Vol.Vol. 19 19 / / No. No. 5 4 / / May April 2020 2020

THETHE MEMBERMEMBER MAGAZINEMAGAZINE OFOF THETHE AMERICANAMERICAN SOCIETYSOCIETY FORFOR BIOCHEMISTRYBIOCHEMISTRY ANDAND MOLECULARMOLECULAR BIOLOGYBIOLOGY

The COVID-19 issue A new career center Browse jobs, post positions, have your resume critiqued and more.

careers.asbmb.org NEWS FEATURES PERSPECTIVES 2 22 50 EDITOR’S NOTE A LEGACY OF TYROSINE ON THE FRONT LINE: Breaking the news PANDEMIC INSIGHT FROM 3 A HEALTH CARE WORKER MEMBER UPDATE COVID19 52 30 A small army of researchers 7 races to build a QUARANTINED THOUGHTS IN MEMORIAM interactome 32 Could an old malaria drug 54 10 help fight SARS-COV02? JOURNAL NEWS A NEW CITY, A NEW JOB 10 as a detective’s assistabt 34 Anatomy of a molecule: what AND A GLOBAL PANDEMIC 12 Cow born in Japan after removal, makes remdesivir promising? replacement of placental cells 13 How is myelin made? 36 Slipping past the proofreader 16 Review delves into 59 44 Scientist uses community proximity proteomics IS MORE SCIENCE THE MEDICINE organizing skills to mobilize 17 From the journals WE NEED TO CURE THE WORLD’S researchers against COVID-19 STRUGGLING ECONOMY? 46 Researchers retool genomics labs to provide 2 testing 48 “We are doers. We want to get involved.” 12 22 13

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MAY 2020 ASBMB TODAY 1 EDITOR’S NOTE

THE MEMBER MAGAZINE OF THE AMERICAN SOCIETY FOR BIOCHEMISTRY AND MOLECULAR BIOLOGY Breaking the news OFFICERS COUNCIL MEMBERS By Comfort Dorn Gerald Hart Suzanne Barbour President Joan Broderick Toni M. Antalis Matt Gentry President-elect Blake Hill Audrey Lamb Wei Yang James M. Ntambi Secretary Takita Felder Sumter Joan Conaway Kelly Ten–Hagen Treasurer JoAnn Trejo

EX-OFFICIO MEMBERS ASBMB TODAY EDITORIAL ADVISORY BOARD Robert S. Haltiwanger ON UNSPLASH MIKA BAUMEISTER BY PHOTO Rajini Rao Carla Koehler Chair Co-chairs, 2020 Annual Ana Maria Barral Protective masks, normally used for surgery, are now in use to fight the coronavirus, SARS-CoV-19. Meeting Program Committee Natasha Brooks Cheryl Bailey Kelly Chacón Chair, Education and Beronda Montgomery his month marks my third an- aware of an insidious new disease that Professional Development Bill Sullivan niversary as managing editor of was sickening people in China. We Committee Melissa Vaught ASBMB Today. If COVID-19 posted our first article about research Daniel Raben Binks Wattenberg T Chair, Meetings Committee restrictions continue, I’ll probably related to chloroquine and “the new Sonia Flores ASBMB TODAY celebrate privately with an extra cup coronavirus” on Feb. 6, covering a Chair, Minority Affairs Angela Hopp Committee Executive Editor of coffee at my kitchen table/desk. paper published two days earlier. Nicole Woitowich [email protected] Maybe even a doughnut. Since then, this job and my old Chair, Science Outreach and Comfort Dorn As I’ve mentioned in this space jobs have felt increasingly similar as Communication Committee Managing Editor Terri Goss Kinzy [email protected] before, I worked for about 20 years we race to share news of COVID-19 Chair, Public Affairs Lisa Schnabel at daily and weekly newspapers research and how our members are Advisory Committee Graphic Designer before I came to the ASBMB to edit helping and coping. Our staff writ- Ed Eisenstein [email protected] Chair, Membership Committee John Arnst a monthly magazine. The change was ers, contributors and members have Susan Baserga Science Writer stark, mostly in terms of pacing. I churned out articles and essays at [email protected] Chair, Women in Biochemistry spent much of my career hounding what, for us, is an astonishing pace. and Molecular Biology Laurel Oldach Committee Science Writter reporters to turn around daily stories Why am I telling you this? [email protected] Sandra Weller in a matter of hours. Here, I found This issue of ASBMB Today Chair, Publications Ed Marklin Committee Web Editor that we had the luxury of working for reflects these recent changes. Here [email protected] Lila M. Gierasch weeks, even months, on articles and we have collected the best of the Editor-in-chief, JBC Allison Frick Multimedia and Social Media essays to get them exactly right for COVID-19 writing that we’ve posted A. L. Burlingame Content Manager this magazine. Very little was urgent. on our website since February. We Editor, MCP [email protected] We moved at a stately and thoughtful tried to update articles wherever Nicholas O. Davidson Barbara Gordon Editor-in-chief, JLR Executive Director pace, rather like an ocean liner or a possible, but the story is evolving [email protected] Kerry-Anne Rye dowager countess. quickly. This is a snapshot, from the Editor-in-chief, JLR That changed this year. And it viewpoint of this magazine and this changed fast. society, of a moment that comes once For information on advertising, contact Pharmaceutical Media Inc. at 212-904-0374 or [email protected]. First, we moved to daily pub- in a century. lishing on our website in January, Stay well and stay safe. meaning we were posting fresh stories every morning — often several a day.

This had been a longtime goal of the Comfort Dorn www.asbmb.org/asbmbtoday ASBMB Today staff, and we were ([email protected]) is the PRINT ISSN 2372-0409 managing editor of ASBMB delighted with our fresh new website, Today. Follow her on Twitter @cdorn56. Articles published in ASBMB Today reflect solely the authors’ views and not even though it meant major pivots in the official positions of the American Society for Biochemistry and Molecular our workflow. Biology or the institutions with which the authors are affiliated. Mentions of products or services are not endorsements. Later that month, we became

2 ASBMB TODAY MAY 2020 MEMBER UPDATE

Three ASBMB members win Protein Society awards

Pictured, from left, are Protein Society award winners Catherine Drennan, Stephen Sligar and Karen Fleming.

The Protein Society has honored three members of lipid-membrane patches stabilized by a belt of mem- the American Society for Biochemistry and Molecular brane-scaffolding proteins. For that work, he also won Biology with 2020 awards. Karen Fleming of Johns the ASBMB’s 2016 Herbert A. Sober Lectureship, Hopkins University, Stephen Sligar of the University of recognizing outstanding biochemical and molecular Illinois at Urbana–Champaign and Catherine Dren- biology research, with a special emphasis on the devel- nan of the Massachusetts Institute of Technology were opment of methods and research techniques. set to receive their awards at the World Conference on The Anfinsen award is sponsored by the Protein Protein Science in June, which was canceled in response Society. ASBMB members who have won it in the past to the COVID-19 pandemic. include Anthony Kossiakoff, University of Chicago Karen Fleming, a professor of biophysics at Hop- (2019); Sachdev Sidhu, University of Toronto (2015); kins and a pioneer in the study of membrane-protein and Barry Honig, Columbia University (2012). folding, won the Carl Brändén Award, which honors Catherine Drennan, a professor at MIT and a a protein scientist who has contributed significantly to Howard Hughes Medical Institute investigator, won science, education and/or service. the Dorothy Crowfoot Hodgkin Award for solving Fleming is an associate editor for the Journal of high-resolution structures of proteins and protein Biological Chemistry, served on the ASBMB Council complexes that enhance understanding of the biology of from 2014 through 2017 and co-founded the Gordon metalloproteins. Research Conference on Membrane Protein Folding. Hodgkin used X-ray crystallography to determine She recently received the Society of General Physiolo- the structure of vitamin B12 and won the 1964 Nobel gists’ inaugural Sharona Gordon Award. Prize in chemistry. The Brändén award is sponsored by Rigaku Corp. The Protein Society, in its announcement, pointed ASBMB members who have won it in past years to Drennan’s determination of the first structure of co- include Billy Hudson, Vanderbilt (2017); C. Robert balamin-dependent ribonucleotide reductase, one of the Matthews, University of Massachusetts (2015); Stephen three enzymes that catalyze the final step in production White, University of California, Irvine (2014); Sheena of deoxyribonucleotides in all organisms. Radford, University of Leeds (2013); Helen Berman, Drennan is a former member of the ASBMB Rutgers University (2012); Michael Summers, Univer- Education and Professional Development Committee, sity of Maryland, Baltimore County (2011); and Bruce a past ASBMB annual meeting session organizer, and a Alberts, University of California, San Francisco (2010). past winner of the ASBMB–Schering–Plough Research Stephen Sligar, who chairs the biochemistry depart- Institute Award. ment at UIUC, won the Christian Anfinsen Award for The Hodgkin award is sponsored by Genentech. methodological advances in the field of protein sciences. ASBMB members who have won it include Hao Wu, Anfinsen received the in chemistry in (2019); Susan Marqusee, 1972 for his work on enzyme structure. His namesake University of California, Berkeley (2018); Manajit Hay- award recognizes “a technological achievement or sig- er–Hartl, Max Planck Institute of Biochemistry (2017); nificant methodological advances in the field of protein Rachel Klevit, University of Washington (2016); Judith sciences.” Frydman, (2014). Sligar discovered and developed nanodiscs,

MAY 2020 ASBMB TODAY 3 MEMBER UPDATE

University of Vermont names interrogate the protein machines and a Presidential Early Career for Parise as dean involved in repairing DNA and Scientists and Engineers for his Leslie Parise, maintaining integrity. metabolism research. In one line of a biochemist who is The International Award — one inquiry, his lab uses novel systematic currently chair of of 10 awards given each year by the screening to glean functional insights the department of United Kingdom–based Biochem- into orphan mitochondrial proteins. biochemistry and ical Society — recognizes research Other lines of inquiry concern the biophysics at the that illustrates the importance of the synthesis of coenzyme Q and the role University of North molecular biosciences in the advance- of phosphatases in regulating mito- Parise Carolina, Chapel ment of life sciences and is given to chondrial activities. Hill, and until recently was chair of early-career scientists of any nation- Since 2015, in addition to the faculty at that university, will ality who are located outside the UK running his lab, Pagliarini has also di- soon join the faculty of the Universi- and Ireland. rected the metabolism branch of the ty of Vermont. There, she will serve “This certainly would not have independent Morgridge Institute for as the dean of the College of Agricul- been possible without the hard work Research, a nonprofit institute affiliat- ture and Life Sciences. and creativity of the members of ed with UW-Madison and located on Parise’s lab focuses on the molec- my laboratory,” Eichman said of the its campus. ular mechanisms of and blood award in a statement. “I thank my BJC Investigators are funded clot formation. One line of inqui- colleagues who made and endorsed by the St. Louis-based hospital and ry focuses on the protein calcium the nomination and who have guided physician organization BJC (original- and integrin-binding protein 1, or me over the years, and my family ly Barnes–Jewish/Christian) Health- CIB1, which may be a drug target for their support and encourage- Care. They are selected by a search in triple-negative breast cancer cells; ment. This award will help maintain committee of 42 professors at Wash- a second research program has to do momentum within my research team, ington University School of Medicine. with the role of a lipid deacetylase in so that we may continue to approach Pagliarini is the fifth of 10 professors platelet aggregation. what we feel are important and in- the program is projected to recruit. In addition to her service at teresting questions related to genome UNC Chapel Hill, Parise was an maintenance.” Olzmann wins associate editor for the journal Blood. presidential award She has served as president of the Pagliarini to move University Association of Medical and Graduate from Madison to Missouri of California, Departments of Biochemistry and on Dave Pagliarini, Berkeley faculty the American Society for Biochem- an associate profes- member James istry and Molecular Biology’s public sor of biochemistry Olzmann was affairs advisory committee. She is at the University of among the 315 re- scheduled to start at UVM in May. Wisconsin–Madison cipients of the best known for his Olzmann 2019 Presidential Eichman wins award work on defining Early Career Award for Scientists and Pagliarini from Biochemical Society and functionalizing Engineers. Vanderbilt Uni- mitochondrial proteins, has been Olzmann, an associate professor versity biochemistry named a BJC investigator and will in the department of nutritional stud- professor Brandt join the faculty at Washington Uni- ies and toxicology and an investigator Eichman has won versity in St Louis in May. at the Chan Zuckerberg Biohub, the Biochemical Pagliarini received the Earl and investigates the regulation of lipid Society’s 2021 In- Thressa Stadman award from the droplets and the role that they play ternational Award. American Society for Biochemistry in maintaining lipid homeostasis in Eichman Eichman’s lab and Molecular Biology this year and conditions such as obesity and fatty uses structural biology techniques has previously landed young investi- liver disease. He joined the faculty at including X-ray crystallography to gator awards from the Protein Society UC Berkeley in 2013 after earning

4 ASBMB TODAY MAY 2020 MEMBER UPDATE

his Ph.D. from Emory University and Fuchs and Bissell win 2020 Gairdner awards doing postdoctoral work at Stanford University. Established in 1996, the PECASE honors the contributions scientists and engineers have made to the advancement of science, technol- ogy, education, and mathematics education and to community service as demonstrated through scientific leadership, public education and community outreach. Outstanding investigator award for UW’s Bornfeldt Mina Bissell and Elaine Fuchs Karin Born- feldt, a professor Elaine Fuchs and Mina Bissell are among the five scientists to receive of medicine and a 2020 Canada Gairdner International Award. pathology at the The award honors Fuchs, a pioneering cell biologist, for revealing the University of mechanism by which cells make and repair tissues. Much of what Washington and we know about human skin’s capacity to heal and regenerate — and, in an associate editor cases of mutation, to succumb to diseases like epidermolysis bullosa — has Bornfeldt of the Journal of been made possible by Fuchs’ work, from her first lab at the University of Lipid Research and several other Chicago to her current position as the Rebecca C. Lancefield investigator journals, has received an Outstanding at the . Investigator Award from the National The award honors Bissell, a distinguished senior scientist at the Heart, Lung and Blood Institute. Department of Energy’s Lawrence Berkeley National Laboratory, for her Diabetes can accelerate the pro- paradigm-shifting work in modeling the two-way interactions between cess of atherosclerosis, wherein cho- normal and malignant cells in tumor microenvironments. This behavior, lesterol-rich lesions form in arterial called “dynamic reciprocity,” revealed that tumor cells behave differently walls. This puts people with diabetes in lab environments than in patients, led to 3D understanding of tumor at heightened risk of heart attack and behavior and has had significant impacts on cancer therapies. stroke. Bornfeldt’s lab studies the role The Gairdner Foundation was established by Canadian businessman of lipids, lipoproteins and immune and philanthropist James A. Gairdner in 1957 with the goal of recognizing cells in this process. In addition to and rewarding international excellence in fundamental research that im- running a research lab, she directs pacts human health. The foundation has bestowed 395 awards on laureates the Diabetes Complications Program from 35 countries, 92 of whom have gone on to receive Nobel prizes. This and serves as associate director for re- year’s International award winners also include Roel Nusse, Rolf Kemler search of the UW Medicine Diabetes and Masatoshi Takeichi. Institute and as deputy director of the UW’s Diabetes Research Center. The Outstanding Investigator Award mechanism provides substantial flexible funding for a research program, Marion B. Sewer Distinguished rather than supporting specific projects. Scholarship for Undergraduates Bornfeldt and her team will receive $7.2 million over up to seven years to Deadline: June 1 pursue strategies for preventing cardio- asbmb.org/diversity/undergraduate-scholarship vascular complications of diabetes.

MAY 2020 ASBMB TODAY 5 MEMBER UPDATE

Student chapter members land Goldwater scholarships

The recipients of this year’s Goldwater scholarships • Mark Hargrove, a professor at Iowa State Uni- were announced in late March. versity, working with Behnia Rezazadeh Shirazi The scholarships, named in honor of Senator Barry • Matthew Francis and , pro- Goldwater, offer $7,500 per remaining academic year fessors at the University of California, Berkeley, to sophomore and junior students pursuing bachelor’s working with Casey Mogilevsky degrees in the natural sciences, math or engineering. • Leslie Hicks, an associate professor at the Many of this year’s recipients in the life sciences University of North Carolina at Chapel Hill, are American Society for Biochemistry and Molecular working with Lauren Lim Biology Student Chapter members. The ASBMB con- • Terry Hill, a professor at Rhodes College, work- gratulates these promising sophomores and juniors: ing with Ramiz Somjee • Furyal Ahmed, Agnes Scott College • Vincent Hilser, a professor at Johns Hopkins • Chris Bragança, Villanova University University, working with Andrew Munoz • Daniel Cheong, University of Oklahoma • Michael Jewett, a professor at Northwestern • Landon Clark, University of Georgia University, working with Alexandra Wooldredge • Madeline Farringer, Iowa State University • Henrik Kibak, a professor at California State • Shellaina Gordon, Northeastern University University, Monterey Bay, working with Saman- • Edena Khoshaba, Chapman University tha Miller • Stella Ma, University of Wisconsin–Madison • Daniel Kraut, an associate professor at Villano- • Eran Maina, The College of Wooster va University, working with Chris Bragança • Emily Mahoney, Rochester Institute of Technol- • C. Martin Lawrence, a professor at Montana ogy State University, working with Sheridan Brown • Rishi Mehta, University of Cincinnati • Teresita Padilla-Benavides, a professor at the • Mlana Lore, Eckerd College University of Massachusetts Medical School, • Jessica Pierce, Salisbury University working with Shellaina Gordon • Cynthia Schofield, University of Massachusetts • Tanya Paull, a professor at the University of Boston Texas at Austin, working with Cassandra Bishop • Ramiz Somjee, Rhodes College • Subbiah Pugazhenthi, an associate professor at • Daniel Wieland, University of Arizona the University of Colorado Anschutz Medical • Karen Zhang, University of Washington Campus, working with Anit Tyagi • Jacob Schwartz, an assistant professor at the As part of the application, students write an essay University of Arizona, working with Daniel about their experience with their research mentors. Wieland ASBMB salutes the many mentors of this year’s 287 • Vincent Tagliabracci, an assistant professor at Goldwater scholars in the life sciences, who helped the University of Texas Southwestern Medical ignite their students’ excitement about biology. Among Center, working with Patrick Nnoromele the ASBMB members who mentored a Goldwater • Timothy Wencewicz, an associate professor at scholarship recipient are: Washington University in St. Louis, working • Jeanine Amacher, an assistant professor at Western with Michael Moore Washington University, working with Min Gaoå • James West, an assistant professor at The Col- • Pascale Charest, an associate professor at the lege of Wooster, working with Eran Maina University of Arizona, working with Jamison • Crystal Young-Erdos, an assistant professor at Takashima Eckerd College, working with Mlana Lore

6 ASBMB TODAY MAY 2020 IN MEMORIAM

Jerry Lingrel Bruce S. McEwen

Jerry Lingrel, a Neuroscientist longtime associate Bruce S. McEwen, who editor of the Journal of made important discov- Biological Chemistry and eries about the effects of a University of Cincinnati stress and sex hormones College of Medicine fac- on the brain, died Jan. 2. ulty member for almost He was 81. 60 years, died Feb. 22. McEwen, the Alfred He was 84. E. Mirsky professor In a 2007 and head of the Harold autobiographical and Margaret Millikin paper, Lingrel wrote that as a child he “spent a lot of time Hatch Laboratory of Neuroendocrinology at The Rockefeller watching things grow and being fascinated by the complexity University, worked at the intersection of neurobiology, endo- of the structure of different flowers, trying to understand how crinology and behavioral science. His research changed the engines worked … I had a real curiosity about science, but had way scientists conceived of the brain. no idea what real experimental science involved.” Encouraged “When he began his career in the 1960s, most scientists by an enthusiastic biology teacher, he went to Otterbein Col- believed that the brain ceases to change when it becomes fully lege where he majored in biology and chemistry, then earned a developed, at which point its basic architecture becomes sta- Ph.D. in biochemistry from The Ohio State University followed ble,” explained Rockefeller writer Katherine Fenz. “Research of by a postdoc at the California Institute of Technology. the day focused largely on ion movements and the transmis- Lingrel joined the faculty of the University of Cincinnati in sion of chemical signals across synapses. But McEwen, along 1962 and became a professor of biochemistry and molecular with a few other scientists, recognized that the brain is in fact biology in 1972. He served for 27 years as chair of the depart- malleable and can be modified by circulating hormones.” ment of molecular genetics, biochemistry and . McEwen discovered adrenal steroid-binding sites in After stepping down in 2008, he returned to serve as the the brain, showed that brain regions not previously linked to department’s interim chair from 2014 to 2017. hormone or stress regulation, including the hippocampus and Lingrel was known for taking a biochemical approach to prefrontal cortex, selectively bind radiolabeled corticosterone, physiological problems. Recently, his research had focused and studied glucocorticoid influences on neuroplasticity. on a zinc finger transcription factor called Kruppel-like factor McEwen was born in Fort Collins, Colorado, on Jan. 17, 2, or KLF2, that is required for vascular integrity and might 1938. He grew up in Ann Arbor, Michigan, where his father was play a protective role against atherosclerosis. In years past, a professor at the University of Michigan. his lab cloned the Na,K-ATPase, a major transport protein, and He earned his bachelor’s degree in chemistry at Oberlin identified specific amino acids involved in binding sodium and College in Ohio and then his Ph.D. in cell biology from Rockefeller potassium as well as those necessary for binding the cardiac University in 1964. He did postdoctoral work in Sweden and took a stimulant molecule ouabain. faculty position for a short time at the University of Minnesota. Lingrel’s career spanned disciplines, he wrote, and He returned to Rockefeller as a faculty member in 1966. “while biochemistry has been a very important aspect of this He was promoted to professor in 1981 and served as associate work, I think my research has always had as a focus the use dean and then dean of the graduate training program between of chemical principles to understand physiological function.” 1985 and 1993. He was named the Mirsky professor in 1999. He also stressed the impact of working with others, writing, McEwen won many awards and honors during his career. He was “we probably have more influence on the individuals we train elected to the National Academy of Sciences, the National Academy of than we anticipate and clearly mentoring should be taken very Medicine and the American Society of Arts and Sciences. seriously.” He is survived by his wife, neuroimmunologist Karen Bulloch, with whom he collaborated; his former wife, Nancy, and their two daughters, Carolyn and Sarah; stepchildren, Kimberly McGrath and Scott Muryasz; and eight grandchildren.

7 ASBMB TODAY MAY 2020 IN MEMORIAM

Michael Wakelam James C. Hu

Michael Wakelam, a James C. Hu, a professor at Cambridge longtime professor University and an in the Texas A&M associate editor for the University department ASBMB’s Journal of Lip- of biochemistry and id Research, died March biophysics, died in 31 of respiratory failure, his home in College likely from COVID-19 Station, Texas, on Jan. . He was 64. 23 from complications Wakelam studied of liver disease. He lipids’ structural, meta- was 66. bolic and signaling roles and pioneered the use of high-sensi- Born April 3, 1953 in Berkeley, California, Hu grew up tivity liquid chromatography–mass spectrometry to measure in Palo Alto. He earned a B.S. in biology at Stanford Univer- lipid levels in various cell and tissue types. sity and a Ph.D. in molecular biology from the University He had been director of the Babraham Institute, a life of Wisconsin. He worked as a postdoctoral fellow at the science research center near Cambridge, since 2007 and a Massachusetts Institute of Technology before establishing his JLR associate editor since 2013. lab at Texas A&M. In a joint statement, JLR’s editors-in-chief, Kerry-Anne In his research, Hu worked to develop biological Rye and Nicholas O. Davidson, said this about Wakelam’s ontologies, sets of standardized controlled vocabularies for personal qualities and work for the journal: “Those who had annotation. Most recently, his lab worked with other groups on the privilege of knowing him enjoyed his generous spirit as well the Ontology for Microbial Phenotypes, a resource for reuse as his passion for science. Michael maintained the highest and analysis of microbial genetics data. His group worked standards of objectivity in his role as an (associate editor) and with the Gene Ontology Consortium on annotation of gene was an exceptional judge of scientific merit.” functions and developed systems for integrating annotation Wakelam was born in 1955. He earned his undergraduate with education in the Community Assessment of Community and doctoral degrees at Birmingham University in 1977 and Annotation with Ontologies. They also worked on systems for 1980, respectively. He went on to complete postdoctoral building model organism databases for community annotation, training at Glasgow University and worked in Germany and including EcoliWiki, which uses and modifies the open-source London before joining Glasgow as a lecturer in 1985. In software built for Wikipedia to provide specialized scientific 1993, he returned to Birmingham as a professor of molecular data resources. pharmacology. Along with his work in microbial genomics and commu- The director position lured him to Babraham in 2007. nity annotation, Hu was known as a supportive colleague who There, he oversaw a team of researchers engaged in preclinical was generous with his time and energy. He loved teaching and and clinical studies of the cellular and molecular details of interacting with students. He mentored undergraduate and aging. graduate students in the lab, at meetings and in collaborative He is survived by his wife, Jane, and their two grown projects. children. Hu is survived by his wife, Deborah Siegele, a professor Editor’s note: In May 2019, ASBMB Today science writer in the biology department at Texas A&M. Generations of Laurel Oldach wrote a cover story about efforts to harmonize students and colleagues have known them as the rare couple lipidomics. For that story, she interviewed Wakelam and others who collaborated at work and at home, in symbiosis yet with involved with the Lipid Metabolites and Pathways Strategy, fierce independence. known more commonly as LIPID MAPS. Read the story.

8 ASBMB TODAY MAY 2020 IN MEMORIAM

Manford K. “Bud” Patterson Jr. Paul Starr Sypherd

Manford K. Paul Starr Sypherd, “Bud” Patterson Jr., a microbiologist who a former officer of held leadership roles the Samuel Roberts at the University of Noble Foundation and California, Irvine and the former president of the University of Arizona, Oklahoma Academy of died Jan. 18. He was 83. Science, died Jan. 22 in Born Nov. 16, Edmond, Oklahoma. He 1936, in Akron, Ohio, was 93. Sypherd moved to Arizo- Born Aug. 20, na at age 6. He earned 1926 in Muskogee, Oklahoma, Patterson served in the a B.S. in microbiology at Arizona State (College) University, an U.S. Navy Air Corps after high school, then earned a B.S. in M.S. in microbiology at the University of Arizona and a Ph.D. at chemistry and an M.S. in biochemistry from the University of Yale University. He was a postdoctoral fellow at the University Oklahoma. He worked for several years in the soils lab of the of California, San Diego and later said he believed his training Noble Foundation, an agricultural research institution, before and experiences at UCSD set him on his academic trajectory. earning a Ph.D. in biochemistry from Vanderbilt University. He Sypherd began his career at the University of Illinois; he served as a consulting biochemists for the Interdepartmental had a fully funded research laboratory and obtained tenure Committee on Nutrition for National Defense before returning there before he was recruited to the University of California, to the Noble Foundation in 1973 as vice president and direc- Irvine where he served as chairman of the department of tor of the biomedical division. Following his retirement from microbiology and molecular genetics for more than a decade, the foundation, he joined IMTEC Corporation as a senior vice director of the medical scientist program, vice chancellor for president of research and development. research and dean of graduate studies. He returned to the Patterson played a role in developing L-asparaginase, a University of Arizona in 1993 as executive vice president and drug used to treat acute lymphocytic in children. provost, leading development of an integrated learning center. He developed a quality control system that enabled IMTEC to As provost, he focused on improving staff and faculty working market in the U.S. He co-edited a book, “Tissue Culture: Meth- conditions. One of his lasting contributions was closing the ods and Applications,” contributed chapters to other scientific campus between Christmas and New Year’s, so employees books, published more than 60 articles in scientific journals could spend time with their families. He retired in 2002 and and held three patents. He was an adjunct professor at was designated provost emeritus and professor emeritus of Oklahoma University and served the American Tissue Culture molecular and cellular biology at the UA. Association as an officer and editor-in-chief of its journals. He Over the course of his 35-year research career, Sypherd played an active role in many scientific and civic organizations and his students published more than 150 articles in peer-re- and served as a judge at numerous international science and viewed journals, many on protein synthesis and nucleic acid engineering fairs. metabolism in and fungi. He was also a pioneer in Patterson is survived by his wife of 66 years, Beverly; a ribosome research. He was a founding editor of the Journal daughter, Shelley Goetz, and her husband, Dan; and a grand- of Molecular and Cellular Biology and editor for the Journal son, Manford Goetz, and his wife, Julie. of Bacteriology, and he served on the National Institutes of Health study section for microbial chemistry for two terms and on the National Research Council committee on medical education. Sypherd is survived by his wife, Linda; his daughter, Denise; sons David, Sean and Scott; his grandchildren; and many other relatives.

MAY 2020 ASBMB TODAY 9 JOURNAL NEWS

Yeast as a detective’s assistant Susan Henry’s work on inositol-containing phospholipids

By Martin J. Spiering

acromolecules such as proteins, “Yeast is almost like the Esche- lipids and carbohydrates often richia coli of the eukaryotic world,” Mhave complex structures that Henry said, adding that yeast helped ASBMB TODAY underpin their cellular functions. researchers “to figure out exactly The sugar alcohol myo-inositol is where the metabolic components are a notable exception — its simple coming from.” six-carbon structure looks unremark- The fully sequenced S. cerevisiae able but is used in countless cellular genome did not become available processes in all domains of life. until 1996, so earlier studies of the According to Susan Henry, a genetics and biochemistry of even professor of molecular biology and this simple organism required skilled genetics at Cornell University, inosi- detective work to find all the players tol is absolutely essential in cells. “It’s involved in a molecular pathway. a major phospholipid precursor as At Albert Einstein College of Susan Henry’s work on the regulation of inositol well as a signaling molecule,” Medicine in the mid-1970s, Henry and phospholipids in yeast enabled numerous she said. and Ph.D. student Michael Culbert- studies of the same pathways in humans and Henry has studied inositol me- son used the mutagenic agent ethyl other eukaryotes. tabolism in the yeast Saccharomyces methanesulfonate to generate more cerevisiae since the 1970s. “I focused than 50 S. cerevisiae mutants de- on the phospholipids and the metab- fective in inositol biosynthesis. This the late 1980s and early 1990s and olites that regulate their formation,” mutant strain collection provided a now recognized as Classics, Henry she said. “Inositol turned out to be resource to launch investigations into and colleagues at Albert Einstein the strongest regulatory metabolite the genes involved in inositol and College and at Carnegie Mellon of these pathways.” phospholipid metabolism. University reported the sequence Early in her career, Henry chose In a 1981 JBC paper, Henry and genetic analysis of the INO1 to work with S. cerevisiae, she said, and co-author Thomas Donahue gene along with its regulation by a because it is easier to study inositol reported the first purification and transcriptional repressor and two and phospholipids in this organism characterization of yeast myo-inosi- transcriptional activators. than in more complex eukaryotes. tol-1-phosphate synthase, or Ino1, In the first of these papers, an enzyme that catalyzes a reaction Henry and Margaret Dean–Johnson Making a mutant resource that yields inositol 1-phosphate, an sequenced the cloned INO1 gene Yeast species have been a work- immediate precursor to free inositol. and also determined the amino acid horse for scientists since the dawn of The two scientists also mapped sequence of the purified protein. This modern research. Their widespread its gene to a locus called INO1 in analysis uncovered an open reading use in fermentations led to the coin- the yeast genome and developed frame, or ORF, as a prime candidate age of the term “enzyme” (Greek for antibodies for specific detection for encoding the entire enzyme. “in yeast”). S. cerevisiae grows rap- of Ino1, laying the groundwork When they disrupted the pre- idly in culture as single cells, a boon for more detailed biochemical and dicted INO1 ORF in yeast cells, the for investigating eukaryotic biochem- genetic studies. researchers found that the cells did istry. The species can be maintained not express the Ino1 protein and stably in the haploid state, and its The first classic grew only when supplied with inosi- genes can be manipulated easily. In three JBC papers published in tol from the growth medium.

10 ASBMB TODAY MAY 2020 JOURNAL NEWS

Their findings showed that the OPI gene in the yeast genome, uncovering how phospholipid syn- INO1 gene encodes myo-inosi- cloned and sequenced it, and identi- thesis is regulated in eukaryotes. tol-1-phosphate synthase in yeast and fied several features of the predicted made available the full-length nucleo- Opi protein sequence, including a A ladder for research tide and amino acid sequences of this leucine repeat and polyglutamine Looking back, Henry says that pivotal phospholipid enzyme. stretches present in many other regu- mentorship by Seymour Fogel and In keeping with earlier findings latory proteins. Alec Keith laid the foundation for her of the Henry lab that expression of The paper defined an important career. Besides sharing their expertise the Ino1 enzyme is transcriptionally regulatory mechanism controlling in genetics and biochemistry, they regulated, their work revealed several INO1 expression. It also provided gave Henry critical material support conserved short DNA motifs in the critical momentum for identifying an to get her work off the ground. 5’ region of the INO1 gene important regulatory DNA element, “I was really lucky that they were that likely were bound by transcrip- the inositol-sensitive upstream not the kind of people who wanted tional regulators. activation sequence. This element is me to (work exclusively) on their hot Henry next set her sights on present in the promoters of genes for project,” she said. “They were willing deciphering the regulation of INO1 biosynthesis of phospholipids and the to let me come into the laboratory expression by inositol and another lipid triacylglycerol. and use their materials to do the phospholipid precursor, choline. The third Classics paper further things that I wanted to do.” Using various INO1 promoter con- elucidated the regulatory circuit that Moreover, although she was structs fused to the E. coli lacZ gene controls phospholipid biosynthe- working with yeast, she secured fund- to measure the promoters’ activities, sis in yeast. Henry and colleagues ing through agencies that typically her team pinpointed the main regula- demonstrated that the yeast proteins support mainly medical research. tory regions in the INO1 promoter. Ino2 and Ino4 form a heterodimeric “I didn’t have any trouble getting In particular, the team found a complex that binds and activates support from the National Institutes region that appeared to be bound by the INO1 promoter and delineated of Health, because of the connec- a transcriptional repressor, Opi1, they the binding sites of the Ino2–Ino4 tion with lipid metabolism in other had previously identified. complex on this promoter. eukaryotic organisms,” she said. The paper was the first to describe This investment paid off. “Many The regulatory circuit a basic helix-loop-helix, or bHLH, of the genes that I worked on were In the second Classics paper, heterodimeric transcription factor in homologous to those in other eu- Henry and colleagues mapped the yeast and represented a milestone in karyotes, providing a ladder for other people to find the (corresponding) genes in other organisms,” Henry said. ASBMB TODAY

Martin J. Spiering ([email protected]) is the technical editor at the Journal of Biological The sugar alcohol myo-inositol is a signaling compound and a ubiquitous precursor to phospholipids Chemistry. Follow him on Twitter @spieringmj. and many other important structural and signaling metabolites.

MAY 2020 ASBMB TODAY 11 JOURNAL NEWS

Cow born in Japan after removal, replacement of placental cells By John Arnst

esearchers at Hokkaido Universi- ty have found that cow embryos Rfrom which placenta-forming cells had been removed can regrow those cells, form a placenta and successfully gestate. The scientists recently published their results, which provide insight into the regenerative capacity of mammalian embryos, in

the Journal of Biological Chemistry. HOKKAIDO UNIVERSITY COURTESY OF NANAMI KOHRI/ All mammalian embryos follow the same blueprint in the first week of development: After being fertilized, a zygote divides into two cells, which Scientists in Japan removed placenta-forming cells from a bovine embryo, but it was able to regrow quickly become four, eight, then 16 those cells, form a placenta and successfully gestate. It is now this healthy 23-month-old cow. The cells that specialize into an inner cell researchers named the cow Matoryona because as an embryo it resembled a Russian matryoshka mass and outer cells that are known nesting doll. individually as trophoblasts and col- lectively as the trophectoderm. Laboratory of Animal Breeding and the weaker regenerative capacity. Nanami Kohri, the lead author Reproduction previously had isolated Kohri and colleagues plan to on the paper, was intrigued by the bovine inner cell masses from embry- investigate what drives the differenc- fact that mouse embryos in which the os at the early blastocyst stage to find es in embryonic protein expression trophoblasts — which differentiate where the genes that give rise to the among mammals as they continue to to form the placenta — had been trophectoderm were being expressed. monitor their calf, which is now 23 removed were much less successful in Other groups had shown that cells months old and healthy. regenerating a placenta than bovine positioned at the outer margin of “It has been suggested that the embryos that also had trophoblasts the inner cell mass could be trans- molecular basis of determining removed. formed into trophectoderm in mouse cellular divisions and localization in “Although isolated inner cell embryos. development differs among species,” masses in both mice and cattle To understand why the bovine Kohri said. “In the future, we will underwent trophectoderm regenera- embryos had more success regener- have to use our experimental system tion, they were significantly different ating placental cells than the murine to evaluate trophectoderm regener- in terms of regeneration efficiency, embryos, the researchers at Hokkaido ation from the reformed inner cell marker protein distribution and University investigated the expression masses in mice and cattle.” expression status of key genes,” he of the gene SOX17, which creates a DOI: 10.1074/jbc.RA119.010746i said. “Surprisingly, a calf was success- protein that regulates cell specializa- fully delivered after the transfer of tion in development. They found that John Arnst (jarnst@asbmb. the reformed inner cell mass to the the expression of SOX17 varied sig- org) is an ASBMB Today science writer. Follow him surrogate mother, but no descendants nificantly between the two species and on Twitter @arnstjohn. were obtained from reformed inner was localized to the trophectoderm cell masses in mice.” cells that had been originally absent in Kohri and his colleagues at the murine embryos, which might explain

12 ASBMB TODAY MAY 2020 JOURNAL NEWS

How is myelin made? Understanding the protective coating on neurons may inform future therapies

By Nuala Del Piccolo

yelin is the protective lipid sheath wrapped around a JENNIFER SIOW Mnerve. It functions as an insula- tor, akin to the protective coating on a wire, speeding up electrical trans- mission of signals along a neuron. Myelin also plays a role in maintain- ing the health of neurons. Myelin function is dysregulated in many neurological disorders, including multiple sclerosis. Oligodendrocytes are the my- elin-producing cells of the central nervous system. The myelin sheath around a neuron is part of an oligo- dendrocyte’s plasma membrane, and a single oligodendrocyte can myelin- ate as many as 50 neurons. During myelination, an oligodendrocyte stretches out tubes of membrane in search of a neuron. When it finds one, it sends the necessary building materials down the tubes and, still operating from a distance, assembles a myelin sheet around the neuron. Composition, number of wraps and total coverage all matter. A myelin- ated neuron that loses its coating cannot transmit electrical signals properly, leading to loss of mus- cle control and other neurological problems. The myelin sheath is mostly made of lipids, including sphingo- lipids, which are critical to myelin’s structure and function. The enzyme serine palymitoyltransferase, or SPT, produces the backbone of all sphin- golipids, and the membrane-bound protein ORMDL monitors sphingo- This artist’s rendition shows neurons with (bottom) and without (top) a myelin lipid levels and regulates SPT activity. sheath. The neuron with a myelin sheath functions at full capacity, while a neuron ORMDL’s activity must be precise: without myelin is unhealthy. Oligodendrocytes are depicted in red.

MAY 2020 ASBMB TODAY 13 JOURNAL NEWS

Too little sphingolipid production during myelination, Wattenberg and measurable, but the team deduced impedes myelination, and too much Sato–Bigbee’s team worked with that ORMDL isoform expression can be toxic. newborn brains, because peak varies over time. These findings pave Binks Wattenberg, a professor of myelination occurs directly after the way for future experiments. biochemistry and molecular biology birth. Only one in five cells in the “The control of sphingolipid at Virginia Commonwealth Univer- brain is an oligodendrocyte, so the biosynthesis is key to myelination, sity, studies membrane biogenesis team isolated these myelin-produc- and understanding how this process and now focuses on lipid biogenesis. ing cells for their experiments. works will enable us to alter it in “I am very curious about how the The researchers found that a future treatments,” Wattenberg said. cell knows when to make sphingo- large portion of the sphingolipids “Our pie-in-the-sky goal is to under- lipid and when to stop,” Wattenberg present in oligodendrocytes during stand sphingolipid biosynthesis so said. “I think ORMDL might be the myelination have an atypically long well that we can reprogram oligoden- key to answering that question.” backbone — an 18-carbon chain drocytes and reverse demyelination Wattenberg’s next-door lab instead of a 16-carbon chain. “The in degenerative myelination diseases neighbor, Carmen Sato–Bigbee, a 18-carbon chain backbone points to like MS.” professor in the same department, a change in lipid composition during DOI: 10.1194/jlr.RA120000627 studies myelination, with a focus on myelination, which might explain oligodendrocytes. The two joined the insulating properties of myelin,” forces to study the role of sphingo- Wattenberg said. “In future work, we Nuala Del Piccolo is a lipid biosynthesis in myelination in want to look at the role of each type postdoctoral scholar in the biomedical engineering developing brains. They report their of sphingolipid in myelination.” department at the recent results in the Journal of Lipid The study also found that SPT University of California, Research. activity increases for the first few Davis. She earned her Ph.D. in materials science To uncover the dynamics of days of myelination and then begins and engineering at Johns sphingolipid content and synthesis to decrease. ORMDL activity is not Hopkins University.

Upcoming ASBMB events and deadlines

MAY

12–18 National Women’s Health Week 20 World Autoimmune Inflammatory Arthritis Day 27 Virtual event: Enzyme regulation by filamentation and other alternate and emerging mechanisms 28 Virtual event: Shape shifting in the control of protein function

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1 Deadline for Marion B. Sewer Distinguished Scholarship for Undergraduates

14 ASBMB TODAY MAY 2020 MAY 2020 ASBMB TODAY 15 JOURNAL NEWS

Review delves into proximity proteomics

By Laurel Oldach

n a recent review article in nearby proteins with a chemical tag The second enzyme family, the ­Molecular & Cellular Proteomics, that then can be purified. After puri- peroxidases, evolved to convert IPayman Samavarchi–Tehrani and fication, mass spectrometry identifies hydrogen peroxide to water by redox colleagues in the Gingras lab at Sinai the tagged proteins. chemistry. In the presence of a bio- Health Systems and the University Most often, the chemical tag is bio- tin–phenol substrate and hydrogen of Toronto offer an introduction to tin, a cofactor that is key to carboxylase peroxide, they can make a short-lived proximity-dependent biotinylation, a enzyme activity in several metabolic free radical that reacts with certain key first step in proximity proteomics. pathways. Two types of enzyme are amino acid side chains, once again The authors give researchers who are used for proximity-dependent bioti- tagging nearby proteins for later new to the field information about nylation: peroxidases, used for methods identification. the natural history of biotinylation such as APEX, and biotin ligases, used As proximity proteomics has enzymes. They also offer insights into for methods such as BioID. grown in popularity, both types of the mechanisms of these enzymes and Ordinarily, biotin ligases append enzyme have been the targets of new perspectives on future proximity biotin to the carboxylases that need extensive engineering and molecu- SAMAVARCHI–TEHRANI ET AL./MCP SAMAVARCHI–TEHRANI

A schematic diagram shows the proximity proteomics workflow. A bait protein is tagged with a biotinylation enzyme (center of concentric circles), which allows for covalent labeling of proteins in its vicinity with a reactive biotin intermediate. Then the researcher lyses the cells and uses streptavidin to extract biotin-tagged proteins, digests those proteins and uses mass spectrometry to determine their identity.

proteomics experiments. it as a cofactor. Biotin ligases found lar evolution to coax them toward Traditional proteomics can pro- in cells have high specificity for the activity profiles users want. The vide information about the quanti- their substrate proteins, but certain authors review the available enzymes tative contents of a cell or tissue, but mutations reduce that specificity by and discuss experimental design con- it sacrifices much information on the decreasing the ligase enzyme’s affinity siderations, such as choice of control spatial organization of proteins with- for a reactive intermediate. Such conditions and how to get rid of what in cells. Since protein activity often mutants lose their grip on the cofac- they call “frequent flyer” proteins that depends on location and interactions tor and can release a reactive biotin often are isolated nonspecifically. with other proteins, researchers have that can bind the next amine group DOI: 10.1074/mcp.R120.001941 developed approaches such as prox- it encounters — often on a nearby imity proteomics to obtain informa- protein. When researchers pull down Laurel Oldach (loldach@ tion about the environs of a protein biotin after this reaction occurs, they asbmb.org) is a science writer for the ASBMB. Follow of interest. Proximity proteomics can determine what proteins were her on Twitter @LaurelOld. methods developed in the past 10 localized in the neighborhood of the years depend on fusing the protein of biotin ligase and, by extension, the interest to an enzyme that will label protein it was tethered to.

16 ASBMB TODAY MAY 2020 JOURNAL NEWS

From the journals

By Latavia Hill, Kian Kamgar-Parsi & Anand Rao

We summarize a selection of various iron pathways to acquire iron HDL have failed. According to Ka- recent papers from the Journal of and effectively compete with other trin Niisuke and a joint team from Biological Chemistry, the Journal bacterial species. Tufts University and the Centers for of Lipid Research and Molecular & DOI: 10.1074/mcp.RA119.001829 Disease Control and Prevention, our Cellular Proteomics. limited knowledge of the composi- A new therapeutic target tion–function relationship in HDL How a pathogen for inflammatory CRMO particles could be a reason for those adapts to survive The autoinflammatory bone failures. In new research published Pseudomonas aeruginosa, a disease chronic recurrent multifocal in the Journal of Lipid Research, Gram-negative opportunistic patho- osteomyelitis, or CRMO, is mediated the researchers sought to elucidate gen, causes burn wound by the inflammatory cytokine inter- better the role of lipid composition and pneumonia. It produces sidero- leukin 1 beta, or IL-1beta, a known in HDL function. phores to acquire iron for survival driver of bone lesions in CRMO. Using samples from CHD and can express various iron-uptake To develop effective therapies for patients and healthy controls, pathways with specific TonB-depen- the treatment of CRMO, research- Niisuke and colleagues were able to dent transporters, or TBDTs, which ers need to better understand the separate out large and small HDL allows it to use exosiderophores pro- signaling events that result in high particles and characterize their duced by other bacterial species. levels of IL-1beta. Tejasvi Dasari lipid compositions. Compositional Quentin Perraud of the Uni- and colleagues at St. Jude Children’s differences were found between the versity of Strasbourg and a team of Research Hospital used mutant large and small particles in all patient researchers in France recently pub- mouse strains, immunoblotting groups, but differences in both lipid lished work in the journal Molecular and microcomputed tomography composition and the function of & Cellular Proteomics focused on to reveal a role for the nonreceptor apoA-1 between healthy subjects and understanding how P. aeruginosa se- spleen tyrosine kinase, or SYK, in the CHD patients were primarily in the lects, regulates and adapts its expres- signaling cascade, resulting in elevat- large particles. These large particles sion levels of iron-uptake pathways ed IL-1beta levels. influence cholesterol control through in response to environmental stimuli. Their work, published in the the scavenger receptor class B type In this study, the researchers showed Journal of Biological Chemistry, 1 pathway; thus, this research not that P. aeruginosa uses different suggests that SYK may be a thera- only provides a deeper understand- siderophores at different chelating peutic target for the treatment of ing of the composition–function efficiencies, with catechol sidero- CRMO. relationship of HDL but also could phores being the most powerful. DOI: 10.1074/jbc.RA119.010623 inform development of treatments They also showed that expression of for CHD. the TBDTs varied when P. aeruginosa Good cholesterol gone bad DOI: 10.1194/jlr.RA119000258 was grown in three different media, in coronary heart disease which suggests that different pheno- High-density lipoprotein, com- A new drug’s role in typic patterns exist. This work also monly called HDL, often is referred cholesterol trafficking shows that P. aeruginosa can detect to as “good cholesterol” for the Niemann–Pick type C disease, the presence of epithelial cells and beneficial role it plays in maintaining or NPC, is a fatal genetic disorder adjust gene expression accordingly. cholesterol balance in the body. HDL that results in the accumulation of These findings show that P. aerugi- also has been shown to mediate cor- excess cholesterol in lysosomes, a key nosa senses changes in its environ- onary heart disease, or CHD, but so waste disposer in cells. While there ment and alters the expression of its far clinical trials of drugs that target are no approved drugs to treat NPC,

MAY 2020 ASBMB TODAY 17 JOURNAL NEWS

Shedding light on lipid raft formation

A major recent advance in our understanding of cellular mem- team from Stony Brook University led by Guangtao Li write that they branes is the discovery of so-called lipid rafts or nanodomains. These were able to circumvent this common shortcoming. The researchers domains are membrane regions enriched for certain lipids and proteins used a technique called Förster resonance energy transfer, which and are predicted to be important for a range of biological processes. sensitively can detect low nanometer distances between labeled lipids With many questions outstanding on their function, structure and otherwise undetectable by normal microscopy. With this technique, the formation, nanodomains are the subject of significant research in the researchers detected nanodomain formation in GPMVs at temperatures field of lipid membranes. at least as high as 38 degrees, where larger-scale separations disappear, Attempts to study these nanodomains, however, have been ham- confirming that nanodomains occur at physiological temperatures. Li pered by difficulties in detection, leading many studies to use model and colleagues also were able to modulate the lipid and cholesterol membranes called giant plasma membrane vesicles, or GPMVs, at compositions of their GPMVs, confirming that differing membrane low temperatures (0°C to 20°C). These conditions induce larger-scale compositions could influence domain formation. membrane ordering that can be detected by basic light microscopy With the importance of nanodomains in normal cellular function, techniques. While such studies have been valuable, researchers remain the ability to study these domains under more biologically relevant con- concerned that low temperature studies on these large-scale phase ditions is invaluable. Future research not only could expand our scientif- separations do not represent accurately the behavior of smaller nano- ic knowledge of nanodomain behavior but potentially allow scientists to domains in human cells under physiological conditions. target and manipulate these domains for new drugs and treatments. In a recent paper published in the Journal of Lipid Research, a DOI: 10.1194/jlr.RA119000565 — Kian Kamgar–Parsi

Lipid bilayers often are separated into disordered regions (1) and ordered nanodomains (2), with these relatively poorly understood nanodomains

ARTUR JAN FIJAŁKOWSKI/WIKIMEDIA COMMONS ARTUR JAN FIJAŁKOWSKI/WIKIMEDIA serving important biological functions.

2-hydroxypropyl-beta-cyclodextrin, with CD. occurred. These findings provide a or CD, is a promising molecule In a paper published in the Jour- more complete picture of CD’s role currently in human trials. However, nal of Lipid Research, Feltes and in mediating cholesterol homeostasis researchers know little about the her colleagues were able to confirm in NPC and could prove useful in details of CD’s mechanism of action. CD-dependent decrease in lysosomal CD’s continued development as a Previous studies have shown CD to cholesterol levels. Rather than re- drug for this disease. decrease lysosomal cholesterol ac- moving this cholesterol from the cell, cumulation but also have shown no however, CD promoted transfer of Using a fusion protein effect on total cellular cholesterol. To lysosomal cholesterol to the plasma to make fat levels fall reconcile these seemingly disparate membrane. From there, cholesterol Lipoprotein lipase, or LPL, is an findings, McKenna Feltes and a team either would exchange with lipopro- enzyme that metabolizes circulating from Washington University labeled tein-bound cholesterol or be routed triglycerides. When LPL activity is cholesterol and tracked its movement to the endoplasmic reticulum for compromised — as is the case in through model NPC cells treated processing if excess accumulation familial chylomicronemia syndrome,

18 ASBMB TODAY MAY 2020 JOURNAL NEWS

Identification of another risk factor for AITDs

Autoimmune thyroid diseases, or AITDs, are caused by a self-me- showed that HT is linked to a decrease of antennary alpha1,2 fucose diated attack on thyroid cells and ultimately affect thyroid gland func- and that enrichment of IgG N-glycan traits is associated with genes tion. Two common AITDs are Hashmito’s thyroiditis, or HT, and Graves’ FUT8 and IKZF1, which are essential for IgG core fucose formation. The disease. Before clinical symptoms appear, the human body produces researchers could not determine genetic variances between AITD and autoantibodies against the thyroid cells, which allows clinicians to IgG N-glycan traits; however, they showed that decreased core fuco- detect these illnesses. Researchers hypothesize that thyroid peroxidase sylation and antennary alpha1,2 fucose are not associated with gene antibody, or TPOab, a commonly used diagnostic marker, has a role in expression alteration in peripheral blood mononuclear cells. autoimmunity by modulating IgG glycosylation. In conclusion, the researchers propose a model in which FUT8 and Recently, Tiphaine C. Martin of King’s College and an international IKZF1 have an aberrant expression in a tissue-specific manner instead team published a paper in the journal Molecular & Cellular Proteomics of in the blood. They think this model will show that the altered expres- that focused on determining whether immunoglobulin G, or IgG, glyco- sion causes antibody-dependent cell-mediated cytotoxicity directed sylation is associated with TPOAb that is positive for AITD and whether against the thyrocyte due to afucosylated TPOAb antibodies. In turn, AITD and glycan structures share any genetic, heritable factors. they propose that this cytoxicity leads to AITD development. In this study, the researchers found that TPOAb level and AITD DOI: 10.1074/mcp.RA119.001860 are associated with decreased IgG core fucosylation. Additionally, they — Latavia Hill TIPHAINE MARTIN ET AL./MCP

This proposed model illustrates the role of FUT8 and IKZF1 in the development of autoimmune thyroid diseases.

MAY 2020 ASBMB TODAY 19 JOURNAL NEWS

Knocking out Nogo to rouse regeneration

The nervous system has the remarkable ability to adapt and cellular communication, exerting effects on other cells. So, could exo- respond to a variety of environmental signals. Its two components are somes be suppressing axon regeneration after traumatic injury? Yuichi the central nervous system and peripheral nervous system, or CNS and Sekine and colleagues at Yale University School of Medicine tested this PNS, respectively. The peripheral nervous system, which consists of the hypothesis by first expressing Nogo-A in a human cell line. They found nerves and ganglia outside of the brain and spinal cord, can heal after that the enzyme beta-secretase 1 created a fragment of Nogo-A, called a traumatic injury through spontaneous axonal regeneration. Curiously, Nogo-24, that escapes from cells into the surrounding fluid on the outer this same type of self-repair does not occur after CNS injury such as surface of exosomes. Then, to confirm that Nogo-24 inhibits axon regen- stroke or spinal cord injury. In 2012, Feras Akbik of the Yale School eration when presented to cells on the surface of exosomes, the authors of Medicine and a team of researchers explored this disparity and cultured neurons and observed that injured axons regrow in the absence identified Nogo-A, a protein expressed by neurons and nonneuronal of exosomes with the Nogo-24 fragment. Finally, the researchers found cells called oligodendrocytes, as a molecule that hinders nerve cell that the Nogo-24 was present in an exosomal fraction of mouse spinal regeneration. However, how Nogo-A might disrupt axonal regeneration cord tissue that was injured but not in tissue from uninjured animals. after spinal cord injury remained a mystery. In a recent paper published Their findings suggest a new pathway by which disruptive signals in the Journal of Biological Chemistry, researchers describe how may be presented to prevent axon regeneration after injury. they helped to solve this mystery by identifying Nogo-A in a different DOI: 10.1074/jbc.RA119.009896 location: exosomes. —Anand Rao Exosomes are small cellular storage compartments that can traffick select cargoes throughout cells, and their contents can mediate SIENNA DRAKE AND ALYSON FOURNIER SIENNA DRAKE AND ALYSON

This illustration shows how exosomal Nogo-24 may be released after spinal cord injury and signal through NgR1 to inhibit neuron regeneration.

20 ASBMB TODAY MAY 2020 JOURNAL NEWS

or FCS — plasma triglyceride levels amino acids, such as leucine and me- gen-dependent manner. are elevated dangerously, and few thionine, signal mTORC1 through DOI: 10.1074/jbc.RA119.010315 clinical options are available to cor- the well-characterized Rag guanosine rect them. triphosphatase signaling pathway, An essential protein In a recent paper published in the while others, such as glutamine and in the Journal of Biological Chemistry, asparagine, signal through Rag-inde- Each day, millions of cells un- Amitabh Nimonkar and colleagues pendent pathways. Their work was dergo cell division to make daughter at Novartis Institutes for BioMed- published recently in the Journal of cells. This extensively studied cell ical Research showed that a fusion Biological Chemistry and shows cycle consists of four phases: a gap complex between LPL and the LPL that mTORC1 is differentially phase, synthesis, a second gap and transporter glycosylphosphatidy- regulated by amino acids and may be . Each checkpoint in this linositol-anchored high-density useful in furthering treatments for cycle is crucial, because any irregu- lipoprotein-binding protein 1, or metabolic disease. larities can lead to uncontrollable cell GPIHBP1, demonstrates high enzy- DOI: 10.1074/jbc.AC119.011578 growth, also known as cancer. matic activity and is resistant to in- A recent paper by Patrick Herr hibitors of LPL. Moreover, injection Enzyme regulates of Karolinska Insitutet and a team of the LPL–GPIHBP1 fusion protein ’s role in cells in Sweden, published in the journal lowered triglyceride levels in several The post-translational modifica- Molecular & Cellular Proteomics, mouse strains, indicating its thera- tion ubiquitination plays an import- focused on dissecting cell cycle dy- peutic potential for the treatment of ant role in many cellular processes, namics at the protein level in asyn- FCS and other triglyceride-related including cellular immunity, tran- chronous cells, using methods other diseases. scriptional regulation and membrane than chemical synchronization. The DOI: 10.1074/jbc.RA119.011079 protein maintenance. Deubiquitinas- researchers characterized protein os- es, or DUBs, such as Cezanne — an cillation patterns over the course of Amino acids activate enzyme that cleaves ubiquitin from the cell cycle and found that many mTORC protein substrates — are crucial for vital processes are affected. The The mammalian target for rapa- the regulation of ubiquitin signaling. team detected differences in mRNA mycin 1, or mTORC1, senses amino In a recent paper published in the abundance and phosphorylation pat- acids in the cellular environment and Journal of Biological Chemistry, terns among cell cycle phase groups. regulates cell growth and metabolism Julia Mader and colleagues at Goethe They characterized predicted cell to match nutrient availability. Dis- University in Frankfurt, Germany, cycle–dependent proteins, including ruption in this sensing mechanism discovered that adding hydrox- S-adenosylmethionine synthase, or can result in metabolic disease and yl groups to asparagine regulates MAT2A. They found that MAT2A cell growth disorders. Cezanne’s interaction with ubiquitin nuclear localization was enriched in Using cultured cells, immuno- in an oxygen-dependent manner. synthesis and in the second gap and blotting, immunofluorescence and These findings demonstrate that mitosis, and they speculate that this pharmacological approaches, Delong Cezanne-mediated removal of ubiq- protein is essential for epigenetic Meng of the University of Texas uitin and the related physiological histone methylation during DNA Southwestern Medical Center and processes affected by the removal of replication. colleagues demonstrated that some ubiquitin are regulated in an oxy- DOI: 10.1074/mcp.RA120.001938

Latavia Hill (lataviahill@ Kian Kamgar–Parsi Anand Rao ku.edu) is a graduate ([email protected]) ([email protected]) is a student studying studied biophysics at the science communicator microbiology at the University of Michigan for the ASBMB. Follow University of Kansas. and is a consultant for the him on Twitter @ pharmaceutical industry. AnandRaoPhD.

MAY 2020 ASBMB TODAY 21 FEATURE

A LEGACY OF TYROSINE

Tony Hunter, the Salk Institute ony Hunter hasn’t had a and biochemistry, which he went biochemist who discovered this clean shave in more than on to study as an undergraduate at 40 years. Christ’s College at the University of amino acid’s phosphorylation, “On one of his trips Cambridge. was scheduled to speak about to the Grand Canyon, he “We had a great set of teach- stopped shaving, and he ers, including Nobel Prize winners his work at EB 2020. Thasn’t shaved since,” said Walter Eck- like Fred Sanger and Max Perutz,” hart, Hunter’s former postdoctoral Hunter said of his time at Christ’s. “I advisor and longtime colleague at the didn’t really know that I wanted to By John Arnst Salk Institute for Biological Studies. go into research as a career, but they “Well, he’s trimmed his beard.” were a major influence.” A member of the National After Hunter graduated, he Academy of Science since 1998, stayed at Cambridge and began Hunter has received many awards graduate studies with the molecular and honors throughout his career, biologist Asher Korner in the depart- including the 2018 in ment of biochemistry, obtaining his biopharmaceutical science; he was Ph.D. in 1969. scheduled to deliver the 2020 Tang After a brief fellowship at Prize Award Lecture in April at Christ’s, Hunter followed his first Experimental Biology in San Diego wife, Philippa “Pippa” Marrack, before the meeting was cancelled due whom he had met while they were to the COVID-19 pandemic. His both graduate students, to California discovery that the activity of tyrosine for her postdoctoral fellowship at the kinases drives the growth of cancer- University of California, San Diego. ous cells ultimately led to the devel- “I tagged along and ended up opment of tyrosine kinase inhibitors with the Salk because her advisor, and the groundbreaking leukemia Alan Munro, had done a short sab- therapy Gleevec, as well as an entire batical here to learn immunology, so subfield of biochemical regulation. he suggested that I come.” There, for the first time, he An English upbringing encountered tumor — and an Born in 1943, Anthony Rex abundance of sunshine. Hunter grew up in the southeast of “I thought when I left to do my England near Canterbury. He at- postdoc, that I’d be here two years, tended the venerable Felsted School find out about California and then in Essex, which celebrated its 450th go back to Cambridge to the rest of anniversary in 2007. During his time my career, but it didn’t work out that there, he gravitated toward biology way,” he said. “It was a huge culture

22 ASBMB TODAY MAY 2020 FEATURE COURTESY OF TONY HUNTER COURTESY OF TONY

Tony Hunter, who discovered tyrosine phosphorylation in 1979, served as the director of the Salk Institute Cancer Center from 2008 to 2016.

shock, obviously.” protein of a second oncovirus, has kinase. This led Hunter to generate After a brief return to Cam- kinase activity. According to Hunter, sequence alignments of the viral bridge, Hunter settled at the Salk this provided an immediate hint that tyrosine kinase proteins, as well as Institute in 1975. Four years later, human cells might also have tyrosine protein kinases that phosphorylate he made his crucial discovery that kinases. serine and threonine, which revealed unchecked tyrosine phosphorylation “We found, in short order, that a that the catalytic domain of tyrosine in cancerous cells was behind their Rous virus encodes a similar kinase has a series of conserved short aberrant growth. tyrosine kinase activity,” Hunter said. sequence motifs that are essential for “And since that gene was derived transferring phosphate. Tyrosine and tumors from the cellular SRC gene by viral He and various colleagues then By Hunter’s recollection, he and capture, we already knew that cells used PCR amplification and degen- his colleagues stumbled on tyrosine must have at least one enzyme with erate oligonucleotide probes that phosphorylation through their work this type of activity. We realized that recognize the short sequence motifs on the polyoma DNA tumor virus. having two viruses using a similar to identify new tyrosine and serine In a 1979 paper in the journal Cell, mechanism could mean that it was a kinase genes. When those genes were they described their findings that common mechanism of viral trans- combined with analysis of the human phosphate in cell culture was associat- formation, but we didn’t know at the genome sequence, the researchers ed with one of the three proteins the time it would be important in cancer ultimately wound up discovering virus encodes to transform normal in the way that we now know.” nearly 500 human kinases, about 90 cells into tumor cells. They quickly Not long afterward, a group of which are tyrosine kinases. followed it with another paper in the led by Stanley Cohen at Vanderbilt According to Hunter, the first Proceedings of the National Academy University reported that the trans- major hints that tyrosine phosphor- of Sciences wherein they reported membrane epidermal growth receptor ylation played an important role in finding that v-SRC, the transforming protein, or EGFR, was also a tyrosine growth of cancer cells came when

MAY 2020 ASBMB TODAY 23 FEATURE

scientists at the National Cancer In- translocation, which occurs in 95% Hunter said. “So much so that some stitute and Erasmus University found of CML cases, would shut down of the patients who went on the that chronic myeloid leukemia, or cancerous CML cells. In the early drug experimentally in 1999 and CML, was caused by a chromosomal 1990s, Brian Druker, a physician sci- 2000 when it was in trials are still on translocation between chromosomes entist at Health and Science the drug 20 years later because it’s 9 and 22 that fuses together the University who had been working well-tolerated and is keeping their kinase-producing ABL gene with the with CML patients, reached out to disease in check.” BCR gene. They reported their find- scientists led by Nicholas Lyndon at Today, 40 tyrosine kinase inhibi- ings in three papers that appeared in the Swiss pharmaceutical company tors have been approved by the Food 1982 and 1983 in the journal Nature Ciba-Geigy, which later became No- and Drug Administration as cancer and in 1984 in the journal Cell. vartis, who were developing tyrosine therapies. “The fusion gene encodes a kinase–blocking compounds. After “And kinase inhibitors as a class chimeric BCR-ABL protein that is homing in on the kinase inhibitor are now up over 65,” Hunter said. constitutively activated as a tyrosine STI-571, they found both that it “That’s pretty impressive, considering kinase,” Hunter said. “Within a few caused a 92% to 98% decrease in this all started only about 20 years years, several other tyrosine kinase the number of cells exhibiting the ago in a huge effort in this area, and genes were shown to be mutationally BCR-ABL fusion in human bone there are many, many more in clin- activated or overexpressed in human marrow cells predisposed to develop ical trials. I’m sure the number will cancer.” into leukemia cells and that it kept continue to increase.” Work by and by the cancerous cells from returning. Owen Witte’s group at the Univer- Within short order, the drug, which Knowledge and notes sity of California at Los Angeles had was renamed imatinib and later While the Salk Institute was laid the groundwork for the hypoth- marketed as Gleevec, entered human founded in 1960 as a place where esis that inhibiting the activity of the trials. scientists could devote themselves to BCR-ABL kinase made by the gene “It proved to be a very effective research free of teaching obligations, fusion created by the Philadelphia treatment for early-stage CML,” mentoring remains a significant part of each Salk scientist’s career. Ruth Palmer, a biochemist at the University of Gothenburg in Sweden who was a postdoctoral fellow in Hunter’s lab from 1996 to 2000, recalled the latitude Hunter gave his

COURTESY OF RUTH PALMER postdocs and his openness to their ownership of projects. “He’s very generous in terms of whatever you generate in his lab; he supports it and then you can take it with you completely,” Palmer said. “For me, that was incredibly im- portant. When I started my own lab after that, which I did here, I felt it was actually quite easy because I had already become very independent in his lab.” An avid traveler, Hunter attends many conferences each year. In 2017, he met up with his former Jonathon Pines, now head of postdoctoral researchers Clare Isacke (left) and Ruth Palmer (right) at the European Molecular Biology the Division of Cancer Biology at Organization’s annual meeting, during which the two were inducted into the organization. the Institute for in

24 ASBMB TODAY MAY 2020 FEATURE IMAGES COURTESY OF TONY HUNTER COURTESY OF TONY IMAGES

Tony Hunter has rafted the Grand Canyon several times, most recently in 2010. Top left: Marble Falls Canyon, a launching point on the Colorado River north of the Grand Canyon, in 1981. Top right: Lava Falls Rapid in 1987. Bottom: Hermit Rapid in 2010.

London, was a postdoc of Hunter’s within a very short period of time, of the literature meant that he’s also from 1988 to 1992. he knew as much about the cycle able to make connections that might “Tony was incredibly generous,” as I did. He has this extraordinary not be apparent to other people. He’s Pines said. “When I arrived, there capacity to absorb and remember always been able not only to master wasn’t anybody working on the cell information.” a particular subfield, but also make cycle in his lab, but he was he was During his time in Hunter’s connections between fields.” interested in cell cycle regulation.” lab, Pines cloned the first human According to Pines, and every During his doctoral work at Cam- , A and B1, and mapped their other scientist interviewed for this bridge with , who had interactions with proteins involved in feature, this knowledge takes a worked alongside Hunter when they promoting cancer and regulating the physical manifestation in decades of were both graduate students, Pines cell cycle. voluminous, detailed notes. had cloned B, a protein critical “I learnt a huge amount from “Tony’s notes are famous in the for regulating mitosis in animal cells. him in terms of how to design re- lab,” Pines said. “He listens to every “So he just said, ‘Why don’t search that was productive, partic- talk and writes everything down in you go ahead and continue to work ularly how to interpret data,” Pines pencil, because it’s fireproof.” on the cell cycle,’” Pines said. “And said. “His encyclopedic knowledge During Hunter’s second stint at

MAY 2020 ASBMB TODAY 25 FEATURE IMAGE COURTESY OF TONY HUNTER COURTESY OF TONY IMAGE

When Tony Hunter returned to La Jolla in 1975, he bought a Volkswagen Beetle, which many Salk faculty members have borrowed over the years. In 2003, the Beetle hit 240,000 miles, the equivalent of a one-way trip to the moon.

Cambridge as a postdoc, a fire that Tony remembered that this person’s Since 1972, Hunter has gone on started on a Friday night destroyed father had also been a job candidate 35 rafting trips in 10 rivers in Idaho, the lab he was working in and some years before,” Eckhart said. “So Washington, Oregon, California, burned much of the adjoining room he took great pleasure in showing Utah and Arizona, with many early where the scientist kept their notes. this guy the notes from his father’s members of his lab tagging along at Despite being burned and black- seminar.” least once. ened, however, Hunter’s notebooks “We went down a river called remained legible and salvageable. Beyond the lab the Selway, and Tony had one of “It turns out you can see pencil During his time in southern those big inflatable rafts that take writing even if the pages are totally California, Hunter has taken an ad- about six people,” Pines said. “He black because it’s a slightly different venturer’s spirit to the western states, navigated the class IV and V rapids color,” Hunter said. Although inves- hiking in the deserts, camping on with aplomb.” (Rapids are graded on tigators could not determine the ori- deserted beaches, off-roading on the a scale of I, which are swimmable, gin of the fire, Hunter believes it was Baja peninsula and rafting rivers with to VI, which are off limits and can caused by ether, which is extremely all manner of rapids. be life-threatening even to expert flammable. “I’ve enjoyed all the outdoor rafters.) Moreover, according to Hunt- activities that are possible in Califor- Many of Hunter’s trips have been er’s longtime colleague Eckhart, the nia that are not available so easily in on the Colorado River, through the bearded biochemist can recall his Europe,” he said. “I still enjoy doing Grand Canyon. notes on a whim. some of those things, although phys- “I think the ultimate trip is “We had a job candidate at the ically they’re not quite that easy to do rafting through the Grand Can- Salk last year, and it turns out that anymore.” yon,” Hunter said. “It’s a 230-mile

26 ASBMB TODAY MAY 2020 FEATURE

trip, and the maximum amount of to analyze what he can from home, which could then be used as a new time you can take, according to the and he hosts journal club meetings therapeutic approach for pancreatic permit, is 18 days. In the days before with Salk cancer researchers on Zoom cancer, a topic Hunter has been pur- they instituted that limit, our longest every Friday morning. suing in his own lab in recent years. trip was 28 days. I calculated that I Hunter and his wife, Jennifer Hunter remains optimistic about have spent nearly six months at the Price, have two sons; the young- the future of his research, which in bottom of the Grand Canyon over er, James, born in 1996, recently recent years has begun to focus on the years.” graduated from Virginia Tech with both pancreatic cancer and histidine a degree in mechanical engineering kinases. Pancreatic cancer and their older son Sean, born in “Right now, we think it’s still a and histidine horizons 1990, recently defended his thesis at little unclear what histidine phos- Though his laboratory, along with Stanford University. His project was phorylation does,” he said. “In some all others at the institute, is closed to engineer a fragment of the leuke- cases, it’s an enzyme intermediate, during California’s COVID-19 shel- mia inhibitory factor receptor to act but in the other cases, it has regulato- ter-in-place order, Hunter continues as a ligand trap for the LIF cytokine, ry effects … like any new phosphory- lation, you have to prioritize what to study, because it takes a lot of effort just to analyze one site. So we are trying to do that. “I just got a seven-year outstand- ing investigator award from the NIH to study the role of histidine phos- phorylation in cancer. So, I’ll be busy for a while longer.”

Author’s note: Geoff Wahl and Ron Evans contributed information about Tony Hunter for this article. HUNTER COURTESY OF TONY IMAGE

John Arnst (jarnst@ asbmb.org) is an ASBMB Today science writer. Follow him on Twitter @arnstjohn.

In October 2018, Hunter dyed his beard purple for the Purple Stride San Diego walk, which raises funds for the Pancreatic Cancer Action Network.

MAY 2020 ASBMB TODAY 27 Get a decision before your flies emerge.

At 18°C, Drosophila take 19 days to eclose. JBC takes only 17 days on average to reach first decision about your paper. (Don’t worry, we don’t make our reviewers work at 18°C.)

Learn more about fast, rigorous review at jbc.org.

www.jbc.org The COVID-19 issue

30 A small army of researchers races to build a coronavirus interactome By John Arnst

32 Could an old malaria drug help fight SARS-COV-2? By John Arnst

34 Anatomy of a molecule: what makes remdesivir promising? By Laurel Oldach

36 Slipping past the proofreader By Laurel Oldach

44 Scientist uses community organizing skills to mobilize researchers against COVID-19 By Laurel Oldach

46 Researchers retool genomics labs to provide virus testing By Laurel Oldach

48 “We are doers. We want to get involved.” By Laurel Oldach

MAY 2020 ASBMB TODAY 29 FEATURE

A SMALL ARMY OF RESEARCHERS RACES TO BUILD A CORONAVIRUS INTERACTOME By John Arnst

s institutions shuttered across the U.S. ahead of shel- Krogan conceived and coordinated the collaborative ter-in-place orders, Nevan Krogan’s lab at the Uni- effort, which was carried out by scientists from 22 labs at Aversity of California, San Francisco, raced to gather the Quantitative Biosciences Institute as well as from the the last bits of data they needed to assemble a protein University of Michigan, the University of California San interaction map to identify drugs that might work against Diego, the Icahn School of Medicine at Mount Sinai, the SARS-CoV-2, the virus that causes COVID-19. Howard Hughes Medical Institute, the Fred Hutchinson “We actually collected the last sample for the map Cancer Research Center, the European Molecular Biology about six hours before they shut everything down,” said Laboratory and the Institut Pasteur. When it came to Krogan, a professor in the department of cellular and determining which drug targets could interact with the molecular pharmacology at UCSF and the director of the viral proteins, Krogan looked to UCSF chemists Brian university’s Quantitative Biosciences Institute. “It was Shoichet and Kevan Shokat. really a race against the clock, and it was a testament to “For the last 10 years, we’ve been working on getting the scientists in the lab essentially working for days on end drugs for undruggable targets,” Shokat said. “I looked in without sleeping.” the map for weird targets that nobody thinks of as In a study uploaded to the preprint site bioRxiv last traditional drug targets.” week, Krogan and 93 co-authors list 69 drugs that might That meant the target list included not only the be effective in targeting COVID-19, the coronavirus that immunosuppressant rapamycin and the drugs chloro- is testing the limits of healthcare systems and safety nets quine and remdesivir, which are being evaluated against around the world. To identify these drugs, they built an SARS-CoV-2 in clinical trials around the world, but also interactome that showed how 332 human proteins inter- cancer drugs such as dabrafenib and natural products such acted with proteins from 26 of the virus’ 29 genes. as WDB002, a cyclic polyketide synthase, that are being “It’s a kind of a blueprint of how the virus comes evaluated for clinical use. in and hijacks and rewires the host during the course of All of those drugs are being evaluated against the infection,” Krogan said. “What happens is you put a hook coronavirus in the biosafety-level 3 labs at the Institut Pas- on each one of (the viral genes) and you express the genes teur in Paris. There, Marco Vignuzzi and his lab members, in human cells, and then what you do is pull out, fish out, including postdoctoral fellows Bjoern Meyer, Veronica the corresponding protein with that hook, and then you Rezelj and Cassandra Koh, must work one or two at a see by mass spectrometry which other human proteins time due to the city’s continuing lockdown. are physically interacting or physically talking to the viral “We are currently testing 20 compounds,” Meyer said. proteins.” “Overall, I think the whole panel will include something

30 ASBMB TODAY MAY 2020 FEATURE NIAID/RML

This scanning electron microscope image shows SARS-CoV-2 (round gold objects) emerging from the surface of cells cultured in the lab. The virus shown was isolated from a patient in the U.S. along the lines of 65 or 69 compounds.” Their efforts are it. But here you had two weeks,” he said. “It was really being duplicated, for the sake of corroboration, by Kris M. outstanding. You just wake up at 7, work till 11 and then White and Lisa Miorin at the Icahn School of Medicine at check your phone until it’s bedtime. It’s crazy, everybody’s Mount Sinai in New York. spirit has been amazing.” The collaboration includes researchers at UCSF who The scientists in San Francisco must work at a distance hadn’t previously brought their skills to bear on viral prob- from each other and from their labs, but their colleagues lems, such as structural biologist Natalia Jura. in New York and in Paris continue to evaluate compounds “We were just so motivated to contribute that every- against the coronavirus in their level 3 labs, the sec- body really put on hold what they were doing … and just ond-highest level of biosafety containment. As Krogan and started reading papers about and accumulat- colleagues whittle down their list of drugs, he hopes that ing expertise in this area,” Jura said. “As a structural biolo- even more scientists will take advantage of their interac- gist, I was most interested in understanding what is known tion map to find new leads about both SARS-CoV-2 and about how viral proteins interact with the host proteins.” drugs that might work against it. Most of the paper’s authors are now analyzing data “It’s a very rich data set, and they can make predictions remotely, with little access to their labs, according to about the biology of virus that we didn’t find, and they Krogan. can also make predictions about other drugs and com- “We’re having regular Zoom calls with scientists — I pounds,” he said. “What I’m hoping for here, the silver think last time there were 100 different scientists on the lining in all of this, is that we’re setting a new paradigm of Zoom call discussing this map,” he said. Krogan and how to do science. And hopefully, this infrastructure stays colleagues at the UCSF-affiliated nonprofit Gladstone in place so that we’re in a better position to tackle the next Institute previously assembled drug interaction maps pandemic.” for HIV and for the Ebola, Zika and dengue viruses with much longer timeframes. “Normally, that takes a few years,” he said. “Well, we expedited this for a couple of weeks. And that’s a testa- John Arnst ([email protected]) is an ment in my opinion to the collaborative effort that was ASBMB Today science writer. Follow him on underway.” Twitter @arnstjohn. Shokat is impressed by the speed with which the group was able to pull together its data. “Usually we have six months, nine months to do

MAY 2020 ASBMB TODAY 31 FEATURE

Could an old malaria drug help fight SARS-CoV-2? By John Arnst

hloroquine might be getting the bark of cinchona trees native to anti-inflammatory agent. They rec- new life as an antiviral treat- Peru and used for centuries to treat ommended that it could be used in C ment for the novel coronavirus malaria. Chloroquine was an essen- combination with compounds that that emerged in Wuhan, China in tial element of mass drug adminis- prevent replication. Nevertheless, in late 2019 and has since become a tration campaigns to combat malaria 2009, the Belgian group found that pandemic with more than 2.5 mil- throughout the second half of the lethal infections of human coro- lion cases worldwide. For decades, 20th century, and remains one of the navirus OC43, a relative of SARS, the drug was a front-line treatment World Health Organization’s essen- could be averted in newborn mice by and prophylactic for malaria. tial medicines. However, after the administering chloroquine through In a three-page paper published malaria parasites Plasmodium falci- the mother’s milk. in early February in Cell Research, parum and Plasmodium vivax began Chloroquine raises the pH in scientists at the Wuhan Institute of exhibiting resistance to the drug in host-cell lysosomes, which interferes ’s State Key Laboratory of the 1960s and 1980s, respectively, with viruses’ attempts to acidify the Virology write that both chloroquine it was replaced by similar antima- lysosomes, a prerequisite to forma- and the antiviral remdesivir were, larial compounds and combination tion of the autophagosomes that cells individually, “highly effective” at therapies. Chloroquine is still widely use to eat themselves. In the Cell inhibiting replication of the novel used against the three other species Research paper, the researchers found coronavirus in cell culture. Their of plasmodium and to treat auto- that the drug was effective at inhibit- drug screen evaluated five other immune disorders and some cases ing the virus as it was both entering drugs that were not effective. The of amebiasis, an intestinal infection and exiting cells. authors could not be reached for caused by the amoeba Entamoeba Craig Cameron is a virologist at comment. histolytica. the University of North Carolina at Though the paper is brief, John Chloroquine’s antiviral proper- Chapel Hill. “There is mounting ev- Lednicky, a professor at the Univer- ties were explored in the mid-1990s idence that many viruses hijack this sity of Florida’s Emerging Pathogens against HIV and in the following de- cellular autophagy pathway for the Institute, found its results intriguing. cade against severe acute respiratory good of the virus, but it is not com- “It’s interesting in that it really lacks syndrome, or SARS, which is closely pletely clear why,” he said. “From a lot of details but, nevertheless, if related to the novel coronavirus. In this, one can imagine chloroquine as you look at the data as presented, at 2004, researchers in Belgium found an antiviral.” least in vitro, it seems like chloro- that chloroquine inhibited replica- The second compound, remde- quine can be used as an early-stage tion of SARS in cell culture. The fol- sivir, is a nucleoside analog dis- drug,” he said. “It would be very lowing year, however, another team covered in 2016 that inhibits viral good if these types of experiments at Utah State University and the polymerase activity, shutting down were repeated by more laboratories Chinese University of Hong Kong transcription and synthesis of viral to see whether the same results occur evaluated a gamut of compounds RNA. This gives it antiviral activity across the board.” against SARS replication in mice against a broad range of - Chloroquine is a synthetic form infected with the virus, finding that es, including Ebola (for which the of quinine, a compound found in chloroquine was only effective as an drugmaker Gilead developed and

32 ASBMB TODAY MAY 2020 FEATURE YINAN CHEN/GOOD FREE PHOTOS

tested it, unsuccessfully, during the chloroquine he said, is at how many clinical trial by the French microbi- 2018-2020 epidemic in the Demo- days into an infection it can be effec- ologist Didier Raoult that caught the cratic Republic of the Congo) and tively administered to someone sick attention of President Trump. In late coronaviruses. with the new coronavirus. March, a man in Arizona died after “The fact that this drug works “As an analogy, Tamiflu works ingesting a form of chloroquine used against this virus is not unexpected, very well against susceptible to clean aquariums; according to his especially in vitro,” Cameron said. A virus strains as long as you take it wife, who also ingested the cleaner “Accumulation in the lungs to a level early enough,” he said. “And that’s and lived, they took it after watching that is effective is likely the bigger what we have to determine with chlo- a White House press conference in issue as a therapeutic for humans.” roquine, whether it can be used when which the president touted its effec- The Wuhan Institute of Virology somebody has been sick for more tiveness. Additionally, many people submitted a patent Jan. 21 for the than a few days. But the indication with lupus and arthritis who rely on use of remdesivir to fight the new so far, based on this paper and past hydroxychloroquine are now facing coronavirus in China; this may set up work with SARS, is that it might be a shortages of the drug. a battle with Gilead over intellectual useful drug.” As some doctors on the front property rights. In their filing, the in- However, both chloroquine and lines are using the drug — one stitute noted that they did not apply its less-toxic derivative hydroxychlo- of very few tools in their medical to patent chloroquine phosphate be- roquine, marketed as Plaquenil and arsenals against COVID-19 — for cause it has been marketed in China prescribed to people with autoim- patients at various stages of the dis- and has an extant supply chain. mune disorders including lupus, do ease, various clinical trials have been Lednicky is optimistic about carry risks of cardiomyopathy and launched to evaluate its effectiveness the prospects for treating the new heart failure in patients who are both against the virus. coronavirus with remdesivir and older and pre-disposed to cardiac chloroquine. disease.

“What’s important is that the Over the last two months, the John Arnst (jarnst@ selectivity index is relatively high for potential usefulness of chloroquine asbmb.org) is an both of them,” Lednicky said. “In and its less-toxic derivative hy- ASBMB Today science writer. Follow other words, they’re not expected to droxychloroquine as treatments for him on Twitter @ have a lot of side effects.” COVID-19 has become a highly arnstjohn. The biggest question regarding politicized subject following a flawed

MAY 2020 ASBMB TODAY 33 FEATURE

Anatomy of a molecule: what makes remdesivir promising? Experts weigh in on the chemistry of the potential SARS-nCoV-2 antiviral

By Laurel Oldach

he World Health Organization in late January The compound and ATP have some important differ- convened experts to discuss experimental ences, but some features are very similar. ASBMB Today therapeutics for patients with the emerging spoke to medicinal chemist Katherine Seley–Radtke at the T coronavirus with no name, no vaccine and no University of Maryland, Baltimore County, and structur- treatment. The panel reported that “among the different al virologist Craig Cameron at the University of North therapeutic options, remdesivir was considered the most Carolina, Chapel Hill about what makes the molecule promising candidate.” interesting. Within weeks, a clinical trial of the compound was underway in China. Results are expected in April; in the meantime, the outbreak of SARS-nCoV-2, the virus that causes COVID-19, has become a global pandemic. Remdesivir is a nucleotide analog, one of the oldest classes of antiviral drugs. It works by blocking the RNA polymerase that coronaviruses and related RNA viruses need to replicate their and proliferate in the host body. The molecule originally was synthesized as part of a screen for inhibitors of the virus RNA polymerase. Its inventors at Gilead Sciences decided to move forward with a different nucleotide analog compound to treat hepatitis C. But RNA-dependent RNA polymerases are conserved between many viruses. Experiments in vitro, in cell culture and in animal models have shown that remdesivir has broad-spectrum activity against RNA viruses, including filoviruses (like the one that causes Ebola) and coronaviruses. Remdesivir resembles the RNA base adenosine, shown here as a monophosphate.

My whole career has had the RNA “ dependent RNA polymerase at the center of our studies, because it really is a very well validated drug target. –Craig Cameron ” Craig Cameron Katherine Seley-Radtke

34 ASBMB TODAY MAY 2020 FEATURE

3. C-nucleoside bond Usually, the glycosidic bond connects the 1’ carbon in the ribose ring to a nitrogen 5. Phosphate in the base. But in remdesivir (and some other nucleoside analogs) the sugar and “You see all that flotsam and jetsam the nucleobase are connected by a bond coming off at the 5’ hydroxy?” said Kath- between two carbons. erine Seley–Radtke. Among medicinal 1. 3’ hydroxy group “It definitely provides much greater chemists, this type of protecting group is Different classes of nucleotide analogs stability (against) nucleases and other en- casually known as “a McGuigan ProTide.” have different effects on polymerases. zymes that can cleave the nucleobase from Designed by medicinal chemist Chris Mc- Remdesivir is in a class called nonobligate the sugar,” said Katherine Seley–Radtke, a Guigan in the 1990s, this type of protecting chain terminators, due to the presence of medicinal chemist at the University of Mary- group and its variations are widely used to the hydroxy group at carbon 3 in the sugar. land, Baltimore County who works on the deliver nucleotide analogs into cells. it should, in theory, be possible to add design and synthesis of antiviral nucleotide “It is a brilliant system, because it more nucleotides to a strand of RNA after analogues. With a C-nucleoside, “you’d have accomplishes two things,” Seley–Radtke remdesivir has been added. to break a carbon–carbon bond; whereas said. “No. 1, an issue with nucleosides is “That hydroxy group is what is required for in a normal nucleoside you’re breaking a that they’re polar and their phosphates continued synthesis of nucleic acid, whether it hemi-aminal bond, which is far less stable. are even more polar.” Masking the highly be RNA or DNA,” said virologist Craig Cameron, So having that carbon–carbon bond is a negative phosphate groups with esters or who studies the interactions between between great advantage.” amides reduces the molecule’s overall po- nucleoside and nucleotide analogs and viral larity, letting it cross the plasma membrane polymerases.. into cells. Recent research suggests that 4. 1’ cyano group Second, to be recognized by poly- remdesivir, when mixed with coronaviruses merases, the analog needs to resemble a RNA polymerases in vitro, doesn’t terminate Ask a group of chemists what jumps normal nucleotide triphosphate — which the synthesis of a new RNA strand right out at them about remdesivir, and most will means it needs to be phosphorylated. away. Instead, Cameron said, “it takes a few start with this dramatic feature. Substitu- “The first phosphorylation, either cycles between nucleoside and nucleotide tion at this carbon is unusual and probably by cellular or viral kinases, is oftentimes analogs and viral polymerases before you possible only because of the strength of the very difficult,” Seley–Radtke said. “A lot can see the termination effect.” C-nucleoside bond. of those kinases are very, very picky in Those additional nucleotides may According to an article in the Journal terms of recognition.” By arriving into the shield remdesivir from coronavirus of Medicinal Chemistry, the cyano group cell with its first phosphate already in proofreading enzymes that remove was initially added because a precursor tow, remdesivir and other nucleoside and unnatural nucleotide analogs. molecule, monophosphate inhibitor of viral nucleotide analogs and viral polymerases RNA polymerases, also blocked the mito- skip that rate-limiting step. After the pro- 2. Base pairing to uracil chondrial RNA polymerase in mice. To make tecting groups are cleaved, the nucleotide a molecule without such toxic side effects, analog is a reasonable substrate for later In adenosine in double-stranded RNA, chemists at Gilead tried a series of substi- nucleotide kinases. this face of the molecule is involved in base tutions at the 1’ carbon. The compound pairing with uracil. The two nitrogens act as with the cyano group worked best: it still

proton donor and acceptor, respectively, for blocked the hepatitis C polymerase but was Laurel Oldach (loldach@ hydrogen bonds to atoms in the uracil base. not incorporated by host-cell polymerases. asbmb.org) is a science Chemists think that remdesivir, by “You can’t predict activity. You have writer for the ASBMB. Follow her on Twitter @LaurelOld. presenting a very similar binding face, gets to make it and test it,” Seley–Radtke said. incorporated into a growing RNA strand by “But even small changes can have amazing viral polymerases. consequences.” MAY 2020 ASBMB TODAY 35 FEATURE

n the early 2000s, when experts believed human coronavirus infection caused noth- SLIPPING PAST THE ing worse than the , Mark IDenison struggled to land adequate federal PROOFREADER grants to support his lab at the Vanderbilt University Medical Center. The virus family that causes A virologist and clinician, Denison had been studying coronaviruses since 1984, when SARS, MERS and COVID-19 only two of the seven coronaviruses current- has an unusual ability to evade ly known to cause disease in humans were known. Those two, along with several other treatment. Can new drug coronaviruses, cause colds. candidates overcome it?

By Laurel Oldach VANDERBILT UNIVERSITY MEDICAL CENTER VANDERBILT

Mark Denison, a professor at Vanderbilt University Medical Center, has studied coronaviruses since the 1980s.

Denison initially found the virus he studied, which affects only mice, interesting because it leads to a mouse version of multiple sclerosis. Along the way, he became interested in how the virus replicated — but persuad- ing funders to support his work was a real challenge. In early 2003, he and his wife Laura were on vacation in Florida having a difficult Vials containing a solution of the nucleotide analog remdesivir conversation. “I think the work is important. are capped at a Gilead production facility. I think the models are important,” he recalled saying. “(But) I don’t know if I can maintain a career.” That was the day he learned about the pathogen behind the deadly respiratory dis-

GILEAD SCIENCES ease SARS, which had been making headlines.

36 ASBMB TODAY MAY 2020 FEATURE

“I’m literally on the beach with my wife, and someone came down from the hotel and told me that I had a phone call,” he said. The call was from a colleague, sharing the news that the pathogen responsible for the disease had been identified, and it was a coronavirus. Severe acute respiratory syndrome swept from southern China into 26 countries in 2002 and 2003, killing about 10% of the roughly 8,000 people it infected. Although the outbreak was contained, coronaviruses were recognized suddenly as a serious prob- lem of potentially pandemic proportion. Almost two decades later, the work done in Denison’s lab has turned out to be instru- mental in developing a molecule, called rem- Craig Cameron, a virologist at the University of North desivir, that entered large-scale clinical trials Carolina at Chapel Hill, studies RdRp molecular mechanisms just weeks after SARS-CoV-2, the coronavirus in picornaviruses. that causes COVID-19, was identified. The molecule, it seems, can overcome called ribavirin. At the time, ribavirin was the coronaviruses’ superpower: their ge- first-line drug for many RNA viruses. nome-proofreading ability. This ability isn’t Ribavirin targets a called the found in other RNA viruses, and it makes RNA-dependent RNA polymerase, or RdRp, coronaviruses resistant to a majority of drugs which is responsible for replicating the coro- used against other RNA viruses. navirus genome. When ASBMB Today went to print in Craig Cameron is a virologist at the Uni- mid-April, researchers expected to learn in the versity of North Carolina at Chapel Hill who next few weeks whether remdesivir is an effec- studies RdRp molecular mechanisms in pi- tive treatment for patients with COVID-19. cornaviruses. “(RdRp) is a very well-validated In the meantime, Denison’s lab has contin- drug target,” Cameron said. “And it is one of ued to work to identify other molecules that those targets that actually have the potential might bypass viral resistance. of having pan-viral antiviral activity.” “This is a really key question for us: Is Ribavirin belongs to a class of antiviral this a new class of drug that can allow us to drugs called nucleotide or nucleoside analogs. better design more drugs that can bypass the Exactly how ribavirin works seems to vary proofreading function and inhibit the virus?” from one virus to another, making it a good said Denison, who now directs the pediatric illustration of the many possible modes of infectious diseases division at Vanderbilt. action of nucleotide analogs. Some work by blocking the viral polymerase, terminating a Not your average RNA viruses growing strand of RNA. Some are mutagens, Like many viruses that cause human dis- which slip into the growing viral genome, let- ease, coronaviruses have a genome composed ting the polymerase continue, but introduce of RNA. some molecular ambiguity to the next round When the virus behind SARS was iden- of replication that causes a cascade of errors in tified, one of the many experimental treat- later generations. Some block metabolic en- ments clinicians tried using was a molecule zymes, preventing the synthesis or processing

MAY 2020 ASBMB TODAY 37 FEATURE

of real ribonucleotides and thereby slowing “We were surprised that ribavirin was down replication. pretty toxic and not very effective (against) A systematic review of data from 30 clini- coronaviruses,” Canard said. cal trials, conducted after the SARS epidemic, Both the CNRS team in Marseille and showed no conclusive benefit of ribavirin Denison’s group in Tennessee set out to — or any other treatment that was tested — understand more about the virus that caused and some evidence that ribavirin had done SARS, and one of their major questions was patients harm. why ribavirin, broadly effective against other Ribavirin was the first example. Since RNA viruses, had failed against this one. then, most nucleotide analogs tried against The answer lay in the virus family’s large coronaviruses that cause SARS and Middle genome and how it evolved to protect itself. East respiratory syndrome, or MERS, have “The reason (RNA viruses) are thought to not been effective treatments. Referring to be so successful is because their polymerases ribavirin and the classic nucleoside analog make mistakes,” Denison said. “They lack the 5-fluorouracil, which works as a mutagen, ability to correct mistakes, so they generate Denison said, “Coronaviruses are completely, mutant swarms of viruses that are ready for adaptation in different environments.” Structural biologists describe RNA-de- pendent RNA polymerases as resembling a cupped hand, with the fingers and thumb protecting the enzyme’s active site. As each new nucleobase in the template strand enters the active site, the polymerase coordinates a new incoming ribonucleotide by matching it against its counterpart in the existing strand. If the fit is right, the enzyme catalyzes a bond formation in the RNA backbone. If the fit is close enough, many viral RNA polymerases will catalyze a bond anyway. An error rate that can be as high as one mistake per 10,000 bases lets those mutant swarms arise. But for coronaviruses, the replication error rate is Bruno Canard started new research programs at the lower. French National Centre for Scientific Research after the Coronaviruses have some of the longest SARS outbreak. genomes in the RNA viral world. Whereas their closest cousins have genomes averaging entirely resistant to those drugs. You can soak 10 kilobases, coronavirus genomes are three them in it, and they have no effect.” times as long. With so much genetic material to copy, if coronaviruses mutated at the same Vigorous viral proofreading rate as other RNA viruses, they would accu- Bruno Canard is the principal investigator mulate so many mutations that they would of a group at the French Na- barely produce any viable progeny. tional Centre for Scientific Research. Before As researchers in the field came to un- the SARS outbreak, Canard had focused on derstand the coronavirus RdRp better, they the structure–activity relationships of nucle- found that the polymerase by itself could not otide analogs used to treat HIV and other explain the disconnect. Virologist François viruses. Like many virologists, he was inspired Ferron, a staff scientist at CNRS, has worked by the near miss with SARS to start new on coronavirus replication since the SARS research programs. outbreak. “The viral RNA polymerase is quite

38 ASBMB TODAY MAY 2020 FEATURE

Following up on that work, the French group zeroed in on how nsp14 worked in a test tube. “We didn’t work on the virus, like Mark Denison was beautifully doing,” said Canard. “We just concentrated on that enzyme … and we found out that it could actually excise ribavirin.” The CNRS team published an enzymology study that confirmed that nsp14 can identify and remove mismatches between bases at the end of a growing copy of viral RNA; in 2018 they followed up with confirmation that when ribavirin is incorporated in a growing RNA strand, nsp14 protein can scoop it out of the Virologist François Ferron, a staff scientist at CNRS, stalled strand, letting replication resume. has worked on coronavirus replication since the SARS As the researchers worked out the enzy- outbreak. mology, a pressing question arose: Did their findings mean that all nucleotide analogs would be useless against coronaviruses? loose, meaning it has a tendency to make a lot “This question is actually key right now in of mistakes,” he said. “Maybe a little bit more the development of nucleoside analog inhibi- than the regular (cellular) RNA polymerase.” tors,” Canard said. This led researchers to suspect that coro- naviruses might have some way of recognizing Evading viral proofreading and correcting errors. This is where remdesivir comes in. An international team conducting a study “From my perspective, the (remdesivir) of the SARS viral genome identified a number story started in 2013,” Denison said. “We of potential RNA-processing enzymes based had discovered that coronaviruses encode the on their homology to other known enzymes. only known RNA proofreading system … In 2006, Canard’s group worked with mem- (and) I wanted to test whether there were any bers of that international bioinformatics team nucleosides out there that could be active in to show that one of those enzymes, like its the setting of coronavirus proofreading.” homologs, could cleave double-stranded Denison heard from Cameron about a RNA and was required for successful viral research collaboration with Gilead Sciences. replication. Cameron was working on understanding the “There was a speculation that (coronavi- precise mechanism of action of a group of ruses) might encode a proofreading function nucleotide analogs the company was using to that would allow them to stabilize a big treat hepatitis C, an RNA virus that infected genome, and there was a predicted place in tens of thousands of people each year. the genome where that might occur,” Denison Before the race to drug hepatitis C, explained. “That really led us to try the genet- according to Adrian Ray, a medicinal chemist ic experiments.” who used to work at Gilead, “The nuc space In 2007, his group found that in viruses for RNA polymerases and for RNA viruses lacking the protein encoded at that same was really not heavily explored.” location, a protein called nsp14, coronaviruses In the course of trying to beat competitors from a mouse model virus accumulated mu- to the lucrative hepatitis C market, Gilead had tations at a rate similar to other RNA viruses. developed a large library of RNA-dependent And strains of the virus without nsp14, they RNA polymerase inhibitors, including the found, were sensitive to ribavirin. molecule that would come to be known as

MAY 2020 ASBMB TODAY 39 FEATURE

The candidate molecules arrived. Den- ison and his trainees had no idea what they were. But they went ahead and tested them. What they found was exciting: In mouse cell GILEAD SCIENCES culture, the drugs could block coronavirus replication. “So we asked our Gilead collaborators, ‘What is that (compound)?’ They said, ‘We’re not going to tell you, but we’re going to send you 60 prodrugs, chemical modifications of the same compound,” Denison said. One of that second batch of molecules proved to be remdesivir. Graduate student Maria Agostini, who had recently joined Denison’s lab, was one of the researchers who Maria Agostini, a grad student in Mark Denison’s lab, was worked on understanding its strong activity. one of the researchers who worked on remdesivir. “We’ve worked with a couple of potent compounds, but remdesivir was really one of remdesivir. A compound closely related to the first I had worked with,” Agostini said. remdesivir had made it to early clinical tests “When you were looking at the cells, you for hepatitis C, but had faltered, in part be- could visually see less evidence of viral repli- cause like remdesivir it needed to be adminis- cation going on.” tered by injection. Gilead changed strategies, Whereas cells in her control dishes were buying a biotech startup to gain access to the visibly infected, suffering bad cytopathic startup’s orally available nucleotide analog, effects, the cells treated with remdesivir after which became a key component of Gilead’s infection survived well. By growing many hepatitis C cocktail. generations of the virus in cells treated with Working with the RNA-dependent RNA subtherapeutic concentrations of remdesivir, polymerase from , Cameron had selecting for mutations that would let the begun work that would show that the mol- virus evade the drug, Agostini and colleagues ecule was a non-obligate chain terminator, a Erica Andres and Clint Smith demonstrated type of nucleotide analog that the polymerase that it would take mutations in the viral poly- ought to be able to incorporate into a grow- ing strand and keep going — but could not. Intrigued, Denison contacted Gilead to ask permission to try that approved drug, called sofosbuvir, against mouse coronaviruses. “That was their world-changing drug that cured hepatitis C,” Denison said. “They weren’t going to let a little-known virologist working on a coronavirus use their drugs.” But after a series of introductions by Cam- eron and several discussions, Gilead agreed to let his lab work with a different series of molecules, the ones that had been developed in-house and shelved for hepatitis C. They had shown promising results in early studies as a candidate treatment for other viral infec- Mathias Götte’s lab in Canada has studied how remdesivir tions, including against Ebola virus. works on polymerase enzymes from the MERS coronavirus.

40 ASBMB TODAY MAY 2020 FEATURE GILEAD SCIENCES

In a remdesivir production facility, Gilead employees work in personal protective equipment.

merase to confer resistance to remdesivir — and that those mutant viruses were less able to infect hosts than the wild type. Gilead supplied the drug free of charge, and the National Institutes of Health funded the researchers through a program aimed at developing treatments for emerging infectious diseases. “This points out the value of collaborative science,” Denison said. “This was a company that committed to helping us do this when no one was interested in coronaviruses, and a grant mechanism that allows some flexibility in terms of expanding it.” Recently, Mathias Götte’s enzymology lab in Canada has looked at how remdesivir works Andrea Pruijssers directs the antivirals research program in on polymerase enzymes from the coronavirus Mark Denison’s group. that causes MERS. Researchers in Götte’s group determined that the compound stops the enzyme keep going for a few more cycles the polymerase, acting as a chain terminator but then causes it to stall. Researchers suspect — but not immediately. that the molecular stumble may be caused Andrea Pruijssers, a virologist who directs by an unusual structure in the template-copy the antivirals research program in Denison’s RNA duplex. group, said, “(Remdesivir) somehow evades “They think that, at that point, the nucleo- recognition by the proofreading enzyme.” side analog that has already been incorporated Instead of stopping the polymerase as soon is shielded from the proofreading enzyme,” as it is incorporated, remdesivir seems to let Pruijssers said.

MAY 2020 ASBMB TODAY 41 FEATURE COURTESY OF ANDREA PRUIJSSERS

Members of the Denison lab at a less pandemic time: (Left to right) Maria Agostini, Jim Chappell, Jennifer Gribble, Andrea Pruijssers, Laura Stevens, Tia Hughes, Mark Denison, Xiaotao Lu.

In the lab of Denison’s longtime collabo- that causes COVID-19. rator Ralph Baric at the University of North “It’s interesting,” Pruijssers said. “(NHC) Carolina, Chapel Hill, researchers found doesn’t act as a chain-terminator. It incorpo- that remdesivir was an effective treatment rates into the genome and then causes lethal for mice infected with SARS. Those results mutagenesis.” were promising enough to advance remdesivir Through its ability to base-pair with more into a study of MERS in monkeys, published than one nucleotide in the complementary in February by researchers at the Rocky strand, NHC introduces a cascade of errors in Mountain Laboratory of the NIH. The drug successive rounds of replication. Eventually, showed some benefit in reducing the severity viral progeny don’t have the information they of the illness, provided the monkeys were need to make a new virus. treated prophylactically. Whereas remdesivir stops the polymerase In terms of drug development, remdesivir in its tracks, causing few new mutations, deep “has sort of met every milestone along the sequencing experiments in the few viruses way, from our perspective,” Denison said. that emerged after NHC treatment showed that the drug causes numerous mutations. Similar drug class, different result “We identified two compounds that struc- When Pruijssers started in the Denison turally fall into the same class of nucleoside lab in 2017, they had another molecule to analogs but act very differently on coronavi- investigate, beta-D-N4-hydroxycytidine, or ruses,” Agostini said. NHC for short, that was being tested at the They were particularly encouraged because Emory Institute for Drug Development as a the work showed that NHC has some thera- potential broad-spectrum . peutic efficacy in the mouse model of MERS Agostini led the first study of that drug — and that it can block even remdesivir-resis- as well, showing its activity in tissue culture. tant strains of coronavirus. In March, researchers from the Baric and In an interview in early March, Pruijssers Denison labs published a followup in Science said NHC was still relatively untested as a thera- Translational Medicine, showing that NHC peutic. “It’s hard to develop a mutagen, because can block replication in the viruses that cause the FDA doesn’t usually like the idea of muta- MERS and SARS — and also the coronavirus tion unless it’s for a life-threatening disease.”

42 ASBMB TODAY MAY 2020 FEATURE

But the pandemic changed things. Emory in April as a trial at the China–Japan Friend- has partnered with Miami biotechnology com- ship Hospital in Wuhan concluded. (Other pany Ridgeback Biotherapeutics to develop the trials, sponsored by Gilead, the NIH and the molecule, and Emory has filed an application World Health Organization, launched later.) with the U.S. Food and Drug Administration First, animal data from other coronavi- for permission to begin first-in-human trials to ruses suggest that the drug is most effective test the molecule’s safety in humans. when it’s delivered prophylactically, after the Denison described the pivotal role Agos- animals are exposed but before they begin to tini played: “In five years of graduate school, develop symptoms. In the context of a global Maria tested and developed the detailed in vitro pandemic, this would be difficult to achieve analysis on two potential drugs to treat this pan- in humans. At some point in the course of in- demic coronavirus — and got them all the way fection, an extremely strong immune response through that in vitro preclinical development.” begins to do more harm than the virus; at that It’s a remarkable and highly unusual feat. point, Denison said, it may be too late for an Drug development is known for its high antiviral to help. failure rate. Of course, many drug candidates Second, the data from the first few trials that appear promising in preclinical studies are likely to be nuanced and require careful falter in large human trials. interpretation — which Denison worries public discourse is not well prepared for. Early Facing the pandemic human trials of remdesivir’s sister compound After arguing for a decade that the world in hepatitis patients showed dramatic differ- must be ready for a pandemic, Denison said ences among individuals in antiviral response, in early March, it was strange to be facing it. and remdesivir itself showed limited benefit “It’s really weird that we worked on this compared to other candidate therapies in a for the past six years, and the drugs were just clinical trial in Ebola patients. “People tend getting through, and they were just ready to to have a winner or loser mentality,” he said. go,” he said. “We’ll see what the outcome is.” “They’ve called remdesivir ‘that failed Ebola He has some concerns about how to inter- drug,’ right? It didn’t fail in the Ebola trial; it pret data from clinical trials testing how well just wasn’t advanced because it didn’t show as remdesivir works for COVID-19 in humans, much benefit as the other two compounds.” the first of which were expected to be released Whatever the outcome of clinical trials of remdesivir and NHC, Denison said he hopes this crisis will underline the importance of funding research into potentially pandemic viruses before outbreaks begin. “Trying to maintain basic investigations and drug development against something

COURTESY OF ANDREA PRUIJSSERS that’s a high, high, high impact but low, low, low probability is really hard in our world. Re- ally hard,” he said. “We’ve just never given up on this idea that we had to have these things ready and have them in the bucket.”

Laurel Oldach (loldach@ asbmb.org) is a science writer for the ASBMB. Follow her on Twitter @LaurelOld.

Denison, pictured in January with the spring 2004 issue of the Vanderbilt University Medical Center’s Lens magazine.

MAY 2020 ASBMB TODAY 43 FEATURE

Scientist uses community organizing skills to mobilize researchers against COVID-19

By Laurel Oldach

round the world, scientific difficult task of putting the list into thousands of volunteers registered researchers are seeking ways to the hands of people who need it. and the tide of new arrivals slowing, Achannel their abilities and ener- This isn’t Wells’ first experience Wells and colleagues he met during gy in the fight to contain the spread responding to a disease outbreak. In a stint as a Society for Neuroscience of COVID-19. When Michael Wells, 2015, as a postdoc starting out in policy fellow are working to make furloughed from his laboratory and a lab that studied brain organoids, local governments aware of the wanting to help, launched a nation- he had planned to work on autism. resource. al volunteer database, it took off But then the Zika virus outbreak quickly. spreading from Brazil to other parts Local responses “I think it’s grown about 100 of North and South America began With vacillating federal gov- people since we started talking,” to make headlines. With much of his ernment leadership, much of the Wells, a postdoctoral fellow at the extended family living in El Salvador, country’s response to the COVID-19 Broad Institute in Boston, said 15 Wells said he saw “an opportunity epidemic is happening at the state minutes into a phone interview on for me to not only learn all these new and local level. Wells sees that March 20, just two days after the techniques, but also potentially do reflected in how he has distributed online questionnaire for the database something that could have a direct the volunteer database to those who was launched. impact on people in my life.” might be able to use it. After four years of postdoctoral “It seems like individual counties Building a database research and several publications in individual states are responding Twelve years ago, Wells ran the on Zika and the developing brain, differently to the virus,” he said. Obama primary campaign on his Wells recently began to consider For some, that is a source of college campus at the University of seeking a faculty job. But once again, some comfort. Derek Crowe, a Notre Dame and, during the sum- a virus changed his plans. Scrolling graduate student at the University mer, he worked for Obama in Ohio. social media feeds for news about of Rochester is working to coordi- Using skills honed during his time as the pandemic after his laboratory nate local volunteers with the health a campaign volunteer and an Obama shut down, Wells saw calls from department. fellow, Wells and an ad hoc team of diagnostic labs in other parts of the “I actually trust the local govern- collaborators have recruited more country for people with molecular ment a lot more than the national than 8,300 researchers who want to biology skills. He waited a while government to get things done,” contribute to the COVID-19 effort. for something similar to pop up in he said. Seeing local government, The list includes volunteers’ lo- Boston but then decided to build it healthcare workers and hospital staff cations, laboratory experience, access himself and, while he was at it, to working around the clock “indicates to supplies that could be donated make a tool that could be used across to me that there is leadership that and — by request, for vetted public the country. He recruited musi- approaching this situation with seri- health organizations only — names cian friends to promote it on social ousness and professionalism.” and contact information. Wells and media. With the situation changing rap- his team are now tackling the more Word spread quickly. Now, with idly, it can be difficult for members

44 ASBMB TODAY MAY 2020 FEATURE COURTESY OF MICHAEL WELLS

“If the issue Michael Wells is using the community organizing skills he learned in an election campaign to build a database of isn’t that there researchers who want to help in the fight against COVID-19. aren’t enough of the public to find current infor- ry in Island County, Washington, in mation on testing. Crowe said that search of laboratory personnel and a tests but that doctors in his area are only testing hospital administrator in Laurel, Mis- people with symptoms, because of a sissippi, who needed help sourcing there aren’t shortage of diagnostic supplies. UV lamps have requested access to “If the issue isn’t that there aren’t regional subsets of the database. That enough human enough tests but that there aren’t diversity of purpose, Wells said, is enough human resources to get them part of the point. resources to get done, we want to be able to fill that “We don’t know what they are gap,” he said. going to need it for; they don’t know them done, we Monroe County, New York, what they are going to need it for where Rochester is located, has two yet. The focus is on getting it into want to be able large universities, and most of the their hands and letting them figure volunteers in the area are graduate out the best way of activating all this students or principal investigators scientific energy.” to fill that gap. at the University of Rochester or ” Rochester Institute of Technology. According to Wells, who lives in Laurel Oldach (loldach@ Boston, the larger concern is for less asbmb.org) is a science writer for the ASBMB. Follow densely populated areas with a lower her on Twitter @LaurelOld. concentration of universities and biotechnology companies. Already, a public health laborato-

MAY 2020 ASBMB TODAY 45 FEATURE

Researchers retool genomics labs to provide virus testing By Laurel Oldach

esearchers at San Francisco’s In- novative Genomics Institute an- Rnounced on March 30 that they were opening a COVID-19 testing center able to provide at least 1,200 tests each day for patients in San Francisco. The IGI is one of several academic institutes whose laborato- ries have switched focus to contribute to the COVID-19 response. Labs that conduct high-through- put genome sequencing at IGI, Postdoctoral fellows Enrique Shao and Jenny Hamilton work in a converted diagnostics lab, now Boston’s Broad Institute and San being used for COVID-19 testing, at the Innovative Genomics Institute in San Francisco. Francisco’s Chan-Zuckerberg Biohub retooled recently to provide clini- IGI, put out a call for volunteers and nobody’s doing high-throughput cal testing for SARS-nCoV-2, the in mid-March after hearing about sequencing experiments right now, virus that causes COVID-19. These the diagnostic testing effort at the we’ve been able to quickly reconfig- overhauls by volunteer scientists and neighboring Biohub. Within hours, ure the room.” administrators added capacity for 800 members of San Francisco’s bio- Using liquid-handling robots and thousands of tests and rapid turn- medical research community, mainly reagents bought and donated from around time to help public health graduate students and postdocs, multiple sources, Hamilton and her departments determine how many responded. team set up and validated a mostly people are infected. Jenny Hamilton, a postdoctoral automated pipeline for extracting Jennifer Doudna, a professor at fellow in Doudna’s UC Berkeley RNA from patient samples and the University of California Berke- lab, led the technical team that conducting a standard reverse tran- ley and a Howard Hughes Medical transformed the genomics core into scription polymerase chain reaction, Institute investigator, founded and a clinical testing laboratory, and or RT-PCR, assay for SARS-nCoV-2. leads the IGI. validated the test results, in just two Advisers from industry helped the “Imagine setting that up — a weeks. researchers program the robotic process that would normally take “The room that we’re using pipeline. months to years — in a couple was previously part of a genom- Some widely used reagents, such weeks. It’s really extraordinary,” ics core facility on campus, and as disinfectant and qPCR master Doudna said in an IGI press release. because of that it had some robotics mix, have been difficult to come Doudna and colleague Fyodor for high-throughput experiments by. According to Alexandra Amen, Urnov, also a UC Berkeley profes- already installed,’ Hamilton said. another postdoc in the Doudna lab sor and a technical director at the “Because the campus is shut down who helped run the lab-conversion

46 ASBMB TODAY MAY 2020 FEATURE

effort, “The hardest thing is sourcing PR systems from microbes, improve Health have relaxed some restrictions materials that are in low supply.” genome editing tools, genetically on who may test for and report new As testing began, volunteer engineer crops and diagnose human cases of the novel coronavirus. researchers planned to operate the di- diseases. Nonetheless, some logistical barri- agnostic lab, testing samples collected “All of our laboratories do PCR ers remained in place as ASBMB Today from patients at UC Berkeley’s med- every day,” Doudna stated in the IGI went to print in mid-April. The journal ical center and other hospitals in San release. “But for this test we need to Nature reported on April 9 that even as Francisco’s East Bay. They hoped to go above and beyond to ensure we California testing pipelines remained

MAX & JULES PHOTOGRAPHY / INNOVATIVE GENOMICS INSTI / INNOVATIVE MAX & JULES PHOTOGRAPHY return results of 1,200 to 4,000 tests can provide accurate detection.” backlogged, many hospitals had daily within 24 hours, improving on Researchers didn’t know initially declined free diagnostic testing from turnaround times that Kaiser Health whether the effort was entirely legal. the IGI because the lab lacked software News reports reported could take Clinical laboratories must be licensed that commercial diagnostic labs use for three days to a week. As for funding, by state and federal governments to electronic health records. the team said through a spokesperson, ensure that they report valid results.

“We hope to fund the IGI’s testing But given the COVID-19 public Laurel Oldach (loldach@ lab through donations to enable us to health emergency, the IGI release asbmb.org) is a science writer for the ASBMB. Follow serve those without insurance.” stated, the Centers for Disease her on Twitter @LaurelOld. Ordinarily, the institute focuses Control and Prevention, the U.S. on genome engineering. Its research Food and Drug Administration and programs work to isolate new CRIS- the California Department of Public

COVID-19 resources and news from the ASBMB

asbmb.org/career-resources/online-resources

MAY 2020 ASBMB TODAY 47 FEATURE

We are doers. We want to get involved. How“ basic scientists at University of Washington contributed ” to diagnostic testing for COVID-19

By Laurel Oldach

he basic science laboratories at by government-certified laboratory fied technicians. But the roughly the University of Washington technicians. However, testing rates 150 volunteers — most of whom T School of Medicine in Seat- for the novel coronavirus SARS- are graduate students, postdoctoral tle are under the same roof as the nCoV-2 lagged badly in the United fellows or junior faculty –helped clinical care facilities but separated States compared with other coun- with ancillary tasks, from restocking by long hallways. In early March, tries. As the number of people with shelves to logging samples as they when Seattle was the first American respiratory symptoms grew, demand enter the lab. hotspot but before Washington’s for testing strained the university’s “Something that’s very heart- governor issued shelter-in-place laboratory medicine department. warming is everybody pulling orders, those halls were very quiet. The first shortage was hardware; together within the University of Faculty members and trainees with the interim chair of the pathology Washington to make this happen,” children and those at high risk of department circulated a request said Paul Scott, the chair of the uni- contracting COVID-19 shifted to for specific PCR machines early in versity’s pharmacology department. remote work, and the remaining March. “It started with an idea, and it’s scientists carefully avoided close In subsequent weeks, the lab’s expanded very quickly into action.” contact. personnel became overwhelmed. As Meanwhile, other members of “It’s a really weird combination the university prepared to launch the University of Washington com- of intense emotional distress and a drive-up testing center, Gordon munity found ways to contribute to being deserted. Normally, I think, said, “there was the recognition that the outbreak response. Rapidly-de- when a place is deserted, it’s empty the state testing lab, in combination veloped message boards within the of emotion, too,” said Sharona Gor- with the university’s testing lab, just UW community matched healthcare don, the university’s associate dean didn’t have the capacity — and that workers in need of childcare and for research and graduate education. outside-the-box thinking was going people at higher risk of infection At a time when normality felt to be necessary.” with delivery needs to people who like a distant memory, Gordon Luckily, the UW community want to help. Research labs crowd- became one of many basic scientists includes a large number of scien- sourced reagents and coordinated helping with an unusual effort: tists who know their way around a donations. The stringent Food and recruiting volunteer scientists to help thermocycler. After consulting with Drug Administration protocols that keep diagnostic labs running during faculty and department chairs, the clinical laboratories follow often Seattle’s COVID-19 outbreak. dean of research sent an email on specify the manufacturer and type of March 12 requesting volunteers reagents or consumables that may be Reassigning basic scientists under age 40 to help out in the labs used. One Friday, the UW virolo- Normally, basic scientists and at the department of laboratory gy department put out an urgent clinical lab workers overlap very medicine. request for pipette tips; that Saturday little. Because of federal regulations, The diagnostic tests themselves morning, it reported that donations diagnostic tests must be carried out still had to be performed by certi- from academic groups, including

48 ASBMB TODAY MAY 2020 FEATURE BRANDON HOLMES / UNSPLASH

an influenza surveillance study, had contact was difficult for the scientists allowed it to continue diagnostic tests she knows to follow, even though its Laurel Oldach (loldach@ asbmb.org) is a science until a manufacturer’s shipment importance was clear. “We are doers. writer for the ASBMB. Follow came in. We want to get involved,” Gordon her on Twitter @LaurelOld. Before Washington’s governor said. “But if you put 100 people in a issued a shelter-in-place order on room to get involved, you would be March 23, Gordon said, the advice spreading the virus.” to stay at home and avoid social

MAY 2020 ASBMB TODAY 49 PERSPECTIVES

On the front line: Pandemic insight from a health care worker By Justin R. Lovett

orking in a hospital emergency When I am caring directly for a As the number of patients department in the midst of patient who shows symptoms and increases, emergency departments Wa pandemic, I am learning a meets criteria for COVID-19 testing, and critical care units have become new routine. As COVID-19 num- I have to wear a gown, gloves, an overcrowded, and we have extra staff bers increase in Texas, our protocols N95 respirator, boot covers, shoe on standby. In the month of March, get stricter. The state had more than covers over the boot covers, goggles, I worked a total of eight shifts. In 7,300 confirmed cases on April a hood, a face shield and a surgical April, I am working 16 shifts to help 6, and we have seen dozens at the mask over the respirator. This is a with this influx of patients. hospital where I work. Our infectious lot to put on each time you enter a I sometimes feel it is a matter of disease team is working diligently to patient’s room. “When will I contract this virus?” as ensure the safety of the staff, so our Most health care workers share a opposed to “I hope I don’t contract protocols are changing continuously. concern about what happens when this virus.” That’s an ominous feeling. Before the pandemic, I got we run out of PPE: When we can The community has been incredibly dressed at home and then drove to no longer wear PPE with infectious supportive of the ED staff, sending the hospital for my shifts as an emer- patients, what do we do? Already, we us food and other tokens of apprecia- gency medicine technician. Now, I get one single-use surgical mask for tion. Though the feeling is ominous, have to wear civilian clothes to the a 12-hour shift. That mask provides it is reassuring knowing we have the hospital and then change into scrubs standard droplet protection. How- support of our community. and put on shoe covers, a surgical ever, when we have COVID-19 pa- In addition to working on the mask and a surgical cap. At the end tients at the hospital, we need to take front line in the ED, I am a student, of the shift, I change back into my precautions against both droplets and and the pandemic also has affected civilian clothes and go home, strip airborne particles. N95 respirators how my classmates and I learn. This those clothes off at the laundry, show- are the most secure protection against semester, I am enrolled in 18 hours er and put on fresh clothes. Then I infectious particles. The hospital of coursework ranging from Bio- am free to walk about my house. I allots a single N95 mask for 8 hours chemistry II to Calculus. It has been do this to contain any potentially of use. We pick and choose when we a struggle for students and professors infectious particles I may have come wear them, because we cannot order to transition these intensive courses into contact with. any more. to a distanced platform and ensure The personal protective equip- We have a strict no-visitor policy the same quality of instruction. The ment described above is standard for in the ED; all patients and staff are professors have been amazing and interacting with any patient, whether screened prior to entry, even patients continuously check in on us along or not they show COVID-19 symp- coming in on an ambulance. The the way. Now we have lecture videos toms. If you enter the emergency screening includes a temperature and Zoom office hours. My main department with a toothache, you’ll check and a questionnaire; the results concern beyond adjusting to an see us wearing scrubs, shoe covers, determine which patients need to online platform is my senior research. caps and masks. This is for safety so wear a surgical mask. Before we had Research is an integral part of our we do not spread particles from our the screening process, patients would curriculum in the chemistry depart- previous patients to new ones. PPE, grab handfuls of masks and leave ment. Before spring break, I was in always important, is essential during with them. Each employee gets a the purification process of important this crisis; however, faced with short- dated sticker that we must display samples for my senior thesis, which ages everywhere, we are rationing our stating we have been screened on the had to stop. Luckily, I am working PPE gear. current day. with a sturdy protein, so my fingers

50 ASBMB TODAY MAY 2020 PERSPECTIVES JUSTIN LOVETT

Justin Lovett, at work as an emergency medicine technician in a Texas hospital, wears a scrub cap his grandmother made for him.

are crossed that I’ll return to viable ily table for Mom and Grandma’s and we make sacrifices. samples. Otherwise, it’s back to Southern home cooking, to laugh, This new reality has taken an square one of purification. play games and hug one another like emotional and physical toll on all Working increased shifts in the we used to do. We all communicate health care workers. However, we are ED means more time away from daily via group message and the banded together as a community to loved ones. We have to keep our occasional phone call. This time has help fight this pandemic. distance because we work in a high- been especially hard on my parents, risk environment and do not want to who have four children working in Justin R. Lovett is a infect anyone. We don’t know how health care. We’ve planned a group senior chemistry major at long this will last. I wonder some- Facetime so we can see each other. Stephen F. Austin State University and serves times when normalcy will return. My grandmother is keeping busy by as the recruitment chair Because of my work, I made the making us all scrub caps. for SFASU’s ASBMB cautious decision to remain away I am thankful for my team in the Student Chapter. Justin works in Odutayo from my family. I have not seen them emergency department. We took an Odunuga’s lab, where he in person for a couple of weeks, and oath to put patients first and to help enjoys researching the biophysical properties I wonder when I will get to see my others. That is what we are doing, of protein structure and siblings, nieces, nephews and parents. working diligently to provide the best function. He plans to pursue an academic I look forward to the day when care we can to all patients. We take career in biophysical we can all gather around the fam- risks caring for COVID-19 patients, chemistry.

MAY 2020 ASBMB TODAY 51 PERSPECTIVES

Quarantined thoughts

By Makayla K. Portley & Marya S. Sabir

e used to think science stopped for no one and noth- Wing, but now that feels only partially true. During these uncertain times of mandated social distancing and a 24-hour COVID-19 news PORTLEY COURTESY OF MAKAYLA cycle, we’ve been reflecting on how different our lives are compared to four weeks ago. Back then, we were in the lab working diligently on finishing con- firmatory PCRs for a large whole-ge- nome sequencing study and organiz- ing patient samples for another large study. Now, our days are filled with daily 9 a.m. lab meetings via Zoom, other conference calls (so many vir- tual platforms — and let’s be honest now, we all have a favorite), learning bioinformatics, and making loaf after With her lab closed for now, Makayla Portley can spend more time in her kitchen baking tasty treats. loaf of fresh sourdough bread (yum). As we do our best to navigate • We miss the sight of the qPCR Just kidding, no one ever smells these unprecedented times, we machine generating its multicol- Trizol because it is always used remember the intimate idiosyncrasies ored curves in real time. Even in a perfectly ventilated fume of being a bench researcher and how when the experiment fails, it’s hood through which no odor much these are tied to our identities still pretty. escapes … And we don’t miss it. as scientists. Each researcher has a • We miss the way that our tube • We miss the smell of the baked unique set of senses with which we racks look when they’re finally goods shared in the lunchroom perceive the fascinating world of organized and all oriented the by our wonderful lab mates. bench science. Here are some sensory same. exact. way. It takes dedica- Another chance to smell baked wet lab experiences that we’re missing tion, but it is fulfilling. bread. during our mandatory telework days. • We miss the sight of the lab’s • We miss the odor of rotten eggs messy benches after a hard day’s and burnt rubber — oh wait, Sight work. that’s just beta-mercaptoethanol • We miss the fluorescent lighting being used outside the hood. of the lab, even though it usually Smell causes us headaches after about • We miss the aroma the autoclave Touch four hours. produces during its cycle, which • We miss the cool benchtop on a • We miss seeing other passionate is mysteriously similar to bread warm day. scientists (AKA fellow nerds). baking. • We miss the weight of a pipet in Our political science major • Quite inexplicably, we miss the our hands. friends don’t share the same zest faint scent of mice coming from • We miss the weird feeling of hand for neurological disease research the behavioral testing room. sweat inside our non-latex gloves wormholes. • We miss the smell of Trizol. (not that much, though).

52 ASBMB TODAY MAY 2020 PERSPECTIVES

• We miss our itchy, too-warm lab excited about successful exper- Beyond that, we’re hoping this coats. iments: gasps or general praise one doesn’t apply to anyone these • We miss holding hundreds of of science, cells, an antibody — days since mouth-pipetting is strong- tubes as we attach adhesive labels whatever is involved. ly frowned upon. to the lids. We suspect this fond- • We miss hearing the freezer Here’s to being able to do bench ness will be short-lived when we beeping madly to alert us that it’s science again — soon. return to the lab. gotten slightly too warm. Coin- cidentally, that sound also haunts Hearing our dreams. Makayla K. Portley (makaylaportley@ • We miss the sounds of our Max- • We miss the only sound in the gmail.com) is a well extracting DNA from brain lab at 2 a.m. — the hissing of postbaccalaureate fellow studying neurogenetics. tissue (it’s just normal machine liquid nitrogen tanks in the Follow her on Twitter @ sounds for which the exact ono- corridor. We only recently makaylaportley. matopoeia is not yet coined). realized this wasn’t coming from • We miss the sound of the cell lab ghosts. counter focusing. We don’t know Marya S. Sabir (msabir@ why, but it’s such a specific, whir- Taste asu.edu) is a graduate student studying ring sound that holds a special We miss the taste of the gummy neurogenetics. Follow her place in our hearts. bears that come with our primer on Twitter @marya_sabir. • We miss the sounds of scientists orders from Eurofins.

We miss holding hundreds“ of tubes as COURTESY OF MARYA SABIR COURTESY OF MARYA we attach adhesive labels to the lids. We suspect this fondness will be short-lived when we return to the lab.”

Marya Sabir works in the lab before the COVID-19 pandemic shuttered her institution.

MAY 2020 ASBMB TODAY 53 PERSPECTIVES

A new city, a new job and a global pandemic Plus, a natural disaster for good measure By Elizabeth Stivison

n the evening of Feb. 27, my stubborn dog lets Milo lick his face. tery, and he came running when he friend Germaine and I finished She had just arrived home from a saw me. Opacking up my U-Haul. We trip and was jetlagged, but she went I texted the group and everyone hugged my roommates goodbye, to look too. I texted Kelly, owner of was overjoyed and relieved. I felt and, along with my dog, Milo, piled a big curly-furred puppy. She was like I was George Bailey in the end into the truck to leave New York out of town, but her husband and of “It’s a Wonderful Life,” when his City. daughter went to look. I called my friends and family come together to I’d lived for seven years in Upper roommates, my friend Joy (by then save him. My pup, the little goofy Manhattan while earning my Ph.D. in tears) and my neighbor Amanda. love of my life, had been saved by at Columbia University and nearly They all looked. My labmate and my community. my whole life within a few hours’ friend Amr drove down from the But, unlike George Bailey, I bus ride of the city. As much as I Bronx to look in his car. didn’t earn this support in any real complained about NYC, it was my Suddenly, it seemed like every way: I never saved any bank from the home. I felt excited, but also a bit dog owner and friend in my neigh- Depression or helped anyone buy nervous, to be leaving to start my borhood was looking. Numbers I their home. postdoc at Vanderbilt University. didn’t even recognize were texting me And, perhaps stupidly, I was As we drove away, I was filled where they were looking. A group leaving this caring community in a with love for my community and text emerged. As I sat in my hotel, few weeks for a new job. neighborhood that I was leaving. waiting until it was time to leave for What I didn’t know was that my the airport, people were updating Welcome to Nashville new city and new postdoc life would each other on who had seen Milo Germaine and I arrived at my be nothing like I expected. Within and where. new apartment in Nashville on a few weeks’ time, I would narrowly “He’s in the northwest corner of March 2. We piled all my boxes escape a devastating tornado, and the playground, but he ran off.” in the living room, set up my bed the COVID-19 pandemic would “Saw him running north from and an inflatable mattress for her, turn the world upside-down. the subway stop.” returned the U-Haul and went to “Oh, Milo! That little white dog? sleep. Three weeks earlier I know that dog! The girl loves him During the night, I woke to the “Milo got out of his leash and so much,” a guy at the corner bodega sound of sirens. “Man, I was really ran away!” my dog sitter, breath- told my friend. hoping Nashville would be quieter less from running, told me on the Roger, the owner of a golden re- than New York City,” I thought. phone. I was down in Nashville triever who Milo loves, looked until In the morning, everyone was looking for an apartment. almost 2 a.m., even though he was texting me: my mom, my old room- Panicked, I hung up, bought flying to Mexico a few hours later. mates, my friends. “I heard there the first plane ticket home, and Around 5 a.m. the next day, as was a tornado in Nashville. Are you immediately started calling people I was in the airport waiting for my OK?” I told them all I was fine. I knew from the dog park. I called flight, the texts and updates started I went into the living room and Katie, whose small Muppet-like dog again. By the time I landed, I was said, “Germaine, I think there was likes to wrestle with Milo; she went armed with maps of Milo sightings. a tornado last night … nearby.” We to look. I called Trena, whose big I finally spotted him in a ceme- looked on the map together and saw

54 ASBMB TODAY MAY 2020 PERSPECTIVES COURTESY OF ELIZABETH STIVISON

Elizabeth Stivison poses with her dog, Milo, in front of the U-Haul she used for her move to Nashville. that it was about 3 miles from us. “I energy out and so I could meet other our dogs at the same time. didn’t know that’s what those sirens dog owners. I chose a building with meant!” I said. common rooms, a café, an office with Things changed quickly Welcome to Nashville! Internet and a coffee machine, next On March 13, my former lab- The next day, after saying good- door to a big dog park. mates forwarded me an email that bye to Germaine, I heard on the I was excited to meet people and said: “The hospital has a critical need radio about a case of COVID-19 in see what the Music City was like. I for surgical masks … for all labs that New York, and it was near where I decided I’d help after the tornado by have surgical masks, please bring used to live and work. I texted my supporting local businesses by my them down … there will be a box former roommate, Katherine. patronage. I went to an art event and for depositing them when you enter “I know! It’s starting …,” she met a sweet and kind couple whose the suite. PLEASE BRING THEM wrote. street was destroyed by the torna- NOW.” “Wash your hands!” I told her. do, but somehow their house was The next day, they texted me that undamaged. We exchanged numbers the lab at Columbia was closing. Amenities for making acquaintances and planned to meet up soon. I emailed my new boss at Vander- My apartment in New York had I went to the dog park each day bilt to make sure I could still come in been a four-bedroom, so I knew I’d and slowly began meeting people. on Monday for my first day of work. want to be able to see people in my I loved the feeling of arriving and He said I could but that the labs new building in Nashville. I also finally recognizing faces, dog and might close soon, so I should start knew I’d need a dog park nearby human. I exchanged numbers with a thinking about remote projects and so that Milo could play and get his few people and made plans to bring only do minimal lab work that was

MAY 2020 ASBMB TODAY 55 PERSPECTIVES

essential to my project. Our PI told us that, so far, “Is this the correct media?” I’d March 16: My first day of work Vanderbilt considers labs essential. ask, holding the bottle up to the in my new lab! Also the first day my As a lab, we decided a few things. camera. old lab at Columbia was officially First, that we will try to not overlap One of the few times my boss shut down. in lab. Second, that we will probably and I were in lab at the same time “I think the lab will be closed be shut down soon, though we don’t (we both wore masks), he was teach- soon,” my boss told me. know when. Third, that we’ll have ing me how to freeze cells for liquid I started making cell lines to use Zoom lab meetings once a week to nitrogen storage, and while waiting later, and Vanderbilt banned Ph.D. keep us all connected. “So we don’t for the media to be ready, he told me students from coming in. come back in a few months a bunch about his daughter. My apartment building closed of strangers!” our PI said. “She’s been bugging us to get a all the common rooms, and the city After learning how to do tissue pet. She wants a dog so badly. But closed the dog park. culture, I started going into lab every we agreed she could start with a On March 22, the mayor of few days, always checking to make mouse,” he said. Nashville issued a safer-at-home sure no one else would be there, and “You should get her a hamster order. I texted another postdoc in my did my work alone. When I couldn’t instead!” I said with conviction. lab, “What does this mean for lab?” figure something out, I’d Zoom with “Mice are stinky. Hamsters don’t She didn’t know. my boss and show him my problems. stink as much. I always had hamsters COURTESY OF ELIZABETH STIVISON

Elizabeth Stivison, at left, her friend Germaine and Milo take a selfie on their way to Nashville. “We took several days for the drive down and explored the snowy forests in Virginia, a cider brewery in Asheville, North Carolina, and hiking trails in the Smoky Mountains,” Elizabeth wrote. “This picture is from our motel in the Smokies, which were beautiful even in winter.”

56 ASBMB TODAY MAY 2020 PERSPECTIVES

growing up.” “A hamster cage! You got a ham- “The fact that basic scientists are He laughed, and we resuspend- ster!” I said. I was so happy my advice being asked to serve in a patient-fac- ed the cells in freezing media, to be was taken seriously, even if it wasn’t ing role underscores the seriousness stored in liquid nitrogen. “Now, these science-related advice. of the situation at the hospital,” have to go at -80, overnight, then “Yes, her name, or his, I’m not the email stated. “I honestly never they can be moved to LN2 tomor- sure, is Hamilton.” He sent us all a thought that this would happen. row,” he said. photo of the light brown fuzzy critter. The need is real, but so are the risks. Milo then walked into the room Think this through.” Starting to feel familiar and started to chew my backpack My instinct was to tell them all, I occasionally take the bus to straps. “Hey, Milo, don’t eat that “No, don’t do it!” the lab instead of walking, and I am please,” I said. One labmate texted me, “I don’t always the only rider. The driver, “Is that your dog? Show us your know what to do. I want to help, under his mask, usually smiles at me dog,” they said. but I’m also scared. I’m scared that, and recognizes me. “Hello again!” he I turn the laptop to capture Milo, in an emergency situation, I won’t says when I get on. “You done for the careful to hide the pile of still-un- understand what people want me to day?” he says when I see him again packed boxes and miscellaneous mess do.” English is not her first language, on my way home. around my living room. Everyone and I can imagine that urgent orders One time I asked him, “Many “aww”ed at his cute face. coming from mouths hidden beneath other people out today?” When it was my turn to share, masks might make it really hard “Well, including you, I had three I talked about making my new cells to understand what to do and people on the bus today.” lines and trying to get a handle on where to go. One day, a person I recognized the literature. “It’s a little frustrating,” I told her I understood and that from the dog park waved when he I said, “because I’m used to reading she didn’t have to go. saw me walking Milo. We walked papers in my field and understanding together, staying six or more feet them. I keep having to read these ‘You lazy slacker’ apart. His store has closed. We let our over and over again, not getting it.” I really like my new lab, as weird dogs play. “You know, text me or call me as things have been, and want to do a I thought of my friends in New with any questions,” my boss said. good job here. York and how our lives are extremely “Ideally you’d be reading in lab and “I’m not doing my best, though,” different now. People were dying could just pop into my office with I said, on the phone to my sister. “I and hospitals overflowing there. And questions.” I left the lab meeting feel so stupid. I’m not understanding there I was in a sunny park, chatting feeling less alone. stuff quickly and not focusing. I read almost normally. After the meeting, I saw that papers and just nothing sticks. I’m Milo had, in fact, chewed through really afraid of disappointing my new Lab community from afar my backpack strap. I tied it back to- boss! And I don’t want my boss to On our lab’s first Zoom meeting, gether, temporarily fixing it, but the regret hiring me. You know they say we talked about how the lab will knot came out the other day. when Isaac Newton was sent home to probably have to close soon, but quarantine during the plague, that’s again, none of us knew when. It felt Fear from a distance when he discovered some basic phys- a little like “Goodnight, Wesley, sleep On April 2, my former labmates ics laws, right? I’m such a disappoint- well. I’ll most likely kill you in the at Columbia texted about an email ment to science!” morning,” from “The Princess Bride.” they had received from a professor My sister, always able to see After our PI told us what he was involved in coordinating the universi- through bullshit, replied, “You working on, he said, “Also, I’m basi- ty’s response to COVID-19. It asked mean you aren’t doing your best in cally home-schooling my daughter. for volunteers to transport patients a new city alone in the middle of an I’m with her from 8 to 12, and my at the hospital. There are too many unprecedented global pandemic and wife is with her from 12 to 4. And patients, and the hospital can’t afford economic disaster? I can’t believe it! also, I have to finish this hamster cage to let doctors and healthcare workers God, Beth. You lazy slacker.” I started building.” walk away to transport patients. It made me feel a little better.

MAY 2020 ASBMB TODAY 57 PERSPECTIVES

Far from danger? wonder if I might be one of them if If we survive this, then one day My friends in New York are it gets bad here. I wonder who would this empty lab in the empty building facing a terrifying situation that I am take care of Milo if I had to go to the will be my work home, this apart- not a part of. I feel like I abandoned hospital. ment building with its closed rooms them during this impossibly hard I am, technically, quite alone. near the closed park in this city full time. I text my sympathy, but it isn’t “I’m sorry you had to move here to of closed stores and restaurants will quite the same. I watch the numbers a new city and then not be allowed really become my home, and these in the news articles climb. to make any friends,” my PI tells people I see occasionally and from Here, I’m waiting for something. me. But, somehow, I don’t feel fully a distance and through screens will Waiting for things to get worse, or alone. become my friends. waiting for things to get better. Wait- Whether it’s the bus driver who ing for the lab to close, and waiting recognizes me, the dog owners who for the lab to reopen. Waiting for wave, the Skype and Zoom calls with the outbreak to hit us like it’s hitting my lab here, or friends in NYC, I Elizabeth Stivison New York, or waiting for proof that feel OK here. An up-in-the-air type (elizabeth.stivison@ our social distancing is saving us of OK, though. gmail.com) is a postdoc at Vanderbilt University from that outcome. I sit here in a type of limbo, with studying inositol signaling I hear more about healthy people my heart and mind open, knowing it and a careers columnist for the ASBMB. dying from the virus in NYC, and I will get both better and worse.

COVID-19 Special Edition Podcast

ASBMB Public Aairs Director Ben Corb interviews members of the community about their expriences with the ongoing pandemic.

asbmb.org/career-resources/online-resources

58 ASBMB TODAY MAY 2020 ESSAY

Is more science the medicine we need to cure the world’s struggling economy? By Maurizio Crestani

s part of “Life in the time of and ChIP-seq experiments. asymptomatic SARS-Cov-2–positive COVID-19,” I reported on the As you have probably heard, the subjects turned out to be much more Asituation in Italy for the ASBMB situation here in Italy is slowly get- numerous than we expected, and they Today website in the middle of the ting better, but we are aware that the may have contributed substantially COVID-19 outbreak in mid-March. pandemic is far from over. Intensive to the spread of the virus. Not that Here I am, some six weeks later, to care units no longer are overcrowded, it’s their fault, of course, but there’s share more thoughts. as they were a few weeks ago. People no doubt we need to identify them in When I first wrote, Italy was are starting to think that we could order to prevent other uncontrolled completely closed, except phar- gradually return to our jobs. As I waves of the infection. macies, hospitals and food stores. Mi- write this in early May, the lockdown We also need to verify who has lano, usually a very active and alive is being partially lifted. been exposed to the virus and has city, was like a desert. My university Nonetheless, we must be cau- developed antibodies, although we was closed; teaching, meetings, exams tious, as we have learned that this still don’t know whether we develop and thesis defense were being done virus is a tricky one: We have found immunity after infection. And of via teleconferencing. I was working out that people who are positive for course, we need medications and a from home, and my research team the virus but show no symptoms vaccine. In Italy, studies are underway was using the time to analyze data will be a real problem. According to test different therapeutic options, from our experiments, running bio- to some studies performed in small such as infusion of plasma from informatic analyses of our RNA-seq communities in selected areas of Italy, hyperimmune subjects who were pre- LUCA VOLPI / WIKIMEDIA COMMONS VOLPI LUCA

The facade of the headquarters of the Università degli Studi di Milano, as seen from the garden of Largo Richini.

MAY 2020 ASBMB TODAY 59 ESSAY

viously infected by the virus and were and loss of jobs around the world research funding. The members of successfully cured, an antibody against could be investing in fundamental the ASBMB have the energy, the IL-6 used in rheumatoid arthritis, research.Basic science is the engine resources and the contacts to start antivirals for Ebola and more. We are that moves the pipeline of innova- this initiative. Scientists around the also testing vaccines in collaboration tion and new technological devel- world should subscribe to a manifes- with other countries. One vaccine opments, and we need new ideas for to calling for funding fundamental seems quite promising and, if safe and creating jobs and helping to fight the science as an investment in the efficacious, it will be produced by a big economic depression resulting from planet’s future and a prosperous pharmaceutical company adopting a the pandemic. economy that could benefit societies not-for-profit model. I worry that politicians may at all levels. Because, after all, we are In an unpredictable emergency, forget or underestimate the impor- one world. science and scientists are doing their tance of investing in basic science in I’d like to hear from my ASBMB best to find solutions to benefit the any field, from biomedical research colleagues about this proposal. We entire community. We hope the new to engineering to physics. For this must be creative and take steps to knowledge stemming from this effort reason, I think we should launch a realize the dream of a better world. will be available not only for one call for increased funding in scien- Science can certainly help do so. country but worldwide. tific research, as an investment to Cheers from Milano! We must also face another chal- counteract the financial crisis in all lenge: The world economy has been countries. Maurizio Crestani (maurizio. [email protected]) is a struck hard by a tiny microorganism, The American Society for Bio- professor of biochemistry at and that financial crisis is a threat for chemistry and Molecular Biology the Università degli Studi di Milano. His research focuses the near future. I am not an econo- could help start a planetary ini- on epigenetic regulation of mist, but I am sure that one recipe tiative aimed at convincing politi- adipose tissue physiology in the context of obesity and to help fight the economic crisis cians and governments to increase Type 2 diabetes.

PROMOTING RESEARCH OPPORTUNITIES FOR LATIN AMERICAN BIOCHEMISTS

The Promoting Research Opportunities for Latin American Biochemists (PROLAB) program allows Latin American graduate students and postdoctoral fellows to spend up to six months in U.S. or Canadian laboratories.

Apply at asbmb.org/career-resources/ awards-grants-fellowships/prolab

60 ASBMB TODAY MAY 2020 CLASSIFIEDS

Benjamin Goldberg Professor and Head of the Assistant Professor - Immunology-Immune Department of Biochemistry and Molecular Signaling, University of Nebraska-Lincoln Genetics Member Company, UIC College of Department of Biochemistry Medicine

The University of Illinois College of Medicine invites nominations Within the center, UNL is seeking applicants for a nine-month and applications for the position of Benjamin Goldberg Profes- (academic year) tenure-leading Assistant Professor faculty sor and Head of the Department of Biochemistry and Molecu- position (75% research and 25% teaching) in the Department lar Genetics (BMG). BMG has 20 faculty members with active of Biochemistry addressing the sequence of immune signaling research in a variety of areas including biochemistry, epigenetics, events that are activated in individual cells following the structural biology, gene expression, and signaling. BMG faculty association of cell surface molecules during cell-cell contact employ model organisms and innovative genetic and biochemi- and the relationships to the detection of soluble molecules of cal approaches to study fundamental mechanisms, and perform either host or pathogen origin. The ideal candidate will also translational research to treat human diseases. In this role, the have expertise in computational immunology that would deploy new Head of BMG will have the opportunity to build upon the system-wide quantitative data to probe immune-system functions strengths and catalyze the expansion of the Department. using metabolomics, proteomics and/or transcriptomics. The deployment of such technologies would be expected to provide careers.asbmb.org/job/benjamin-goldberg-professor-and- novel integrative analyses that would transform such data into head-of-the-department-of-biochemistry-and-molecular- valuable biological insights of immune system function. genetics/53242179 careers.asbmb.org/job/assistant-professor-immunology- immune-signaling/53738920/

Instructor - Physical Chemistry Postdoctoral Research Associate Tennessee Technological University (Glycobiology/Biochemistry/Microbiology) Department of Chemistry University of Kentucky

The Department of Chemistry at Tennessee Tech University invites The Department of Molecular and Cellular Biochemistry at the applications for an Instructor of Physical Chemistry position University of Kentucky is recruiting post-doctoral fellows to study available at Tennessee Tech University. The department offers an structure, biosynthesis and function of cell wall in pathogenic ACS-certified undergraduate degree as well as a master’s degree streptococci: Streptococcus pyogenes, Streptococcus mutans and in chemistry and will begin transitioning to a new, state-of-the- Streptococcus agalactiae. These projects will utilize cutting-edge art science building during the 2020-2021 academic year. This is microscopy techniques, protein expression and purification, GC- a one-year temporary, non-tenure-track appointment to begin MS analysis of cell wall composition, construction of bacterial August 1, 2020. mutants and molecular biological and biochemical techniques (PCR, cloning, western blot, etc). careers.asbmb.org/job/instructor-physical- chemistry/53707946/ careers.asbmb.org/job/postdoctoral-research-associate- glycobiologybiochemistrymicrobiology/53430309/

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