Fred Sanger (1918–2013): a Science Hero

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Obituary

Fred Sanger (1918–2013): a science hero

But the outcome of Sanger’s life in research military service and applied to do research was much greater, and, as the late Guy Dodson with N.W. (‘Bill’) Pirie, the proteins expert, so aptly put it, “…it was an unequalled on obtaining edible protein from grass. By the foundation for the most prodigious explosion time the bucketful of ground-up grass Pirie that is modern biology”. His unerring kept in the deep-freeze had thawed, Pirie had chemical judgement, skills as an experimenter left Cambridge for Rothamsted. and tenacity are legendary, as was his modesty. Fortunately, Albert Neuberger, who had His many honours included, in addition to the just arrived in Cambridge as a postdoc, took

Nobels, an FRS (1954), Foreign Associate of Sanger on as his PhD student. In 1943, he Downloaded from http://portlandpress.com/biochemist/article-pdf/36/2/42/3096/bio036020042.pdf by guest on 02 October 2021 the National Academy of Science, USA (1967), completed his thesis on ‘Lysine metabolism CBE (1963), CH (1981) and OM (1986). and the more practical problem concerning the He had apparently declined a knighthood nitrogen of potatoes’. As he once put it to me, because, he said, he didn’t like the idea of being “he [Neuberger] was the one who really taught called ‘Sir’… me how to do research, both by example and His most public legacy is no doubt the instruction”, claiming that he only did what Sanger Centre, later the Wellcome Trust Neuberger told him to do “as it took quite a Sanger Institute, at Hinxton Hall, Cambridge, time – about two years in my case – before you where a significant part of the human genome start to think out your own experiments”. He project was carried out. John Sulston, its first also felt that the valuable experience he gained Director, vividly recalls that when he asked on amino acid chemistry during his doctoral Fred Sanger in about 1958 (Peter Sanger) him over the phone whether it could be called work on lysine was a good introduction to his the Sanger Centre, Sanger replied “Well, okay, future work on ‘sequences’. but it better be good!” Fred Sanger, who died on 19 November 2013, Frederick (Fred) Sanger was born in the From lysine metabolism to the first was one of the most remarkable scientists of Gloucester village of Rendcombe in 1918 complete sequence of a protein our time. He worked at the bench between where his father, a practising medical doctor 1943 and 1983, the first 20 years or so at the who had done research in Cambridge, greatly After completing his PhD, Sanger joined A.C. Biochemistry Department, University of stimulated his interest in science as well Chibnall’s research group before becoming a Cambridge, on amino acids and proteins, as inculcating in him, through a Quaker member of the External Scientific Staff of the resulting in the complete sequencing of insulin upbringing, the principles of pacifism and Medical Research Council (1951). Chibnall had in 1955, the first ever protein to be sequenced. the value of truth. Although he abandoned just succeeded F.G. Hopkins as Professor and It was this achievement that earned him the religious aspects of Quakerism by the Head of Biochemistry at Cambridge in 1943. his first (and solo) Nobel Prize for Chemistry time he was 19, he remained a conscientious His group’s main expertise was quantitative in 1958. objector and committed supporter of anti- analysis of the amino acid composition of Over the next two decades he went on, at nuclear campaigns throughout his life. A proteins, especially insulin, one of the few the MRC Laboratory for Molecular Biology, to major influence on his choice of career was proteins readily available at the time in pure devise methods for sequencing nucleic acids, his chemistry teacher at Bryanston, who form. According to Sanger, it was Maurice culminating in 1977 in the development of an encouraged Sanger to ‘play around’ in the Rees, Chibnall’s chief assistant, who discovered ingenious method for sequencing DNA (the chemistry laboratory where he enjoyed that insulin had more free amino acids than ‘Sanger dideoxy method’), which opened the experimenting with dyes and, especially, the could be accounted for by its lysine content, way to the complete sequencing of the human growing of beautifully coloured crystals. leading Chibnall to suggest that Sanger look genome in 2001. For this he was awarded his Sanger first came across ‘biochemistry’ into this: the rest is history! Sanger first went on second Nobel Prize in Chemistry in 1980 as an undergraduate at Cambridge, where he to invent fluorodinitrobenzene (later known as which he shared with Walter Gilbert, “for chose it as a final fourth-year option, inspired the Sanger reagent) as a new coloured reagent contributions concerning the determination by talking with Ernest Baldwin, his supervisor to label the N-terminal groups of insulin of base sequences in nucleic acids”, the other at St. John’s College. He was apparently quite and, in 1945, after two years of painstaking half going to Paul Berg for his work on surprised at obtaining a first class degree work he was able to identify of glycine and recombinant DNA. Fred Sanger was one of in 1940 (qualifying him for postgraduate phenylalanine as the N-terminal amino acid only four Nobel laureates ever to be awarded research) as he ‘had not done too well in residues of what were later identified as the two two prizes, and alone in receiving them both exams’. Being a Quaker and registered as a cross-linked chains of insulin. There followed for chemistry. conscientious objector, he was exempted from the crucial and serendipitous discovery

42 April 2014 © Biochemical Society Obituary

(reported in 1949 in the Biochemical Journal1) The second phase: sequencing had developed that allowed ‘reading off’ that mild (as opposed to complete) hydrolysis nucleic acids sequences directly from their position on of 2,4-dinitrophenol (DNP)-labelled insulin autoradiographs. Further refinements of produced short sequences of four or five Sanger carried out much of his research on these ingenious techniques led to Sanger and amino acids near the N-termini that could proteins in an overcrowded basement laboratory his colleagues to sequence much longer RNA, be separated by paper electrophoresis, within the Biochemistry Department. Through e.g. the 3300-nucleotide-long bacteriophage producing a distinct pattern which Sanger his enthusiastic encouragement, he was R17RNA. As some of the sequences matched called a “fingerprint”. (Sanger frequently instrumental in getting Max Perutz to persuade the codons predicted from the amino acid

acknowledged his debt to Archer Martin and the Medical Research Council (MRC) to fund sequence of the coat protein (already known), Downloaded from http://portlandpress.com/biochemist/article-pdf/36/2/42/3096/bio036020042.pdf by guest on 02 October 2021 Richard Synge for introducing the methods of a new building to accommodate both their it enabled them for the first time to directly partition chromatography that he was able to research groups. And in about 1962, he began determine (and thus confirm) the genetic exploit so successfully for this work.) Sanger the second phase of his career in the newly code from a nucleotide sequence. considered his 1949 ‘peptide paper’ as the most created MRC Laboratory of Molecular Biology. significant of his early papers on insulin as it Here he led the Protein and Nucleic Acid The ultimate challenge: DNA demonstrated for the first time that sequences Chemistry Division (initially called the Protein could, in principle, be located in a protein Chemistry Division) until 1983, when his good Then��–��and apparently to the dismay of some molecule. That was quite a revolutionary friend and colleague Cesar Milstein took over. people, including Dorothy Hodgkin who felt concept at the time, when most biochemists Although Sanger had previously felt he should have returned to proteins��–��Sanger believed the arrangement of different amino little interest in nucleic acids, after moving committed to elucidating the sequence of acids in a protein to be random. Importantly, to the LMB, he conceded that “with people DNA, that of the bacteriophage ΦX174 to partial hydrolysis became a central general like Francis Crick around, it was difficult to start with. This must surely be counted as approach to the sequencing of proteins, and ignore nucleic acids or to fail to recognize the one of the boldest leaps forward ever taken later of nucleic acids. importance of sequencing them”3. An even in biology. An American reviewer consulted By 1955, through work with two of his more important influence was apparently about Sanger’s ambitious new programme postdocs, Hans Tuppy and later Ted Thompson, John Smith, a very co-operative colleague who said, “Fred Sanger continues to bang his head Sanger had succeeded in revealing, as one taught him how to fractionate nucleotides. against brick walls. And the brick walls keep writer put it, “the linear world of polypeptides”, George Brownlee, who joined him as his falling down.” at the same time as Benzer was revealing the first PhD student in 1963, somewhat surprised Sanger initially developed copying linear world of genes. The fact that there Sanger by choosing to work on RNA, rather methods with DNA polymerase and short was a well-defined sequence in proteins led than on proteins – “a bold choice at the nucleotide primers to successfully derive a directly to the idea that there had to be a time!”†. Sanger considered this a strong factor 50-nucleotide sequence of the single-stranded genetic code*. In addition, in one of several in getting his whole group to move in that f1 bacteriophage DNA. He and Alan Coulsen papers published in the Biochemical Journal general direction. then went on to develop a fast read-out in 1955 completing the work on insulin, Initial investigations on nucleic acids method, the so-called ‘plus and minus method’ Sanger and his colleagues discovered discrete were with defined RNA structures, e.g. and, shortly thereafter, the game-changing species-specific variations in amino acid tRNA and rRNA, and explored the use of sequencing methodology, the ‘dideoxy chain sequence within the ‘glycyl chain’ of insulin. using RNA-cleaving enzymes to prepare termination method’, used to confirm their This provided the first evidence for significant partial digests. This led to the discovery that earlier sequence for the ΦX174 DNA (some similarities between proteins that could radiolabelled di-, tri- and tetra-nucleotides 5375 nucleotides long). This was the first be assumed to be related evolutionarily could be separated by 2D ionophoretic whole genome sequence ever produced and (i.e. homologous), “an insight that led directly techniques. The first real breakthrough, Sanger’s group went on to sequence the 16 569 to the development of sequence alignment according to Sanger, came when Brownlee bases of the human double-stranded mtDNA methods, still the most commonly used succeeded in deciphering the 120-nucleotide (1981): the first extensive human DNA to bioinformatics tools”. For this, he has been sequence of 5S rRNA from Escherichia coli, be sequenced. Sanger then developed the credited as the “grandfather” of bioinformatics2. using novel 2D separation techniques they so-called ‘whole-genome shotgun method’ he

* One of the first persons who made that connection early on was the young Cambridge chemist, John Griffith, who Sanger once told me he considered “a bit of a genius”. † For text references to Sanger and references to key papers, see Guy Dodson’s ‘BJ Classic Papers’ article (doi:10.1042/BJ2005c013) and my 2005 interview at www.biochemistry.org/AboutUs/Archive/Archivecollection/FredSangersequencingpioneer.aspx

sequencing work….[as] a form of relaxation after receiving his letter in 2005 inviting compared with the more original, exacting, me to collect all (35) of his, mostly hand- variable, and often frustrating, work on written laboratory notebooks which he had development of methods”4. Whatever he felt decided many years earlier to donate to the were his own strengths, Sanger never failed to Biochemical Society for its archives‡. I wish fully acknowledge the contributions of all of someone had photographed both of us scaling the people he interacted with, at whatever level. the ladder to the loft and gingerly descending Sanger, once referred to as the most with arms laden with the notebooks! focused of men, was passionate about research I later had the privilege of interviewing and about the intense enjoyment he got from him at his home, wondering once again how “messing about in the lab”. As he once put it, “of this amazingly self-effacing, quiet and civilized

the three main activities involved in scientific person was indeed the same, some would Downloaded from http://portlandpress.com/biochemist/article-pdf/36/2/42/3096/bio036020042.pdf by guest on 02 October 2021 research, thinking, talking and doing, I much say visionary, Fred Sanger whose impact on prefer the last and am probably best at it”3. the course of biology has been likened to John Sulston once remarked in an interview Charles Darwin’s. Contemporary science that Sanger “…was a quiet guy, he just got historians are quite rightly reluctant to speak on with it – that was his style …I found him of heroes, but I hope they might make an completely inspirational”. exception with Sanger. He was also most unusual in deciding to Fred Sanger died at Addenbrooke’s Hospital give up research completely after retiring from on 19 November 2013. He married Joan Howe the LMB in 1983, at the statutory age of 65, as (who died in 2012) in 1940, and is survived by he felt his life’s work had been completed with his three children and two grandchildren. ■ the sequencing of DNA. The ending of his life in the laboratory was certainly abrupt and I thank Professor George Brownlee for allowing decisive, as related by John Finch in his history me to ‘mine’ the obituary he published in Current Fred Sanger’s letter to the Biochemical Society of the LMB: “The conventional finishing date Biology 23, R1074–R1076 archivist. in the MRC is 30 September, and on that day Fred was doing experiments – on 1 Oct his John Lagnado and Coulson used to sequence the genome of laboratory was empty and he was gone”4. (Honorary Archivist) the virus bacteriophage λ (48 502 nucleotides) With characteristic modesty, Sanger later in 1982. commented that he would have felt guilty at occupying space that could have been References Fred Sanger, a modest genius available to a younger person and that he 1. Sanger, F. (1949) Biochem. J. 45, 563–574 looked forward in retirement to spending 2. Samson, C. (2006) The Biochemist, 28 (6), 48–49 Although often referred to as the father of time enjoying his family, ‘messing about 3. Sanger, F. (1988) Annu. Rev. Biochem. 57, 1–29 genomics, Sanger always considered his own with boats’ and tending the lovely garden at 4. Finch, J. (2008), A Nobel Fellow on Every Floor: contribution to nucleic acid sequencing was his home in the Cambridgeshire village of a History of the Medical Research Council primarily to the methods. He wrote that he Swaffham Bulbeck. Laboratory of Molecular Biology, Medical “quite enjoyed the somewhat routine DNA I had the opportunity of visiting Fred Research Council, Cambridge

‡ The Biochemical Society archives, including the notebooks and several audio- and video-tape interviews of Sanger, were catalogued in 2009 and are kept on permanent loan at the Wellcome Library. It is particularly pleasing that the Wellcome has included Sanger’s notebooks (and related correspondence) as part of their ambitious digitizing project, making their content readily accessible online.