title: Involvement of budding yeast Rad5 in translesion DNA synthesis through physical interaction with Rev1.
abstract: DNA damage tolerance (DDT) is responsible for genomic stability and cell viability by bypassing the replication block. In Saccharomyces cerevisiae DDT employs two parallel branch pathways to bypass the DNA lesion, namely translesion DNA synthesis (TLS) and error-free lesion bypass, which are mediated by sequential modifications of PCNA. Rad5 has been placed in the error-free branch of DDT because it contains an E3 ligase domain required for PCNA polyubiquitination. Rad5 is a multi-functional protein and may also play a role in TLS, since it interacts with the TLS polymerase Rev1. In this study we mapped the Rev1-interaction domain in Rad5 to the amino acid resolution and demonstrated that Rad5 is indeed involved in TLS possibly through recruitment of Rev1. Genetic analyses show that the dual functions of Rad5 can be separated and reconstituted. Crystal structure analysis of the Rad5-Rev1 interaction reveals a consensus RFF motif in the Rad5 N-terminus that binds to a hydrophobic pocket within the C-terminal domain of Rev1 that is highly conserved in eukaryotes. This study indicates that Rad5 plays a critical role in pathway choice between TLS and error-free DDT.
authors.affiliation: College of Life Sciences, Capital Normal University, Beijing 100048, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Xu
authors.affiliation: College of Life Sciences, Capital Normal University, Beijing 100048, China.
authors.forename: Aiyang
authors.initials: A
authors.lastname: Lin
authors.affiliation: Center for Structure Biology, School of Life Science, Tsinghua University, Beijing 100084, China.
authors.forename: Cuiyan
authors.initials: C
authors.lastname: Zhou
authors.affiliation: Department of Microbiology and Immunology, University of Saskatchewan, Saskatoon, SK S7N 5E5, Canada.
authors.forename: Susan R
authors.initials: SR
authors.lastname: Blackwell
authors.affiliation: College of Life Sciences, Capital Normal University, Beijing 100048, China.
authors.forename: Yiran
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Life Sciences, Capital Normal University, Beijing 100048, China.
authors.forename: Zihao
authors.initials: Z
authors.lastname: Wang
authors.affiliation: College of Life Sciences, Capital Normal University, Beijing 100048, China.
authors.forename: Qianqian
authors.initials: Q
authors.lastname: Feng
authors.affiliation: Center for Structure Biology, School of Life Science, Tsinghua University, Beijing 100084, China.
authors.forename: Ruifang
authors.initials: R
authors.lastname: Guan
authors.affiliation: Department of Microbiology and Immunology, University of Saskatchewan, Saskatoon, SK S7N 5E5, Canada.
authors.forename: Michelle D
authors.initials: MD
authors.lastname: Hanna
authors.affiliation: Center for Structure Biology, School of Life Science, Tsinghua University, Beijing 100084, China zhucheng_chen@tsinghua.edu.cn.
authors.forename: Zhucheng
authors.initials: Z
authors.lastname: Chen
authors.affiliation: College of Life Sciences, Capital Normal University, Beijing 100048, China Department of Microbiology and Immunology, University of Saskatchewan, Saskatoon, SK S7N 5E5, Canada wei.xiao@usask.ca.
authors.forename: Wei
authors.initials: W
authors.lastname: Xiao
mesh_terms: D000595:Amino Acid Sequence; D004249:DNA Damage; D004265:DNA Helicases / Q000737:chemistry / Q000378:metabolism*; D004261:DNA Replication; D004843:Epistasis, Genetic; D008958:Models, Molecular; D009154:Mutation; D009713:Nucleotidyltransferases / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D004718:Saccharomycetales / Q000235:genetics* / Q000378:metabolism*
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D009713:Nucleotidyltransferases; D004265:DNA Helicases