It was with great sadness that we learned of the death of our friend and colleague Richard Jackson, who passed away peacefully on Monday 21st September.
Richard had a long and illustrious affiliation with the Department of Biochemistry. He graduated 1st class from Part II Biochemistry in 1962, and then carried out his PhD with Asher Korner, alongside fellow PhD students Tim Hunt and Tony Hunter, producing a hefty two-volume thesis on polysomes. After a postdoctoral position in Geneva with Alfred Tissières, he re-joined the Department for the rest of his career. Richard formally retired as Professor of RNA Biochemistry in 2007; but he never really left, continuing until recently as an active Emeritus Professor within the RNA Lab of the Department.
Richard was best known for his scientific contributions to understanding the mechanisms and regulation of protein translation initiation. Indeed, he and his PhD student Hugh Pelham developed the widely used reticulocyte lysate system for programmed translation, which enabled in vitro investigations of the mechanisms of translation. Richard's early work as a group leader in the Department was carried out in a very close collaboration with Tim Hunt, with whom he shared a communal lab for many years (see Tim's personal reminiscence below). Together they showed how phosphorylation of initiation factors was a key regulatory step in protein synthesis. Richard's later work focused upon non-conventional 'internal initiation' mechanisms used by various viruses, and on re-initiation from short upstream open reading frames as a regulatory process. Richard was a towering presence in the translation field, widely respected for the clarity and precision of his experimental work and scientific writing, and well-known (possibly even feared) at meetings for his incisive questions based on encyclopaedic knowledge and logic.
Richard's scientific contributions were widely recognised: he was elected as a Member of the European Molecular Biology Organisation (EMBO) in 1991 and as a Fellow of the Royal Society in 2006.
Richard was an incredibly supportive and friendly colleague and mentor and despite all his achievements he was always understated and modest. He mentored a number of PhD students many of whom have gone on to make their own marks as international leaders in their fields. He was also unfailingly supportive of more junior group leaders. After Tim Hunt left the Department, Richard continued the model of a communal lab to draw in younger colleagues with diverse interests in RNA translation, splicing, turnover, editing and localisation, forming the RNA Lab and RNA Biology section of the Department of Biochemistry as part of his legacy.
Richard was incredibly knowledgeable and generous with his time and knowledge. His office was never remarked upon for its organisation, with numerous piles of papers and books and barely a square inch of desk-top visible. However, he had legendary powers of recall; often upon being asked a question he would go directly to the middle of one of these unstable dusty piles and retrieve a paper, which may not have been looked at for many years but that provided the answer to the query.
With Richard's passing we have lost a great friend and scientist.
Richard is survived by his wife Wiltrud, his daughters Tina and Bridget, and grandchildren, who were with him at the end.
Chris Smith and Nancy Standart
Tim Hunt's reminiscences on working with Richard
Richard graduated with his PhD in 1966. It was a two-volume affair that led to an edict that, henceforth, theses should not exceed 50,000 words. In it, he measured the number of tRNAs per active ribosomes and discovered 5S RNA, but published neither as far as I know. So he was finishing his thesis work as I started mine (that was in Michaelmas 1964).
Then Richard disappeared from sight - at one point, Interpol was on the lookout for him - but surfaced in Geneva in Alfred Tissières' laboratory. I don't know what he did there, except that the focus of the lab was on heat shock protein synthesis. I don't think any papers were published, and I don't know how long he stayed there, but the next and first really important thing was his and Tony Hunter's discovery that haemoglobin synthesis was initiated with a special methionine-carrying tRNA. Pretty amazing to have published nothing for 6 years but then get a Demonstratorship (Junior Lecturership) and a full paper in Nature as your first effort as a PI! When he reappeared, it was with his wife Wiltrud.
Richard was extremely pleased to see me back in Cambridge, right at the end of 1970, and we immediately set to work on the question of how haem (and double-stranded RNA) controlled protein synthesis in rabbit reticulocyte lysates. Tony Hunter was around briefly, but went off to resume his tenure at the Salk Institute. So we had a technician, Ann Brayley, and two Graduate students, Chris Darnbrough and Steve Legon, and the lab was sandwiched between Jean Thomas to the East and Richard Perham to the west in the newly-built extension to the old Biochemistry Building, the Wellcome Wing. I had a research fellowship from Clare College that paid about £850 and we made progress when Chris discovered that the peak of methionine label over the 40S subunit disappeared when you added dsRNA to the lysate, and crucially disappeared before the polysomes broke down, so we focused for the first time on this complex, which had been dismissed up to then as a kind of artifact - the 40S ribosomes are 'sticky', people thought. But we convinced ourselves that this was a real preinitiation complex, and invented what we called 'The shift assay' for mRNA, because we could take these things and get them to become 80S ribosomes by adding mRNA. We ran into a lot of scepticism from the reviewers who were utterly fixated on the idea that tRNA only bound to ribosomes as instructed by mRNA. We didn't understand the first thing about protein synthesis! With the help of our friend Mike Mathews, then at the LMB, we showed that the complex did not contain mRNA. But it took years for the idea that the 40S ribosome used its bound Met-tRNAi to locate the start of the message to sink in. I was pleased to see that Nature recently carried an article by Venki Ramakrishnan and his colleagues giving the structure of this and describing the whole process - a gap of almost 50 years!
After that we got rather bogged down. An Israeli group discovered that high levels of 3'-5' cAMP overcame the inhibitions that set in when haem was absent or dsRNA was present and we tested lots of purines - there's a BBRC paper that reveals our bafflement, because these compounds antagonised the inhibitions in subtly different ways. We also looked at the inhibitor edeine and discovered that it blocked the joining of the 60S subunit, a result that was never published because of the fire in our lab in 1974. Fortunately, Steve and Chris had both graduated and left the lab when this happened, and the loss of one rather minor bit of work was more than compensated for by our re-equipment and move up the road to the hospital, within the orbit of the LMB.
It was Richard who with typical tenacity and sublime intelligence discovered that the inhibition of binding of initiator tRNA to 40S ribosomes needed ATP. I never really, properly understood his arguments and experiments, but once we accepted this it was a short step to adding gamma 32P ATP to some purified ribosomes plus and minus dsRNA and seeing a stonking black band on the autoradiogram, right at the molecular weight of one of the subunits of eIF2 (which Theo Staehelin had recently purified). QED! It took a lot of effort to write this all up, and I'm sorry that I do not have a copy of the first edition of this paper before Ben Lewin skilfully edited it down from Richard's detailed and meticulous account.
We moved back downtown, reluctantly, to the Sir William Hardy Building (where the sperm whale who provided John Kendrew with his myoglobin had once 'lived' – it was called The Low Temperature Research Station, LTRS to its familiars, whose other claim to fame, we liked to think, was the invention of the frozen pea). It was here that I suggested that Hugh Pelham (Part II 1975) and Richard try using pre-incubation with micrococcal nuclease, which requires Ca2+, and then chelation with EGTA, to remove endogenous mRNA in reticulocyte lysate, and they totally succeeded in getting the conditions right, and the background of globin synthesis was no longer a concern. This was the basis of the widely used in vitro translation system, and Richard became a consultant for Promega to develop this commercially. We also taught the people at Amersham International how to make lysates and analyse the products on acrylamide gels in return for free [35S]-methionine.
Richard was a really wonderful person to share a lab with, and with whom to collaborate. He perfectly exemplified for me Jim Watson's advice always to work with people who are cleverer than you are. You could show him a result and he immediately grasped its significance, and together we made lots of discoveries that were great fun to work out. Actually, our greatest work is summarised in three papers published in the European Journal of Biochemistry, where we discuss the precise environment of the reticulocyte lysate in terms of its pH, Mg2+, ATP, GTP and spermidine requirements, as well as an admittedly incomplete diversion into thioredoxin and glutaredoxin, using 2'-5' ADP Sepharose filtration. Richard was such a scholar! He would suddenly bring up the existence of the cytoplasmic form of isocitrate dehydrogenase, which unlike its mitochondrial counterpart did not bind to the 2'-5' ADP Sepharose column! How did these crucial nuggets of information lodge in his mind? We never completely understood all that, and neither did we get to the bottom of why protein synthesis needed glucose-6-phosphate apart from its ability to generate NADPH. Business that will remain unfinished, I'm afraid. The wretched editors of the European Journal of Biochemistry refused to accept DTT as an abbreviation for dithiothreitol, and we had to type out "dithiothreitol" 220 times and stick it into the manuscript before it could be accepted. We never published there again.
Something I've always felt, based on these years, roughly 1971-1981 inclusive, is that there is a lot to be said for avoiding responsibility. Richard had a proper job, giving the group a certain legitimacy, but the graduate students and I lived from hand to mouth on short-term grants and fellowships. Poor old Richard was constantly having to lecture or examine or go to a college meeting or something; very difficult to carry out competitive cutting-edge research under such burdens. But it was OK, because I (or Tony Hunter) was in the lab the whole time, so stuff got done. When I started going to Woods Hole in the summer, I was always surprised at how little progress the students made when there wasn't someone to nudge them. The other thing was of course the students - when Hugh Pelham, Richard Treisman and Andrew Murray passed through the lab, Hugh as a graduate student, the other two as 'projectiles' (Part II project students), I for one felt like giving up, because they were so obviously much cleverer than I. But what else could you do? Had to carry on regardless.
Our ways began to diverge in the 1980s, especially after I discovered cyclin and Richard got stuck into polio and IRESs and things. Also, I got a lectureship, but we continued to share the lab and stay in touch. We really wanted to move on from the reticulocyte lysate, but never worked out how; the only outcome of this sally was the discovery of the DNA-dependent protein kinase by Tony Walker and Carl Anderson, another funny story. They were convinced that a particular tRNA was responsible, and it took a lot of badgering to persuade them to test the effects of DNase and RNAse on the preparation of Sigma yeast so-called tRNA they were using to provoke the phosphorylation of what turned out to be a heat shock protein in the HeLa extracts they were using. But our extraordinary ignorance about DNA and all its doings is revealed by our failure to test single-stranded DNA or intact plasmid DNA... so we completely missed the important fact that it was double-stranded ends, stubs of DNA that were responsible!
Richard and my final collaboration was to develop a coupled transcription-translation system using T7 RNA polymerase. I suggested that this might work, I think, to be met with much scoffing and mocking, but then it was tried out as a classroom exercise and once the correct balance of ribonucleotide triphosphates and Mg2+ had been established - a perfect thing for a large class to find - it worked brilliantly.
Richard was a wonderful scientist, a great friend, and a thoroughly good man. And I would say a life well lived.
Tim Hunt
List of key papers, most of them mentioned or alluded to above, in reverse chronological order:
Craig D, Howell MT, Gibbs CL, Hunt T, Jackson RJ (1992). Plasmid cDNA-directed protein synthesis in a coupled eukaryotic in vitro transcription-translation system. Nucleic Acids Res., 20(19):4987-4995. doi: 10.1093/nar/20.19.4987
Jackson RJ, Hunt T (1983). Preparation and use of nuclease-treated rabbit reticulocyte lysates for the translation of eukaryotic messenger RNA. Methods Enzymol., 96:50-74. doi: 10.1016/s0076-6879(83)96008-1
Jackson RJ, Herbert P, Campbell EA, Hunt T (1983). The roles of sugar phosphates and thiol-reducing systems in the control of reticulocyte protein synthesis. Eur. J. Biochem., 131(2):313-324. doi: 10.1111/j.1432-1033.1983.tb07264.x
Hunt T, Herbert P, Campbell EA, Delidakis C, Jackson RJ (1983). The use of affinity chromatography on 2'-5' ADP-sepharose reveals a requirement for NADPH, thioredoxin and thioredoxin reductase for the maintenance of high protein synthesis activity in rabbit reticulocyte lysates. Eur. J. Biochem., 131(2):303-311. doi: 10.1111/j.1432-1033.1983.tb07263.x
Jackson RJ, Cambell EA, Herbert P, Hunt T (1983). The preparation and properties of gel-filtered rabbit reticulocyte lysate protein synthesis systems. Eur. J. Biochem., 131(2):289-301. doi: 10.1111/j.1432-1033.1983.tb07262.x
Farrell PJ, Balkow K, Hunt T, Jackson RJ, Trachsel H (1977). Phosphorylation of initiation factor eIF-2 and the control of reticulocyte protein synthesis. Cell, 11(1):187-200. doi: 10.1016/0092-8674(77)90330-0
Pelham HR, Jackson RJ (1976). An efficient mRNA-dependent translation system from reticulocyte lysates. Eur. J. Biochem., 67(1):247-256. doi: 10.1111/j.1432-1033.1976.tb10656.x
Balkow K, Hunt T, Jackson RJ (1975). Control of protein synthesis in reticulocyte lysates: the effect of nucleoside triphosphates on formation of the translational repressor. Biochem. Biophys. Res. Commun., 67(1):366-375. doi: 10.1016/0006-291X(75)90325-3
Hunter T, Hunt T, Jackson RJ, Robertson HD (1975). The characteristics of inhibition of protein synthesis by double-stranded RNA in reticulocyte lysates. J. Biol. Chem., 250(2):409-417. PMID: 803491
Legon S, Brayley A, Hunt T, Jackson RJ (1974). The effect of cyclic AMP and related compounds on the control of protein synthesis in reticulocyte lysates. Biochem. Biophys. Res. Commun., 56(3):745-752. doi: 10.1016/0006-291x(74)90668-8
Darnbrough CH, Legon S, Hunt T, Jackson RJ (1973). Initiation of protein synthesis: evidence for messenger RNA-independent binding of methionyl transfer RNA to the 40S ribosomal subunit. J. Mol. Biol., 76(3):379-403. doi: 10.1016/0022-2836(73)90511-1
Darnbrough C, Hunt T, Jackson RJ (1972). A complex between Met-tRNAf and its disappearance during incubation with double-stranded RNA. Biochem. Biophys. Res. Commun., 48(6):1556-1564. doi: 10.1016/0006-291x(72)90891-1
Jackson R, Hunter T (1970). Role of methionine in initiation of haemoglobin synthesis. Nature, 227(5259):672-676. doi: 10.1038/227672a0