@article{721c6cb050b1406da3a7b04265889ec6,
title = "XFrom structure to systems: High-resolution, quantitative genetic analysis of RNA polymerase II",
abstract = "RNA polymerase II (RNAPII) lies at the core of dynamic control of gene expression. Using 53 RNAPII point mutants, we generated a point mutant epistatic miniarray profile (pE-MAP) comprising ∼60,000 quantitative genetic interactions in Saccharomyces cerevisiae. This analysis enabled functional assignment of RNAPII subdomains and uncovered connections between individual regions and other protein complexes. Using splicing microarrays and mutants that alter elongation rates in vitro, we found an inverse relationship between RNAPII speed and in vivo splicing efficiency. Furthermore, the pE-MAP classified fast and slow mutants that favor upstream and downstream start site selection, respectively. The striking coordination of polymerization rate with transcription initiation and splicing suggests that transcription rate is tuned to regulate multiple gene expression steps. The pE-MAP approach provides a powerful strategy to understand other multifunctional machines at amino acid resolution.",
author = "Hannes Braberg and Huiyan Jin and Moehle, {Erica A.} and Chan, {Yujia A.} and Shuyi Wang and Michael Shales and Benschop, {Joris J.} and Morris, {John H.} and Chenxi Qiu and Fuqu Hu and Tang, {Leung K.} and Fraser, {James S.} and Holstege, {Frank C.P.} and Philip Hieter and Christine Guthrie and Kaplan, {Craig D.} and Krogan, {Nevan J.}",
note = "Funding Information: The authors wish to thank members of the Krogan, Kaplan, and Guthrie labs; David Agard, Brad Cairns, Ada Cheng, C. James Ingles, Paivand Jalalian, Tanja Kortemme, Andrej Sali, and Keith Yamamoto for helpful discussion; Steve Buratowski, Olga Calvo, and Eva Nogales for sharing results prior to publication; Steve Hahn and Mike Hampsey for providing tfg2 and sua7 mutant plasmids, respectively; Colm Ryan for help with statistical analysis; and Ricardo Almeida and Jiewei Xu for assistance with growth assays. This work was supported by grants from QB3 at UCSF and the NIH (R01GM084448, R01GM084279, P50GM081879, and R01GM098101 to N.J.K.; R01GM036659 to Roger D. Kornberg for support of C.D.K. for a portion of this work; R01GM097260 to C.D.K.; R01GM021119 to C.G.; and DP5OD009180 to J.S.F.). C.G. is an ACS Research Professor of Molecular Genetics. E.A.M. was supported by an NSF graduate research fellowship. C.D.K. acknowledges a Helen Hay Whitney Fellowship for the early stages of this work. H.B. was supported by the UCSF Biophysics Graduate Program. N.J.K. is a Searle Scholar and a Keck Young Investigator. The content is solely the responsibility of the authors and does not necessarily represent the official views of any of the acknowledged funding agencies. ",
year = "2013",
month = aug,
day = "15",
doi = "10.1016/j.cell.2013.07.033",
language = "English",
volume = "154",
pages = "X775--788",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "4",
}