The Shu complex promotes error-free tolerance of alkylation-induced base excision repair products.
Here, we investigate the role of the budding yeast Shu complex in promoting homologous recombination (HR) upon replication fork damage. We recently found that the Shu complex stimulates Rad51 filament formation during HR through its physical interactions with Rad55-Rad57. Unlike other HR factors, Shu complex mutants are primarily sensitive to replicative stress caused by MMS and not to more direct DNA breaks. Here, we uncover a novel role for the Shu complex in the repair of specific MMS-induced DNA lesions and elucidate the interplay between HR and translesion DNA synthesis. We find that the Shu complex promotes high-fidelity bypass of MMS-induced alkylation damage, such as N3-methyladenine, as well as bypassing the abasic sites generated after Mag1 removes N3-methyladenine lesions. Furthermore, we find that the Shu complex responds to ssDNA breaks generated in cells lacking the abasic site endonucleases. At each lesion, the Shu complex promotes Rad51-dependent HR as the primary repair/tolerance mechanism over error-prone translesion DNA polymerases. Together, our work demonstrates that the Shu complex's promotion of Rad51 pre-synaptic filaments is critical for high-fidelity bypass of multiple replication-blocking lesion.
University of Pittsburgh School of Medicine, Department of Microbiology and Molecular Genetics, 5117 Centre Avenue, Pittsburgh, PA 15213, USA.
Stephen K
SK
Godin
University of Pittsburgh School of Medicine, Department of Microbiology and Molecular Genetics, 5117 Centre Avenue, Pittsburgh, PA 15213, USA Tsinghua University School of Medicine, Tsinghua University, Haidian District, Beijing 100084, China.
Zhuying
Z
Zhang
University of Pittsburgh School of Medicine, Department of Microbiology and Molecular Genetics, 5117 Centre Avenue, Pittsburgh, PA 15213, USA.
Benjamin W
BW
Herken
Chromosome Stability Group, Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences (NIEHS), National Institutes of Health, Research Triangle Park, NC 27709, USA.
James W
JW
Westmoreland
University of Pittsburgh School of Medicine, Department of Microbiology and Molecular Genetics, 5117 Centre Avenue, Pittsburgh, PA 15213, USA.
Alison G
AG
Lee
University of Pittsburgh School of Medicine, Department of Microbiology and Molecular Genetics, 5117 Centre Avenue, Pittsburgh, PA 15213, USA.
Michael J
MJ
Mihalevic
Tsinghua University School of Medicine, Tsinghua University, Haidian District, Beijing 100084, China Department of Pharmacology & Chemical Biology, Pittsburgh, PA 15217, USA.
Zhongxun
Z
Yu
Department of Pharmacology & Chemical Biology, Pittsburgh, PA 15217, USA University of South Alabama Mitchell Cancer Institute, 1660 Springhill Avenue, Mobile, AL 36604, USA.
Robert W
0000-0001-7385-3563
RW
Sobol
Chromosome Stability Group, Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences (NIEHS), National Institutes of Health, Research Triangle Park, NC 27709, USA.
Michael A
MA
Resnick
University of Pittsburgh School of Medicine, Department of Microbiology and Molecular Genetics, 5117 Centre Avenue, Pittsburgh, PA 15213, USA karab@pitt.edu.
Kara A
KA
Bernstein
D000225:Adenine / Q000031:analogs & derivatives / Q000378:metabolism; D000478:Alkylation; D002166:Camptothecin / Q000494:pharmacology; D002945:Cisplatin / Q000494:pharmacology; D004249:DNA Damage / Q000235:genetics; D019951:DNA Polymerase beta / Q000378:metabolism; D004260:DNA Repair / Q000187:drug effects; D004271:DNA, Fungal / Q000096:biosynthesis; D004843:Epistasis, Genetic / Q000187:drug effects; D005047:Etoposide / Q000494:pharmacology; D005800:Genes, Fungal; D056426:Genetic Loci; D059765:Homologous Recombination / Q000235:genetics; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006918:Hydroxyurea / Q000494:pharmacology; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D008954:Models, Biological; D009154:Mutation / Q000235:genetics; D059645:Mutation Rate; D011485:Protein Binding / Q000187:drug effects; D011839:Radiation, Ionizing; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism* / Q000528:radiation effects; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014466:Ultraviolet Rays
D004271:DNA, Fungal; C580784:N3-methyladenine; D029701:Saccharomyces cerevisiae Proteins; D005047:Etoposide; D008741:Methyl Methanesulfonate; D006861:Hydrogen Peroxide; D019951:DNA Polymerase beta; D000225:Adenine; D002945:Cisplatin; D006918:Hydroxyurea; D002166:Camptothecin