissues.

Where are those proteins made?

All but thirteen of them are made, or encoded, in the nucleus. The poor old mitochondria has held on to just a bit of DNA so they can make a dozen proteins. They need all the machinery for making proteins, but nearly all of the proteins come from the outside. The genes for those proteins have been moved, over evolutionary time, into the nucleus.

Can you tell me how the mitochondria and the nucleus talk to each other?

Yes. Again there are always these seminal papers that really change the way that you look at things. There was a paper in 1989 from Eilers and Schatz and there was also a paper from Walter Neupert’s lab, which suggested that proteins had to be unfolded to get into mitochondria. Jeff Schatz was in Basel and Walter Neupert was in Munich. In fact, the way Schatz did it was by putting a signal peptide for getting a protein targeted into mitochondria onto an enzyme dihydrofolate reductase. If you add the very tight binding drug methotrexate, which is really a transition state analog inhibitor, to that enzyme, it folds very tightly. It is knotted, if you like. As a result, that protein now will not be imported. I remember reading Nature News and Views in which Roger Kornberg and Jim Rothman wrote an article predicting that people would find an unfoldase on the surface of mitochondria that is responsible for unfolding the protein before it gets in.

As it turns out, that was an incorrect prediction. What was found to be responsible for keeping proteins unfolded was that proteins remain unfolded from the time that they leave the ribosome until the time that they get to the mitochondria, due to interacting with this new class of molecules called ‘molecular chaperones’. A beautifully apt name because they stop the protein from making unwanted liaisons with other proteins. But they are not part of the final product. They do this catalytically. Some of them have enzyme-like activity. That is, they change shape back and forth.

Protein import to the mitochondria

My lab immediately became interested in the role of chaperones in protein import. Proteins destined for the mitochondria have an address signal on them and the matrix proteins have an extension on their N- terminal end. In the ornithine transcarbamylase case, it was 32 amino acids long. We became interested in that because, when we cloned ornithine transcarbamylase, we discovered that there were 32 amino acids in the sequence that I hadn’t found when I had done the N-terminal sequence of the mature purified protein. So we became interested in what those 32 amino acids were doing.

There were 32 amino acids that had been removed somehow.

It was pretty easy to make the prediction that they were there to specify where the postman should deliver the letter, the address if you like. A series of PhD students in my lab looked at that particular question. We did it in mammalian species. Schatz and Neupert were the main competitors, Schatz in yeast and Neupert in Neurospora. I guess our progress was fairly slow because we were using mammalian species and we didn’t have the power of genetics on our side.

But I finished up joining forces with a Japanese person called Masa Mori, who became a close friend. I found that he was almost my mirror image in another country with a love of classical music and a love of woodwork, and he was choosing exactly the same questions to answer as I was. It became disconcerting and I wrote to him and said, ‘I think we should talk.’ We did talk, and we met and collaborated from that point on, which was lovely. That is one of the wonderful things about science.

So these proteins are either delivered to the mitochondria or they find their way there. There is machinery in the mitochondria that proofreads the signal and says, ‘Yes, you can come in.’ Then the precursor protein gets pulled in by a machinery inside the matrix of the mitochondria. The machinery is now well understood. It is just like machinery – like tugging on a piece of string and pulling it in. Then, when these proteins get inside, the signal peptide is cleaved off so that it can never leave again. It is now trapped inside. It was later found that proteins that are destined for the outer and the inner membrane, proteins of the electron transport chain and so forth, have a different sort of signal. They have an internal signal which is not cleaved off when they get in. They become embedded in the membrane and it keeps them there.

In fact, just to close the loop, I became friends with Ulrich Hartl, who is the director of the Max Planck Institute of Biochemistry in Martinsried, just outside of Munich. We were both invited to speak at a conference in San Antonio. The organisers left us standing alone there at the end of the talk, so we said, ‘Let’s go and have a meal together.’ From that point on, we became friends. I think he is probably one of my closest friends in science now. As I became more engaged in administration, growing a department and a school, Ulrich threw me a lifeline by inviting me to come out to Munich to look at something where he thought our fields overlapped. He works on molecular chaperones. He is one of the world’s top workers in that field. I went there to work and spent a week floundering around and then found my feet again. By the time I had left at the end of six weeks, I had made a very important discovery that HSP90 was involved in protein import.

It was one of those chaperones?

It was one of the chaperones. It hadn’t been suspected, because it wasn’t involved in yeast and Neurospora, so it had been missed by the people making the most progress in the field. I took a three­month sabbatical the next year and went back again and found that there was a very large complex. It was like a space shuttle that shuttled the protein from the ribosome, in an unfolded confirmation, to dock with a receptor on the mitochondria called TOM70. It then handed over the protein. The docking process caused the chaperone ATPase activity to trigger, causing a conformational change, releasing the cargo so that it could now go into the channel.

Into the mitochondria.

We found by looking again at old work that, in fact, most proteins that go to the matrix don’t need chaperones, despite the claim in the literature and in the textbooks that they do. They don’t need chaperones as they keep out of harm’s way by folding.

In the cytosol?

Yes. When they get to the mitochondria, their signal peptide pokes into the mitochondria and it is grabbed by a chaperone that works like a ratchet at pulling it in. It unzips the folded protein, making it become unfolded and then the protein refolds inside. The strength of the folding of a protein is very, very small. It is only about 25-kilojoules per mol, equivalent to just a few weak bonds. Proteins are very fragile because they need to be fragile to work. Conformational change is what makes proteins work. But when proteins are folded it stops them from aggregating and getting into trouble.

We published our work in Cell together. I kept going back to finish this work and it took a long time to finish it. But it was my haven away from administration to be able to go back and work with my own hands. A postdoc, Jason Young, finished up joining in after the early work that I had started and he really saw the thing to completion. So Martinsried has been a special place for me to go to and get relief. The model now is that proteins are either folded and then unfolded by this mitochondrial machinery, or they are retained in an unfolded state by the large cytosolic shuttle complex and then get inserted into membranes or even into the matrix.

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Mitochondrial stress response

You have also done some exciting work on the mitochondrial stress response.

Yes. Where that began was that I started this collaboration with Peter Hoj. He joined the department as a postdoc working on plant things with Bruce Stone but he wanted to work on the mitochondrial import problem with me. So we started emphasising the isolation and characterisation of all the molecular chaperones that were found in the mitochondria, Peter led a lot of this work. Interestingly enough, the molecular chaperones looked very much like their bacterial counterparts. You could see that they had evolved from bacteria because they were different from the mammalian ones that were outside the mitochondria. Collecting things like that is a bit like collecting postage stamps. Somewhere along the line, a New Zealand postdoc called Ryan Martinez came along and we decided that we would work on the function of the mitochondrial chaperones. What were these chaperones actually doing? So Ryan made some rho zero cells. That is, he made cells where we removed the mitochondrial DNA out of the mitochondria. You can do this chemically with ethidium bromide.

He made these and we looked at what happened to the chaperones. To our surprise, we found that the chaperones of the mitochondrial compartment were upregulated and the chaperones outside the mitochondria were unaffected. Chaperones, I should point out, are regulated by stress. When a cell is under stress, proteins become unfolded because they are soft and they have very weak forces holding them together. To salvage the cell, to stop it from dying, the genes coding for chaperones are activated. You get more chaperones and that helps to save the cell. If that doesn’t work, you get proteases induced and, basically, the unfolded proteins are removed from the cell to clean up the mess. The final step, if that doesn’t work, is that the cell undergoes apoptosis – programmed cell death. It is better to get rid of a non-functional cell than to leave it there.

The mitochondria also need to have chaperones. Where are the genes for the mitochondrial chaperones?

They are also in the nucleus. The genes in the mitochondria only code for 13 polypeptides of the electron transport chain. They are all membrane proteins. Presumably, the proteins encoded by those genes are so complex that it was difficult to transfer their genes to the nucleus and to get the proteins in from the outside. At that point we discovered a stress response pathway. If you heat cells, you increase the vibrational activity or energy in the cell and proteins become unfolded. You induce all these chaperones and proteases as well. But this wasn’t what was happening in these rho zero cells. The cytosolic chaperones were unaffected and only the mitochondrial chaperones were upregulated. So we discovered what we call the ‘mitochondrial stress response’.

From there, I had a Chinese PhD student, Quan Zhao, who followed this right through and found the promoter regions of the chaperone gene through which this particular activation worked and the transcription factors that worked through this. The mitochondria sense the presence of unfolded proteins. It then signals to the cytosol to start a cascade of protein kinases being phosphorylated and activated until transcription factors are induced, and we found that part of the pathway. You make the transcription factor, and this in turn activates a large suite of genes that save the cell from the disaster of having a non-functional mitochondria.

The part we haven’t discovered yet but which Ulrich and I are talking about ways of doing together is the sensing. We know that there are proteases induced in the mitochondria as a result of unfolded proteins accumulating in the mitochondria. These proteases clean up the mess, so the stress response is quite reversible. That means that there has to be an increased flux of peptides out of the mitochondria, and we think that this may be the signal. The hypothesis that we need to test is that there is something in the cytosol that is sensing the flux of peptides.

And the signal goes from the cytosol to the nucleus?

Yes. I think it has been very exciting to find a new biological process in mammalian species. We wanted to be able to use a genetic approach to try to define all the steps in the process. Quan spent more than six months trying to get it to work in yeast and we never found it there. In fact, we then found that somebody in Walter Neupert’s lab in Munich had spent a year on trying to find it in yeast and couldn’t find it. But, more recently, David Ron has found the same process in an invertebrate species. I gave a seminar on the mitochondrial stress response in New York and David Ron became interested in it and turned to C. elegans to study the mitochondrial stress response. He has found this pathway in C. elegans and he can use a genetic approach. But, interestingly, in C. elegans, it is very different from the mammalian pathway. There are different transcription factors and so forth. Also, the stress response pathway – the ‘mitochondrial unfolded protein response’, which we now call it – is only present in developing C. elegans but not in mature C. elegans.

How do you specifically cause stress just in the mitochondria?

The way we did it was by taking this favourite old enzyme of ours, ornithine transcarbamylase, and knocking a piece out of the middle. This piece is where one of the substrates, carbamyl phosphate, binds. We theorised that this piece was exposed to solvent and so, if we took it out, we would prevent the protein from folding correctly. That is exactly what happened. It was still targeted to the mitochondria, because it turns out that ornithine transcarbamylase doesn’t use the pathway where a protein folds and is unzipped. It uses the other pathway with this large complex with HSP70 and HSP90. Once the protein goes into the matrix the signal peptide is removed. We found that this mutant form of ornithine transcarbamylase got into the mitochondria just as efficiently as the wild type enzyme and it was processed. But it couldn’t assemble into a trimer and it couldn’t fold properly.

I am interested in the way that things occurred. Did you make the mutant protein so that you could study the stress response, or did you discover the stress response and then want to figure out how it worked?

We made it deliberately to study the response pathway because Ryan Martinez had decided to make rho zero cells. Rho zero cells have their DNA knocked out of them. So they can’t code and thus they can’t make those thirteen polypeptides that are needed in an electron transport chain. The complexes of an electron transport chain are massive. There are more than forty proteins in complex one, for example. If you can’t make some of those subunits, then the ones that are imported from the outside to be assembled with the complex, can’t assemble. The theory we had was that it was a way of causing the mitochondria to accumulate unfolded proteins. And, sure enough, that is exactly what happened. And this enabled us to discover this mitochondrial unfolded protein response.

We deliberately went looking for it because there was an endoplasmic reticulum specific unfolded protein response and we wondered whether there was mitochondrial one as well. We found it with the rho zero cells, but rho zero cells were awkward to work with, because we couldn’t reverse it. We couldn’t manipulate it. Once you knock the DNA out, you can’t put it back in there. You have got no template to make it in the mitochondria. So we looked for another method of making unfolded proteins accumulate. We did it by making a mutant form of ornithine transcarbamoylase. We made several mutants and they all worked, but the deletion mutant was the most convenient one.

I am interested in the impact of new techniques. Brian Seed came up with a novel way of cloning genes for receptors and you have used this technique. Can you tell me about it?

Yes. I spent a year’s sabbatical in London at the Imperial Cancer Research Fund (ICRF), as it was called then, working with George Stark and Ian Kerr. George had moved into the field of Interferon. It was 1988 and it was an important sabbatical for me because it really got me into cell biology. It was a prelude to me being able to do the work we were just discussing, with the mitochondrial unfolded protein response. Towards the end of my stay in London I was visiting a lab at the Radcliffe Hospital, just outside of Oxford, and talking about my work. And, as part of trying to find a way to clone the Interferon receptor, I mentioned that I was probably going to next try to use Brian Seed’s method to try to get the Interferon receptor.

Brian Seed had developed many methods. He was one of these crazy scientists who lived at work. You would go into his lab and he would have huge stacks of empty coke cans around the place. One of the methods that he came up with was called the panning method, whereby he developed a very special vector. It had lots of really great innovations in it. It was very small so that you could put large cDNAs in it, such as you might expect to get for receptors. When you transfected that vector containing a library of cDNAs into cells, you get expression of the receptor. So, when you put antibodies on the bottom of a petri dish it would bind to those cells which expressed the receptor that the antibody was against. You could wash it – amazingly, it is like holding a human being by a single hair, basically holding a cell by an interaction between an antibody on a plate and a receptor in the cell surface. That was his method and I was going to use it.

I was given a Brian Seed library from foetal liver. I took it back to Australia and I duly put it in the freezer, waiting to use it. One day Nick Gough from the Walter and Eliza Hall Institute gave a seminar on his work on cloning receptors involved in blood cell formation. He told us about the extraordinarily difficult task of getting enough protein purified, to be able to get sequence, so that you could make oligonucleotide and go through the routine cloning methods. During drinks after the seminar, I said to Nick, ‘Have you considered using the Seed method?’ which he didn’t know about. A lot of Seed’s stuff wasn’t published necessarily. It went around by word of mouth. I said, ‘If you are interested, I happen to have a library, you’d be welcome to use it.’ Duly, Dave Gearing came and took aliquots of the library and all the rest is history. As you know, a lot of receptors were cloned at WEHI using the library that I brought back with me from England.

Democratic department

Let’s switch back to La Trobe University and the person who recruited you there. Can you tell me some more about Bruce Stone?

I knew Bruce from my PhD days. He was at Melbourne University and worked on the same floor that I worked on. He was in the lab opposite. I knew him from social functions and other things. One of the reasons for my deciding to go back to La Trobe was that I respected Bruce and I thought he would run a good department. Initially, my first period before I went back to Stanford was pretty ordinary. I had to prepare new lectures and I didn’t have a lab really. I have been very conscious ever since then never to recruit a person without being ready for that person to come and have facilities available. Certainly, there were no facilities available for me.

Bruce was also of the old generation. I remember one day reading a newspaper ad for a new lecturer in the biochemistry department at La Trobe and being quite upset that I had heard no mention of this, no consultation. I was, I guess, a bit of a bolshie because I felt that we were all doing our best to make the department a good department and we were working very hard and conscientiously and introducing lots of new techniques. Therefore, it was our department as much as it was his department. So I got my colleagues together and we agreed that we would take him out to lunch. We took him out to lunch and told him about our dissatisfaction, and he said that he would try to be more consultative in future. These are lessons you learn and you store away. The whole principle of having co-ownership with the people that work in your place is really important. People work harder for something they own rather than something that somebody else owns. In any case, it is the democratic thing to do, so it seems right to me.

Certainly, the department changed and I think Bruce built a very good department. It is interesting because this question keeps coming up from people but I think the reason for that comes from this: do you recruit into particular areas that the department is already strong in or well known for, or do you recruit the best, irrespective of what the area is? The latter is exactly what we did. When a position was advertised, the best person got the job. It is a policy that I notice Paul Nurse is now going to use in developing his new institute in London. It is a pretty flawless way to go ahead.

Through your experiences, you have developed strong opinions about how a department should be run. Can you tell us about those and how you are going to use them at La Trobe in the future?

Yes. It is a question of finding the best people and appointing the best people. What has happened in more recent times – and ‘recent times’ are perhaps the wrong words, because I took over from Bruce Stone in 1993. I was head of the department while Bruce was still there. Towards the end, before his retirement at 1993, Bruce handed over the head of department to a number of senior members of staff and I had a couple of terms, of a couple of years, being head of department. I got a personal chair before Bruce retired. So, when his position became vacant, I applied for it as much as a blocking bid. If they couldn’t find anyone better than me, they would be better to stick with the devil they knew than to take a risk. I applied and I got the position, and I have been in that position ever since. I have probably been the longest serving head of department currently in Australia, although Mike Ryan has recently taken over as head of department.

When the university established schools and I became responsible for the departments of Genetics and Chemistry – and eventually Pharmacy in Bendigo as well – the job became a very heavy one. But I was determined to build a department based on recruiting people who were looking to start up their own labs. That is, people who had fellowships so that they were self-funded, but who were very keen to break away from a group that they might have been part of and start their own lab. I think that has worked really well. As you know, we have expanded the department substantially with the sorts of people who have been in a strongly mentoring environment. Not just mentoring by me, but other people who have walked the same journey and who can help them with applying for grants and managing budgets and students, things that don’t come automatically to some people.

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Supporting secondary education

I know that you recruited Francesca Calati. Can you tell us about her role?

My philosophy about education is that we should vertically integrate the education process. What I mean by that is within the same environment, we should have not just researchers and post-graduate students but also the best undergraduate students. That way they can start to get a feeling for what it is like to work in a research lab in a research environment. Also, we have an obligation to help with the education of kids before they come to university. In universities we are only too ready to complain about the quality of students we get in, and to complain about the curriculum that students are taught, but we have rarely seen it as our responsibility to make a contribution.

I have a son, Andrew, who is a teacher. I know from him and his colleagues how dedicated they are, so I am not particularly happy with complaints about the secondary school education system that we tend to have. When he started teaching, he trained as a zoologist but couldn’t get a job in ecology at the time – this was at the time when Kennett was premier and laying off lots of teachers. So Andrew retrained as a physics teacher. He went to Tallangatta up by the Hume Reservoir to do his first teaching because he was mad about outdoor activities. He had been brought up to ski and rock climb, so he went up there. I discovered, to my horror, that that these kids in Tallagatta secondary college were educationally underpriveledged. Some of the children in his classes would have milked cows before getting on the bus for half an hour or an hour to come to school. They were exhausted or tired and couldn’t learn properly. They even went so far in their VCE year, the final year, to pool money together to put somebody on the train to come to Melbourne to go to a workshop and then pass all the notes around.

I felt we needed do something about it. We had a little bit of money from full­fee income and I put it to the staff that maybe we should try to hire a teacher to make a CAT website. In years 11 and 12, they had Common Assessment Tasks (CAT) and students had to do projects, and I thought we had the sort of project material that students would love to have. I went over to the Victorian Science Teachers’ Association conference to try to find somebody to put out an announcement or a notice. I had things written on a bit of paper and I met this person and asked where the president was. She said the president wasn’t there, but I could give the bit of paper to her. She shoved it in her pocket and I thought, ‘That’s the end of it.’ But she phoned me. Her name was Jenny Herrington and she was the chief examiner in biology and Vice President of the Science Teachers’ Association of Victoria. Jenny phoned me and said, ‘I’m the person you’re looking for.’ She got a secondment from Caulfield Grammar for a year and we paid her salary. She stayed on her school salary, but we reimbursed the school to find a replacement. She interviewed all of us and put project material on the website, which we call the ‘CAT site’.

It absolutely moved the world. I couldn’t believe what a resonance it had. She had constant emails from schoolteachers in distance schools that didn’t have that sort of information. She knew exactly what the examiners were looking for in biology. To help them, she put her notes on the website. Also, lots of schoolkids came into the department. We gave her an office and it was always full of people. She won the BHP Science Teacher of the Year Award.

What happened to this website?

Eventually, we had to pull it offline, because a website that you don’t keep up does more harm than good, and we simply didn’t have the money to do it again. But we stored this away as an exercise that had been very worthwhile and we wished that somebody else would have taken it up for the small amount of resources.

Andrew eventually became a teacher in Ivanhoe Grammar School, which is closer to the university. One day over a family dinner, probably after a couple of wines, he suggested to me the notion that fifteen­year­olds in year nine, are at the bottom of the educational profile. He said many schools had special programs for these kids. They are going through their body changes and they think they are adults when they are not. We thought about having a program of bringing those students to La Trobe University to be given a lecture by people like you, me et cetera and they could then work in groups to do project work. This was an experiment that we did with Ivanhoe Grammar School. It turned out to be extremely successful. Ivanhoe Grammar School was very positive about it and wanted it to continue.

We then found the money to advertise for a person and, to my absolute delight, Francesca Calati applied. She had just won the Prime Minister’s prize for teaching. She had been at St Helena Secondary College, a state school, where she had developed a nanotechnology program to lure kids into physics and chemistry. It was so successful that she developed an accelerated learning program. In fact, the nanotechnology program has now been adopted by the government and has been put out throughout Australia. Francesca has been involved with that while working with us. She has expanded the program. The students get office space in the corner of the department and learn how to make appointments with busy people. They come in with their little tape recorders and typed questions and ask whether they can tape your answers. It is just delightful to me. It is just great.

As part of this vertical integration, you have already mentioned students in year nine. Can you tell us about other aspects of this process?

Yes. Francesca is expanding the program to have students also coming in at other levels. Also, she has developed some ideas about getting their teachers to come in to bring them up to date because technology is changing so quickly. We are going to be faced with personalised medicine, before people are ready for it and before the community knows about it. It is really important to educate and to bring the educational program further. In the recent international conference that we organised, I asked you to organise a free public forum for the public to come in and be part of the conference and to hear what is going on. In this particular case, it was in the environmental or alternative fuel field, but it could be in any field. I think we need to try harder to involve the public in what we do.

You have told us of your work concerning very basic biological processes, such as transport into and out of the mitochondria. Do you think any of this is going to have any practical or medical benefits?

Of course, we like to get funded by NHMRC, so we have to make a medical story. For all the work we do, it doesn’t matter how basic it is. Of course, I genuinely hope that a fundamental homeostatic mechanism, such as the mitochondrial unfolded protein response, has medical implications. So I, or people in my lab, have spent a lot of time handing out clones and all the reagents for studying the mitochondrial unfolded protein response to people around the world. This has struck gold, so to speak, with a group in the Technical University of Munich. It has taken a long time, but now this group has confirmed all of the discoveries we made, but taken it a step further and found that the mitochondrial unfolded protein response is somehow implicated in inflammatory bowel diseases, such as Crohn’s disease. So, of course, I was absolutely delighted that this connection has been made. I am sure there are other connections to be made because I have certainly given all my reagents and clones to people working on liver disease and various other things like that.

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Work/life balance?

Your wife, Joan, didn’t train as a scientist, but she certainly spent a lot of time in your lab. Can you tell me how this came about and what it has been like?

I told you earlier about making monoclonal antibodies in 1979 in Stanford because we were trying to make antibodies with enzyme activities. I got to a point where we had literally hundreds of clones we were following up and I was just overwhelmed by it. Joan used to pick the kids up from school and come into the lab and they would sit endlessly and wait for me to get ready to come home. So I went to George Stark and said, ‘What would you think, George, if Joan came in and helped me to do the tissue culture to make monoclonal antibodies or to look after them?’ I always regarded tissue culture as a bit like gardening or cooking, you just have to be careful about what you do and you need to have an eye for things. He said, ‘That’s a wonderful idea. Mary’, his wife, ‘used to work with me once and maybe she’ll come in and work as well.’ In fact, both of them have worked in the lab since then. But now George has now retired and so has Mary. Joan came in the lab and eventually took over the tissue culture facility and has made monoclonal antibodies with lots of people and for lots of people. She is now involved with training the master biotech/bioinformatics program students. So, yes, she has worked in my lab and as part of my lab, but for quite a long time she has been working not with my lab so much as with other people. It worked well for me, because you know what scientists are like, their jobs are very demanding. We work long hours and to have your partner in the game makes them more understanding at least, and I think that is enormously helpful.

And you have four kids?

Two children. Andrew, the teacher, I just mentioned, and Kirsten, who is a clinical neuropsychologist.

And grandchildren?

Yes, five grandchildren. It is a great time in my life.

Tertiary teaching

What do you see as the importance of teaching in university and research institutions?

It is interesting because, as you have heard from my story, I have been very influenced by what I learnt in America. One of the things I learnt in American universities was that very active research scientists also gave lectures. It was just part of their life, basically. Paul Berg used to give his semester-long lecture series on oncogenic viruses and people were hanging from the ceilings to hear him talk because he is such a wonderful lecturer. I learned from his lectures the importance of being able to get across a wide range of audiences. He had students who had never heard any this stuff and he had postdocs who were working in the area and they were all enthralled by what he had to tell them. He had this wonderful ability to go across the broad disciplines. I recently heard Harvey Lodish give a talk. He has written the top textbook in Biochemistry and Cell Biology. He is at the Whitehead Institute. He said that every person in the Whitehead Institute, none of whom are paid by MIT, gives undergraduate lectures.

I regret that in Australia there is too much segregation of people who do research only and those who do research and teaching. I think it is unnecessary. People who have research-only positions feel afraid to take time to give lectures, because they think they won’t be competitive for grants. But, somehow or another, this fear isn’t shared by people in America. So I would like to see more contributions from people who have a hell of a lot to give. I can imagine first­year students being given lectures by somebody who is a really famous or good scientist being forever turned on by that experience, and we need these people to go into science.

Thank you very much, Nick, for talking to us today.

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