Wednesday, October 7, 2020

Rosalind Franklin, revealing the the double helix of DNA, the structures in carbon materials and the shapes of viruses

Roaslind Franklin, linocut, 11" x 14", by Ele Willoughby 2020

 

Rosalind Elsie Franklin (25 July 1920 – 16 April 1958), the English chemist and x-ray crystallographer whose x-ray diffraction images were instrumental to discovering the double-helix structure of DNA, has been on my to do list for scientist portraits for years and I've finally got around to it. I think it's the messiness and tragedy of her story that made it a challenge. The version of Franklin which is best known by the general public, is the version presented by the Nobel laureate once described by E. O. Wilson as "the most unpleasant human being I had ever met" James Watson, in his biography The Double Helix, published in 1968. Not only does he call her patronizingly Rosy, he presents her as a dowdy data-hoarding scold who "had to go or be put in her place". Ironically, his biography hurt his own reputation (though he's quite skilled at hurting his reputation more recently, what with the repeated pro-eugenics, racist, homophobic and sexist comments). Watson confessed, “Rosy, of course, did not directly give us her data. For that matter, no one at King's realized they were in our hands.” Strangely, after the publication of their DNA structure papers, Watson and Franklin were on friendly terms, exchanged letters and he even once offered her a lift across the US. His version of her as the villain emerged years after her death, when he wrote the book; some (like Franklin's biographer Maddox) suggest guilty feelings about the irregular way her data was accessed and insufficiently cited are the explanation. Before the book was published, Francis Crick, Maurice Wilkins, Linus Pauling, Max Perutz and Rosalind's brother Colin all protested angrily at her portrayal (as well, often, as the portrayal of themselves and other scientists), especially as she could not defend herself. This forced Watson to add an epilogue praising her as a scientist and claiming he hadn't sufficiently appreciated the experience of women in science at that time. Wilkins wrote to Harvard University Press that the book remained disgraceful and they dropped it; it was published instead by Athenaeum Press, becoming a bestseller.

There's a joke amongst scientists that goes, "What did Watson and Crick discover?" "Rosalind Franklin's notes." And while it's important that her contributions are now recognized posthumously, there's more to her story and to the story of DNA research. Ironically, the famous Photo 51 produced by Franklin's graduate student Raymond Gosling, which Maurice Wilkins quietly shared with Watson and Crick and cemented their thinking about molecular structure, is a photo that Franklin had previously presented at a seminar attended by Watson (where he failed to notice it, presumably thinking patronizing thoughts about "Rosy"). Watson, Crick and Wilkins were awarded Nobel prizes in 1962, after her death.  Though this was prior to the institution of the informal rule against awarding posthumous Nobels, Franklin was not nominated. Before her life was cut tragically short by ovarian cancer, both prior to and after her DNA research, Franklin also made invaluable contributions across disciplines of physics, chemistry and biology, working to determining the structure of RNA, viruses, coal and graphite. Even aside from her role in determining the structure of DNA, her research was a great benefit to society. I wanted to make sure my portrait represented all of this.

But first, this is a wonderful comic by the one and only Kate Beaton:

 

by Kate Beaton
by Kate Beaton from Hark, A Vagrant

Rosalind was born in 1920, the second of five children born to a liberal, affluent and influential London Jewish family. Her father was a merchant banker and his uncle had been Home Secretary, the first practising Jew to serve in the British Cabinet. Her father taught electricity, magnetism and history of the Great War, at the Working Men's College and eventually became Vice Principal. Her family helped settle Jewish refugees fleeing the Nazis, especially children from the Kindertransport. They took two of the children into their home. Rosalind attended St. Paul's school, a leading girls' private school, one of few girls' schools which taught physics and chemistry. She excelled at sciences, languages and sports and won a scholarship for university. Her father asked her to donate the funds to a refugee student. She studied chemistry at Newnham College, Cambridge, completing her undergraduate studies in 1941. Due to the sexist attitudes of the day, women were not granted full degrees, but "degrees titular," until 1948 (when previous women's degrees were retroactively awarded). In her last year at Cambridge she met a French refugee and former student of Marie Skłodowska-Curie, Adrienne Weill; this friendship was an opportunity to practice her French and became important in her career.

She began a PhD project on the polymerisation of acetaldehyde and formic acid under the supervision of Ronald Norrish, Professor of Physical Chemistry at Cambridge and later a Nobel laureate. It was not a fruitful collaboration. His own biographer described Norrish as "obstinate and almost perverse in argument, overbearing and sensitive to criticism" and Franklin grew to despise him and resigned. She gladly took an opportunity to transfer to the British Coal Utilisation Research Association (BCURA) at Kingston-upon-Thames. She began focusing on the porosity and density of coal, to learn how to increase the efficiency of the widely used fossil fuel resource. Her work also had important implications for the effectiveness of the activated-charcoal filters in Second World War gas masks, issued to the entire British populace in case of gas attack. In working to accurately determine the porosity of coal, she made what was likely the first demonstration that coal acts as a molecular sieve (letting helium molecules through but not larger hexane and benzene). This property is still important to industry today.

She completed her doctorate on the structure of carbon materials in 1945 (and once again, as a woman, had to wait to be awarded her full degree until 1948). Her friend Adrienne Weill suggested she attend a Royal Institution meeting in London where she might meet Marcel Mathieu and Jacques Méring. At this conference her interest was sparked in x-ray diffraction and she met the great crystallographer J.D. Bernal (with whom she would later work). Impressed with her paper Méring offered her a researcher post in Paris to continue her work on carbons for four happy years. Méring was a x-ray crystallographer, who employed the way substances diffracted x-rays to deduce their structure and he taught her how to apply this method. By 1950 she had published a paper in Nature about the structure of carbon, and by the following year had learned that as carbon in the form of coal burns, it can form one of two structures: graphitizing and non-graphitizing (terms she coined). In my portrait, the pattern on her jacket is based on her own publication of the structure of non-graphitizing carbon. She showed these two structures explained the difference between the two possible products of burning coal: cokes and chars, and how they burn. This research had important industrial applications. She continued to write papers about the structure of carbon until she died. 

She returned to England in 1950 to work with John Randall, head of the Biophysics Research Unit at King’s College, on a three-year Turner-Newall Fellowship. She planned to look at protein structure but he suggested she work on DNA, as Maurice Wilkins was doing. Randall did not clarify whether Franklin or Wilkins would lead this research, which set their relationship off on the wrong foot. Randall reassigned Wilkin's graduate student Raymond Gosling as her assistant; this surely also did not help things between them. Franklin refined and adjusted the fine-focus X-ray tube and microcamera ordered by Wilkins, improved upon his technique by manipulating the critical hydration of her specimens and employing all her physical chemistry expertise. Wilkins inquired about this and felt her reply was superior. Her directness and enjoyment of a good debate were a bad match for his shyness and distaste for arguments, and their personalities clashed badly.

As early as November 1951, Franklin presented their data at King's College London and noted,

"The results suggest a helical structure (which must be very closely packed) containing 2, 3 or 4 co‐axial nucleic acid chains per helical unit, and having the phosphate groups near the outside."

Franklin and Gosling found there were two forms of DNA: long and thin when wet (dubbed B-DNA) and short and fat when dry (dubbed A-DNA). Because of their conflicting personalities, Randall divided the labour so Franklin and Gosling studied the A form and Wilkins the B. They produced beautiful images of DNA during this time, including Gosling's famous Photo 51 (represented in blue in my portrait). By 1951, the King's researchers all believed B-DNA was a helix, but Franklin felt the evidence for A-DNA was still conflicting. Through painstaking work, by January 1953, Franklin reconciled the conflicting data, concluding both forms had two helices. She drafted three papers, and two noted the double helical DNA backbone. She had also decided to leave the unpleasant atmosphere at King's and move to Birkberk College. Randall insisted that the DNA research stay at King's and Gosling would be reassigned back to Wilkins.

Meanwhile in Cambridge, James Watson and Francis Crick were simultaneously working on the problem and had seen a preprint of Linus Pauling's incorrect proposal for DNA. They came to King's to urge collaboration to win the race before Pauling discovered his error. Thanks to Franklin's identification of the nature of the symmetry of the DNA crystals (that is, its space group), Crick understood that DNA strands were antiparallel (and that both Pauling's model and Watson and Crick's previous model were incorrect). Unable to find Wilkins they spoke with Franklin who was unimpressed by Watson's implication she could not interpret her own data; they argued. Wilkins arrived, commiserated with them, and showed Watson Franklin's work and Gosling's DNA image.  From Wilkins' perspective, Franklin was leaving, and Gosling was now his student; but it seems he did not let Franklin know he had done this. In February 1953, Watson and Crick began working on a molecular model of B-DNA, something Franklin felt was premature, with much of the data based on work at King's. Crick got access to many of Franklin's crystallographic calculations when his advisor Max Perutz gave him a report written for a Medical Research Council biophysics committee visit to King's in December 1952. Though not explicitly marked confidential there was an expectation that such a report would not be shared and Perutz later defended this action with inexperience with administrative matters. Franklin's A-DNA paper was submitted 6 March 1953, one day before Crick and Watson had completed their model on B-DNA. She had not seen Watson and Crick's work when she submitted her paper (though they had of course benefited from seeing her, Gosling and Wilkin's work). Her laboratory notebooks reveal that she had already Franklin noted that ‘an infinite variety of nucleotide sequences would be possible to explain the biological specificity of DNA’ all on her own. Franklin got to see the model built by Watson and Crick on April 10 and apparently commented, "It's very pretty, but how are they going to prove it?" Her philosophy as an experimentalist was to be able to rigourously prove a model correct before publishing. Franklin did modify her paper while in press, after seeing the others' work to acknowledge their work. A trio of DNA papers were published as 25 April 1953 Nature articles. Watson and Crick's paper only acknowledged "having been stimulated by a general knowledge of" Franklin and Wilkins' "unpublished" contribution. Due to agreements between the directors of the King's and Cambridge labs, Wilkins and Franklin published the two other DNA articles with x-ray diffraction data supporting Watson and Crick's model (rather than presenting them as the data which underpinned the model). Watson and Crick invited Wilkins to be a co-author but he declined because he had not help build the model. He later lamented that they can not discussed authorship more thoroughly. 

It may have been less than obvious to Watson and Crick how to cite materials they used, including Photo 51 and the MRC report, but it is something that they could have been done and ethically, many argue that they should have done. Likewise, Watson and Crick were aware of Franklin and Gosling's paper (which included Photo 51), submitted before she saw their work, but they merely noted that their model was not inconsistent.  Any mention they made of Franklin is in combination with, and after naming Wilkins. There is a strong case to be made that Franklin's work was insufficiently credited, and its value and role in determining the structure of DNA has only be recognized posthumously. There's no evidence that Watson and Crick ever let Franklin know what they later acknowledged, that they could not have made their model without out her work, or that she felt insufficiently recognized in their publication. They remained friends during her life and continued to correspond about their respective research projects.

By mid-March, Franklin moved to the much less fancy but much more pleasant Birkberk College, having been recruited by physicist John Desmond Bernal. She was much more comfortable at the non-denominational Birkbeck than the Anglican King's. Though Bernal wanted her to move on from nucleic acid, she continued to mentor Gosling and aid him with the completion of his thesis. The two published the first evidence of the double helix in the A form of DNA in the 25 July issue of Nature. With funding from the Agricultural Research Council, she was able to start her own research group and begin working on RNA and the structure of the Tobacco Mosaic Virus (TMV), an RNA virus. In my portrait her brooch is a model of TMV. She published her first major TMV paper in 1955 in Nature, in opposition of prominent and powerful virologist Norman Pirie, who wrote her angry, condescending letters and refused to ever again supply her with virus samples to study. But her careful observational work was once again correct. With her grad student Kenneth Holmes she found the protein covering was molecules arranged in helices. Early in 1954, she happened to meet Aaron Klug on the stairs at work. She showed him her photo of the TMV and he wrote, it sealed his fate. Captivated, he sought permission to switch to virus research. They began a long and fruitful collaboration. She oversaw a group with her students, Aaron Klug and his student John Finch and the group published on TMV, cucumber virus 4, turnip yellow mosaic virus and other plant viruses. She had a student James Watt supported by the National Coal Board and continuing her longstanding research interest in carbon. With postdoc Donald Caspar she showed that the RNA was wound on the inner surface of the hollow TMV.  She had begun working on the structure of the Polio virus, receiving with Krug the large grant ever at Birkbeck. While traveling in the US in 1956, she noticed her stomach had swelled and she went to the doctor upon her return. They found two tumours. She had ovarian cancer. She continued working when not hospitalized or convalescing with family and friends (including Francis and Odile Crick, with whom she remained close). Her group produced seven papers in 1956 and six in 1957, despite the cancer. She  was promoted to Research Associate in Biophysics on the 25th of February. Tragically she was not able to proceed with the polio research as her health rapidly deteriorated. She was invited to display a large model of TMV in Brussels at Expo 58, the first major international fair after World War II; the fair opened the April 17, but she died April 16, 1958 of bronchopneumonia, secondary carcinomatosis, and ovarian cancer. It's possible that x-ray exposure played a role in the cancer. Science during the 1950s was far too laissez-faire about radiation shielding. The following year, Klug and Finch published the polio virus structure and dedicated the paper to her memory.

By 1962 the scientific community at large was convinced of the structure of DNA, and Watson, Crick and Wilkins were awarded the Nobel Prize in Physiology or Medicine. The rules preclude splitting the award more than 3 ways and Wilkins' inclusion was based not only on his role in discovery but his later work providing data to support the model. As Franklin foresaw, it took years of work to actually convincingly prove the "pretty" model. But, even Watson suggested that Wilkins and Franklin might instead have shared the Nobel in Chemistry. Franklin was never nominated, even though this predates their rule against posthumous prizes.

Also, her long-term collaborator on virus structure, Aaron Klug continued the work he began with Franklin, winning the 1982 Nobel Prize in Chemistry for his development of crystallographic electron microscopy and his structural elucidation of biologically important nucleic acid-protein complexes. Her staunchest defender, and beneficiary of her will, he spoke of her and her impact upon him in his acceptance speech.

Today, she has become one of the most widely recognized researchers in the history of science, with many awards, buildings, plaques and monuments in her honour, worldwide. Elucidating the structure of DNA has perhaps had the most impact on society at large, but her research in carbon and on viruses also has lasting significant impacts in science.  

References

Brenda Maddox,  'Rosalind Franklin, The Dark Lady of the DNA,' HarperCollins, 2002.

Brenda Maddox, The double helix and the 'wronged heroine'. Nature 421, 407–408 (2003). https://doi.org/10.1038/nature01399

Peter J. F. Harris and Irene Suarez-Martinez, 'Rosalind Franklin, Carbon Scientist', Carbon, vol. 171,  January 2021, pp. 289-293 https://doi.org/10.1016/j.carbon.2020.09.022 

'Rosalind Franklin was so much more than the ‘wronged heroine’ of DNA', editorial, Nature 583, 492 (2020)

'Rosalind Franklin,' Wikipedia, accessed October 2020.

Mathew Cobb, 'Sexism in science: did Watson and Crick really steal Rosalind Franklin’s data?' The Guardian, Tuesday 23 June, 2015. 

, Rosalind Franklin

 Dainton, Sir Frederick Sydney (1981). "Ronald George Wreyford Norrish, 9 November 1897 – 7 June 1978". Biographical Memoirs of Fellows of the Royal Society. 27: 379–424. doi:10.1098/rsbm.1981.0016. JSTOR 769878. S2CID 72584163

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