Unlocking the steric gate of DNA polymerase η leads to increased genomic instability in Saccharomyces cerevisiae

DNA Repair
Katherine A DoniganRoger Woodgate

Abstract

DNA polymerase η (pol η) is best characterized for its ability to perform accurate and efficient translesion DNA synthesis (TLS) through cyclobutane pyrimidine dimers (CPDs). To ensure accurate bypass the polymerase is not only required to select the correct base, but also discriminate between NTPs and dNTPs. Most DNA polymerases have a conserved "steric gate" residue which functions to prevent incorporation of NMPs during DNA synthesis. Here, we demonstrate that the Phe35 residue of Saccharomyces cerevisiae pol η functions as a steric gate to limit the use of ribonucleotides during polymerization both in vitro and in vivo. Unlike the related pol ι enzyme, wild-type pol η does not readily incorporate NMPs in vitro. In contrast, a pol η F35A mutant incorporates NMPs on both damaged and undamaged DNA in vitro with a high degree of base selectivity. An S.cerevisiae strain expressing pol η F35A (rad30-F35A) that is also deficient for nucleotide excision repair (rad1Δ) and the TLS polymerase, pol ζ (rev3Δ), is extremely sensitive to UV-light. The sensitivity is due, in part, to RNase H2 activity, as an isogenic rnh201Δ strain is roughly 50-fold more UV-resistant than its RNH201(+) counterpart. Interestingly the rad1Δ rev3Δ rad30-F35...Continue Reading

References

Mar 25, 1992·Nucleic Acids Research·D GietzR H Schiestl
Dec 1, 1986·Mutation Research·G DasF Sherman
Nov 9, 1994·Molecular and Cellular Biochemistry·T W Traut
May 9, 2000·Nature·T MatsudaT A Kunkel
Oct 31, 2000·The Journal of Biological Chemistry·M T WashingtonL Prakash
Aug 23, 2001·Molecular Cell·H OhmoriR Woodgate
Jun 5, 2002·Annual Review of Biochemistry·Myron F Goodman
Dec 12, 2002·Proceedings of the National Academy of Sciences of the United States of America·Bjorn Rydberg, John Game
Jan 22, 2004·Nucleic Acids Research·Ho-Sang JeongRobert J Crouch
May 27, 2005·Molecular Cell·Errol C FriedbergRobert P P Fuchs
Sep 28, 2007·Proceedings of the National Academy of Sciences of the United States of America·Wei Yang, Roger Woodgate
Nov 28, 2007·Nucleic Acids Research·Shibani DalalJoann B Sweasy
Jan 2, 2008·Cell Research·Scott D McCulloch, Thomas A Kunkel
Apr 22, 2009·Molecular and Cellular Biology·Laurie ReyJean-Sébastien Hoffmann
Dec 17, 2009·Nucleic Acids Research·Paola FerraroVera Bianchi
Aug 24, 2010·Nature Chemical Biology·Stephanie A Nick McElhinnyThomas A Kunkel
Sep 18, 2010·Journal of Nucleic Acids·Adam D BrownAlexander J R Bishop
Oct 12, 2010·Seminars in Cancer Biology·Mercedes E Arana, Thomas A Kunkel
Jun 28, 2011·Science·Nayun KimSue Jinks-Robertson
Sep 8, 2011·Protein Science : a Publication of the Protein Society·Wei Yang
Jul 18, 2012·The Journal of Experimental Medicine·Bjoern HillerAxel Roers
Aug 7, 2012·Molecular Cell·Justin L SparksPeter M Burgers
Dec 19, 2012·DNA Repair·Anders R ClausenThomas A Kunkel
Jan 12, 2013·DNA Repair·Jang-Eun ChoSue Jinks-Robertson
Jan 29, 2013·Nucleic Acids Research·Hyongi ChonSusana M Cerritelli
Feb 5, 2013·Molecular Cell·Jessica S WilliamsThomas A Kunkel
Apr 24, 2013·The Journal of Cell Biology·Valérie BergoglioJean-Sébastien Hoffmann
Jul 11, 2013·Cold Spring Harbor Perspectives in Biology·Myron F Goodman, Roger Woodgate
Oct 8, 2013·Journal of Molecular Biology·Anne-Sophie BoyerJean-Sébastien Hoffmann
Feb 6, 2014·Mutation Research. Fundamental and Molecular Mechanisms of Mutagenesis·Alexandra VaismanRoger Woodgate
Feb 18, 2014·The Journal of Biological Chemistry·Katherine A DoniganRoger Woodgate

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Citations

Mar 22, 2016·Trends in Biochemical Sciences·Susana M Cerritelli, Robert J Crouch
Apr 21, 2016·Nature Reviews. Molecular Cell Biology·Jessica S WilliamsThomas A Kunkel
Mar 11, 2017·Critical Reviews in Biochemistry and Molecular Biology·Alexandra Vaisman, Roger Woodgate
Jul 5, 2018·Critical Reviews in Biochemistry and Molecular Biology·Alexandra Vaisman, Roger Woodgate
Jul 20, 2019·Infant Mental Health Journal·Andre Dukes, Glen Palm
Sep 16, 2017·Nucleic Acids Research·Andrea F MoonLars C Pedersen
Jun 7, 2019·Mutagenesis·Alexandra BerroyerErik D Larson
Mar 7, 2020·International Journal of Molecular Sciences·Giulia Maria NavaFederico Lazzaro
Jan 1, 2019·Nucleic Acids Research·Katrin KreiselAnders R Clausen
Jan 8, 2016·The Journal of Biological Chemistry·Yan SuF Peter Guengerich
Jan 20, 2021·Critical Reviews in Biochemistry and Molecular Biology·Zhi-Xiong ZhouThomas A Kunkel
Nov 26, 2019·DNA Repair·Susana M Cerritelli, Robert J Crouch

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