PROFILE Profile of Joachim Messing PROFILE Brian Doctrow, Science Writer Joachim Messing’s father did not initially approve of cholesterol and fatty acid me- his son’s decision to become a scientist. “He was a tabolism. Lynen advised Mess- mason,” Messing explains, “and was hoping that I ing not to go into Lynen’sown would just take over his business.” His mother, how- field, where much of the im- ever, was more supportive, “because she didn’t like portant work had already been theroughclimateontheconstructionsites.” Fortu- done. Lynen then told Mess- nately for science, his mother prevailed. Messing, ing that the future would be the director of the Waksman Institute of Microbiology in DNA, and arranged for Mess- and the Waksman chair of Molecular Genetics at ing to work in a laboratory Rutgers University, was elected to the National Acad- studying DNA at the Max Planck emy of Sciences in 2015. The DNA sequencing tech- Institute for Biochemistry in nique he developed made possible the subsequent Munich. Joachim Messing. Image courtesy of Michael sequencing of whole genomes, including the human Seul (photographer). genome. Beginning of Shotgun Sequencing From Pharmacy to Laboratory For his graduate research, Messing studied DNA repli- Joachim Messing, known as “Jo” (pronounced yoh), cation, and got off to what he calls “a lucky start.” At that grew up in the town of Duisburg, Germany, in the time, it was known that the enzyme DNA polymerase aftermath of World War II. The first person in his family required a primer to begin DNA replication, but the to go to college, he decided to study pharmacy as an identity of the primer was unknown. Early on in his undergraduate. At that time, a degree in biochemistry graduate research, Messing learned about a plasmid was not an option for undergraduates. Pharmacy that could replicate in the absence of protein synthesis. appealed to Messing, in part, because the curriculum He quickly demonstrated that the same plasmid covered a broad range of subjects, including organic could not replicate in the absence of RNA synthesis, and inorganic chemistry, botany, and even physics. suggesting that RNA acted as a primer for DNA po- “You had to learn how to look under the microscope lymerase. “That was probably the first evidence of a and draw cells,” Messing recalls, “you had to learn noncoding RNA in vivo,” Messing explains. The find- about the different pathways in plants that make ing was published in Nature New Biology, only three important substances that could be used for ther- months after Messing began his graduate research (1). apy...you had everything.” Studying pharmacy would Messing received his doctorate in biochemistry from also allow him to earn money to pay for his education the Ludwig Maximillian University of Munich in 1975. by working as a pharmacist. After that time, he stayed on as a research fellow at the For the first two years of the pharmacy curriculum, Max Planck Institute, where he began the work that Messing had to work as a pharmacist’s apprentice. would lead to shotgun sequencing. In 1974, while still The pharmacy where he did his apprenticeship was working on his doctorate, Messing attended a confer- across from the German Opera on the Rhine in Duis- ence on restriction enzymes in Ghent, Belgium, where burg. In the evenings, after working at the pharmacy, Frederick Sanger reported on his efforts to sequence he would work as an extra with the opera company. the single-stranded genome of bacteriophage ΦΧ174. The high point of his opera career, Messing says, was Sanger was using a time-consuming procedure that getting to carry the actress playing Gilda down a involved cleaving the double-stranded version of the flight of stairs in a production of Rigoletto. phage DNA with a restriction enzyme, separating the Messing received a bachelor’s degree in pharmacy fragments by gel electrophoresis, extracting each frag- in 1968 and a master’s degree in pharmacy in 1971. ment from the gel, denaturing the fragments, and using When he decided to pursue a doctorate in bio- the denatured fragments as primers on the viral single- chemistry, he met with the biochemist Feodor Lynen, stranded DNA for sequencing by DNA synthesis. Messing who had won the Nobel Prize in 1964 for work on thought that the sequencing process could be accelerated This is a Profile of a recently elected member of the National Academy of Sciences to accompany the member’s Inaugural Article on Page 7949. www.pnas.org/cgi/doi/10.1073/pnas.1608857113 PNAS | July 19, 2016 | vol. 113 | no. 29 | 7935–7937 Downloaded by guest on October 4, 2021 by cloning each fragment into a single-stranded DNA improved levels of the essential amino acids lysine and phage vector. In this way, all of the fragments could be methionine, and to pest-resistant crop strains. sequenced in parallel, each using the same primer. In 1983, the Waksman Institute of Microbiology at There was just one obstacle—a suitable vector did Rutgers University sent Messing an offer to become not yet exist, so Messing had to create one. A suitable their new research director. Recognizing the name, vector had to provide a means of rapidly detecting Messing dug through some old photographs and cells containing the cloned inserts. For this purpose, found one of himself as an undergraduate with Selman Messing developed the blue-white screen, in which a Waksman, the institute’s namesake, who had won the gene on the vector turns bacterial colonies blue when Nobel Prize in 1952 for methods leading to the dis- grown on a special medium unless the vector contains covery of streptomycin, the first antibiotic treatment a cloned insert. The first intermediate to incorporate for tuberculosis. “So I said to my wife, ‘oh, you know, the visible marker, M13mp1, was published in PNAS maybe I should have a look,’” he recalls. Messing de- in 1977 (2). Next, Messing had to insert an artificial cided to come to Rutgers because he felt it was a place restriction site for cloning DNA fragments into the where he could really make an impact. Messing arrived visible marker gene. The gene contained a sequence at Rutgers in 1985, where, in addition to taking over that differed from a restriction site by only one nu- the directorship of the Waksman Institute, he was also cleotide; it had a G instead of an A. Methylation of tenured as a university professor. the G base induced mispairing, which converted the G to A in subsequent rounds of replication. “I think the New Frontiers biggest moment in all the research that I’ve done was Today, Messing continues to expand the possibilities of certainly the day when I went into the darkroom to genome sequencing. In his Inaugural Article (6), he re- look at...whether the restriction site was there or not ports testing a single-molecule sequencing platform in the right place,” Messing recalls. “It was unusual from the sequencing company Pacific Biosciences that timing because, a week later, I had to go to the Cold can sequence very long stretches of DNA in the maize Spring Harbor phage meeting... and you never have genome. Eukaryotic genomes contain many duplicate things that work in time [for meetings]... but in this sequences, which prevent the reassembly of entire case, it really worked like a charm.” The results were chromosomal regions from sequenced fragments if the published in Nature in early 1978 (3). fragments are too short. The new platform overcomes this difficulty and could become important for studies of Move to the United States and Plant Genetics the human genome and of diseases such as cancer. In 1978, Messing moved to the University of California, According to Messing, the greatest challenge was San Francisco for a few months, and then to the University finding a computer cluster powerful enough to handle of California, Davis, where he continued to refine his the massive amounts of data produced. Ultimately, shotgun sequencing system. With the help of researchers Messing and his colleagues had to enlist the help of at the biotechnology firm Genentech, he developed Amazon’s computing services. The study illustrated that synthetic oligonucleotide primers and used synthetic advances in sequencing have come not only from the DNA adapters to introduce multiple restriction sites into development and implementation of new techniques the phage vector, allowing different types of restriction but also from progress in computing power. fragments to be cloned (4). In the first successful dem- Messing’s contributions to DNA sequencing and onstration of the shotgun sequencing method, Messing plant genetic engineering have won him numerous and a plant pathologist, Robert Shepherd, sequenced the awards, including the 2013 Wolf Prize in Agriculture 8-kb genome of the cauliflower mosaic virus (5). By that and the 2014 Promega Biotechnology Research time, Messing had taken a faculty position at the Uni- Award. In 1991, the Institute for Scientific Information versity of Minnesota. in Philadelphia declared him the world’s most cited Meanwhile, Messing began applying the techniques scientist of the previous decade. Messing gets great he had developed to the study of plant genetics. “Isaw satisfaction from the knowledge that his work has left and right how people said ‘well, the future is going made possible subsequent life-saving developments, to be in eukaryotic DNA, you can’t keep working with such as the production of recombinant erythropoietin microbes,’” Messing recalls. Studying plants appealed for treating anemia following chemotherapy. “There is to him, in part, because of his pharmacy background, no doubt,” he says, “that genome sequencing and and also because of the potential practical applications engineered genes had an unforeseen impact on pro- for nutrition.