title: S. cerevisiae Mre11 recruits conjugated SUMO moieties to facilitate the assembly and function of the Mre11-Rad50-Xrs2 complex.
abstract: Double-strand breaks (DSBs) in chromosomes are the most challenging type of DNA damage. The yeast and mammalian Mre11-Rad50-Xrs2/Nbs1 (MRX/N)-Sae2/Ctp1 complex catalyzes the resection of DSBs induced by secondary structures, chemical adducts or covalently-attached proteins. MRX/N also initiates two parallel DNA damage responses-checkpoint phosphorylation and global SUMOylation-to boost a cell's ability to repair DSBs. However, the molecular mechanism of this SUMO-mediated response is not completely known. In this study, we report that Saccharomyces cerevisiae Mre11 can non-covalently recruit the conjugated SUMO moieties, particularly the poly-SUMO chain. Mre11 has two evolutionarily-conserved SUMO-interacting motifs, Mre11(SIM1) and Mre11(SIM2), which reside on the outermost surface of Mre11. Mre11(SIM1) is indispensable for MRX assembly. Mre11(SIM2) non-covalently links MRX with the SUMO enzymes (E2/Ubc9 and E3/Siz2) to promote global SUMOylation of DNA repair proteins. Mre11(SIM2) acts independently of checkpoint phosphorylation. During meiosis, the mre11(SIM2) mutant, as for mre11S, rad50S and sae2Δ, allows initiation but not processing of Spo11-induced DSBs. Using MRX and DSB repair as a model, our work reveals a general principle in which the conjugated SUMO moieties non-covalently facilitate the assembly and functions of multi-subunit protein complexes.
authors.affiliation: Graduate Program of Biotechnology in Medicine, National Tsing Hua University and National Health Research Institutes, Taiwan Institute of Biotechnology, National Tsing Hua University, Hsinchu 300, Taiwan National Institute of Infectious Diseases and Vaccinology, National Health Research Institute, Miaoli 350, Taiwan Institute of Molecular Biology, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Yu-Jie
authors.initials: YJ
authors.lastname: Chen
authors.affiliation: Institute of Molecular Biology, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Yu-Chien
authors.initials: YC
authors.lastname: Chuang
authors.affiliation: Institute of Molecular Biology, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Chi-Ning
authors.initials: CN
authors.lastname: Chuang
authors.affiliation: Institute of Molecular Biology, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Yun-Hsin
authors.initials: YH
authors.lastname: Cheng
authors.affiliation: Graduate Program of Biotechnology in Medicine, National Tsing Hua University and National Health Research Institutes, Taiwan Institute of Biotechnology, National Tsing Hua University, Hsinchu 300, Taiwan.
authors.forename: Chuang-Rung
authors.initials: CR
authors.lastname: Chang
authors.affiliation: Graduate Program of Biotechnology in Medicine, National Tsing Hua University and National Health Research Institutes, Taiwan National Institute of Infectious Diseases and Vaccinology, National Health Research Institute, Miaoli 350, Taiwan tfwang@gate.sinica.edu.tw.
authors.forename: Chih-Hsiang
authors.initials: CH
authors.lastname: Leng
authors.affiliation: Institute of Molecular Biology, Academia Sinica, Taipei 115, Taiwan tfwang@gate.sinica.edu.tw.
authors.forename: Ting-Fang
authors.initials: TF
authors.lastname: Wang
mesh_terms: D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D004271:DNA, Fungal / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000235:genetics* / Q000378:metabolism; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism; D005090:Exodeoxyribonucleases / Q000235:genetics* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D008540:Meiosis; D008958:Models, Molecular; D010766:Phosphorylation; D011485:Protein Binding; D055503:Protein Multimerization; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D058207:Sumoylation; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics / Q000378:metabolism; D000098843:Ubiquitin-Conjugating Enzyme UBC9; D000099257:Meiotic Recombination Protein SPO11
chemical_list: D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D004706:Endodeoxyribonucleases; D004720:Endonucleases; D005090:Exodeoxyribonucleases; D029701:Saccharomyces cerevisiae Proteins; D044763:Ubiquitin-Conjugating Enzymes; D000098843:Ubiquitin-Conjugating Enzyme UBC9; C086930:RAD50 protein, S cerevisiae; C107849:SAE2 protein, S cerevisiae; C436836:Siz2 protein, S cerevisiae; C086929:XRS2 protein, S cerevisiae; C094191:MRE11 protein, S cerevisiae; C518938:Spo11 protein, S cerevisiae; D000099257:Meiotic Recombination Protein SPO11