Genetic Evidence for the Involvement of Mismatch Repair Proteins, PMS2 and MLH3, in a Late Step of Homologous Recombination
| Autor: | Islam Shamima Keka, Shunichi Takeda, Masataka Tsuda, Raphael Guerois, Maminur Rahman, Hiroyuki Sasanuma, Mohiuddin Mohiuddin, Jessica Andreani, Kousei Yamada, Valérie Borde, Jean-Baptiste Charbonnier |
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| Přispěvatelé: | Kyoto University, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Assemblage moléculaire et intégrité du génome (AMIG), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Dynamique de l'information génétique : bases fondamentales et cancer (DIG CANCER), Institut Curie [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Enveloppe Nucléaire, Télomères et Réparation de l’ADN (INTGEN), Kyoto University [Kyoto], Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Sorbonne Université (SU) |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
G2 Phase
0301 basic medicine congenital hereditary and neonatal diseases and abnormalities DNA Repair Resolvase [SDV]Life Sciences [q-bio] RAD51 homologous recombination MLH3 DNA and Chromosomes Biochemistry Piperazines endonuclease Cell Line 03 medical and health sciences Endonuclease PMS2 Holliday junction Humans DNA Breaks Double-Stranded [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology mutL homolog 3 (MLH3) Molecular Biology mutL homolog 1 (MLH1) Mismatch Repair Endonuclease PMS2 DNA Cruciform 030102 biochemistry & molecular biology biology Chemistry MUS81 Cell Biology Joint Molecules Molecular biology digestive system diseases MutL Proteins 030104 developmental biology Gamma Rays Mutation biology.protein Phthalazines Camptothecin DNA mismatch repair GEN1 Homologous recombination |
| Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, inPress, ⟨10.1074/jbc.RA120.013521⟩ Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2020, ⟨10.1074/jbc.RA120.013521⟩ Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, In press, ⟨10.1074/jbc.RA120.013521⟩ J Biol Chem |
| ISSN: | 0021-9258 1083-351X |
| DOI: | 10.1074/jbc.RA120.013521⟩ |
| Popis: | International audience; Homologous recombination (HR) repairs DNA double-strand breaks using intact homologous sequences as template DNA. Broken DNA and intact homologous sequences form joint molecules (JMs), including Holliday junctions (HJs), as HR intermediates. HJs are resolved to form crossover and noncrossover products. A mismatch repair factor, MLH3 endonuclease produces the majority of crossovers during meiotic HR, but it remains elusive whether mismatch repair factors promote HR in non-meiotic cells. We disrupted genes encoding the MLH3 and PMS2 endonucleases in the human B cell line, TK6, generating null MLH3-/- and PMS2-/- mutant cells. We also inserted point mutations into the endonuclease motif of MLH3 and PMS2 genes, generating endonuclease death MLH3DN/DN and PMS2EK/EK cells. MLH3-/- and MLH3DN/DN cells showed a very similar phenotype, a 2.5 times decrease in the frequency of heteroallelic HR-dependent repair of a restriction-enzyme-induced double-strand breaks. PMS2-/- and PMS2EK/EK cells showed a phenotype very similar to that of the MLH3 mutants. These data indicate that MLH3 and PMS2 promote HR as an endonuclease. The MLH3DN/DN and PMS2EK/EK mutations had an additive effect on the heteroallelic HR. MLH3DN/DN/PMS2EK/EK cells showed normal kinetics of g-irradiation-induced Rad51 foci but a significant delay in the resolution of Rad51 foci and three times decrease in the number of cisplatin-induced sister chromatid exchange (SCE). The ectopic expression of the Gen1 HJ resolvase partially reversed the defective heteroallelic HR of MLH3DN/DN/PMS2EK/EK cells. Taken together, we propose that MLH3 and PMS2 promote HR as endonucleases, most likely by processing JMs in mammalian somatic cells. |
| Databáze: | OpenAIRE |
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