PT  - JOURNAL ARTICLE
AU  - David Mavor
AU  - Kyle A. Barlow
AU  - Daniel Asarnow
AU  - Yuliya Birman
AU  - Derek Britain
AU  - Weilin Chen
AU  - Evan M. Green
AU  - Lillian R. Kenner
AU  - Bruk Mensa
AU  - Leanna S. Morinishi
AU  - Charlotte Nelson
AU  - Erin M. Poss
AU  - Pooja Suresh
AU  - Ruilin Tian
AU  - Taylor Arhar
AU  - Beatrice Ary
AU  - David Bauer
AU  - Ian Bergman
AU  - Rachel Brunetti
AU  - Cynthia Chio
AU  - Shizhong Dai
AU  - Miles Dickinson
AU  - Susanna Elledge
AU  - Cole Helsell
AU  - Nathan Hendel
AU  - Emily Kang
AU  - Nadja Kern
AU  - Matvei Khoroshkin
AU  - Lisa Kirkemo
AU  - Greyson Lewis
AU  - Kevin Lou
AU  - Wesley Marin
AU  - Alison Maxwell
AU  - Peter McTigue
AU  - Douglas Myers-Turnbull
AU  - Tamas Nagy
AU  - Andrew Natale
AU  - Keely Oltion
AU  - Sergei Pourmal
AU  - Gabriel Reder
AU  - Nicholas Rettko
AU  - Peter Rohweder
AU  - Daniel Schwarz
AU  - Sophia Tan
AU  - Paul Thomas
AU  - Ryan Tibble
AU  - Jason Town
AU  - Kaitlyn Tsai
AU  - Fatima Ugur
AU  - Douglas R. Wassarmann
AU  - Alexander Wolff
AU  - Taia Wu
AU  - Derrick Bogdanoff
AU  - Jennifer Li
AU  - Kurt S. Thorn
AU  - Shane O’Conchúir
AU  - Danielle L. Swaney
AU  - Eric D. Chow
AU  - Hiten Madhani
AU  - Sy Redding
AU  - Daniel N. Bolon
AU  - Tanja Kortemme
AU  - Joseph L. DeRisi
AU  - Martin Kampmann
AU  - James S. Fraser
TI  - Research Advance: Extending chemical perturbations of the Ubiquitin fitness landscape in a classroom setting
AID  - 10.1101/139352
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 139352
4099  - http://biorxiv.org/content/early/2017/05/17/139352.short
4100  - http://biorxiv.org/content/early/2017/05/17/139352.full
AB  - Although the primary protein sequence of ubiquitin (Ub) is extremely stable over evolutionary time, it is highly tolerant to mutation during selection experiments performed in the laboratory. We have proposed that this discrepancy results from the difference between fitness under laboratory culture conditions and the selective pressures in changing environments over evolutionary time scales. Building on our previous work (Mavor et al 2016), we used deep mutational scanning to determine how twelve new chemicals (3-Amino-1,2,4-triazole, 5-fluorocytosine, Amphotericin B, CaCl2, Cerulenin, Cobalt Acetate, Menadione, Nickel Chloride, p-fluorophenylalanine, Rapamycin, Tamoxifen, and Tunicamycin) reveal novel mutational sensitivities of ubiquitin residues. We found sensitization of Lys63 in eight new conditions. In total, our experiments have uncovered a sensitizing condition for every position in Ub except Ser57 and Gln62. By determining the Ubiquitin fitness landscape under different chemical constraints, our work helps to resolve the inconsistencies between deep mutational scanning experiments and sequence conservation over evolutionary timescales.Builds on Mavor D, Barlow KA, Thompson S, Barad BA, Bonny AR, Cario CL, Gaskins G, Liu Z, Deming L, Axen SD, Caceres E, Chen W, Cuesta A, Gate R, Green EM, Hulce KR, Ji W, Kenner LR, Mensa B, Morinishi LS, Moss SM, Mravic M, Muir RK, Niekamp S, Nnadi CI, Palovcak E, Poss EM, Ross TD, Salcedo E, See S, Subramaniam M, Wong AW, Li J, Thorn KS, Conchúir SÓ, Roscoe BP, Chow ED, DeRisi JL, Kortemme T, Bolon DN, Fraser JS. Determination of Ubiquitin Fitness Landscapes Under Different Chemical Stresses in a Classroom Setting. eLife. 2016.Impact Statement We organized a project-based course that used deep mutational scanning in multiple chemical conditions to resolve the inconsistencies between tolerance to mutations in laboratory conditions and sequence conservation over evolutionary timescales.