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Double strand DNA break repair (DSBR) comprises multiple pathways. A subset of DSBR pathways, including single strand annealing, involve intermediates with 3' non-homologous tails that must be removed to complete repair. In Saccharomyces cerevisiae, Rad1-Rad10 is the structure-specific endonuclease that cleaves the tails in 3' non-homologous tail removal (3' NHTR). Rad1-Rad10 is also an essential component of the nucleotide excision repair (NER) pathway. In both cases, Rad1-Rad10 requires protein partners for recruitment to the relevant DNA intermediate. Msh2-Msh3 and Saw1 recruit Rad1-Rad10 in 3' NHTR; Rad14 recruits Rad1-Rad10 in NER. We created two rad1 separation-of-function alleles, rad1R203A,K205A and rad1R218A; both are defective in 3' NHTR but functional in NER. In vitro, rad1R203A,K205A was impaired at multiple steps in 3' NHTR. The rad1R218A in vivo phenotype resembles that of msh2- or msh3-deleted cells; recruitment of rad1R218A-Rad10 to recombination intermediates is defective. Interactions among rad1R218A-Rad10 and Msh2-Msh3 and Saw1 are altered and rad1R218A-Rad10 interactions with RPA are compromised. We propose a model in which Rad1-Rad10 is recruited and positioned at the recombination intermediate through interactions, between Saw1 and DNA, Rad1-Rad10 and Msh2-Msh3, Saw1 and Msh2-Msh3 and Rad1-Rad10 and RPA. When any of these interactions is altered, 3' NHTR is impaired.

Titel:	Coordination of Rad1-Rad10 interactions with Msh2-Msh3, Saw1 and RPA is essential for functional 3' non-homologous tail removal.
Autor/in / Beteiligte Person:	Eichmiller, R ; Medina-Rivera, M ; DeSanto, R ; Minca, E ; Kim, C ; Holland, C ; Seol, JH ; Schmit, M ; Oramus, D ; Smith, J ; Gallardo, IF ; Finkelstein, IJ ; Lee, SE ; Surtees, JA
Zeitschrift:	Nucleic acids research, Jg. 46 (2018-06-01), Heft 10, S. 5075-5096
Veröffentlichung:	1992- : Oxford : Oxford University Press ; <i>Original Publication</i>: London, Information Retrieval ltd., 2018
Medientyp:	academicJournal
ISSN:	1362-4962 (electronic)
DOI:	10.1093/nar/gky254
Schlagwort:	DNA Breaks, Double-Stranded DNA End-Joining Repair DNA Repair Enzymes genetics DNA-Binding Proteins genetics Endonucleases genetics MutS Homolog 2 Protein metabolism MutS Homolog 3 Protein genetics MutS Homolog 3 Protein metabolism Mutation Protein Interaction Mapping Replication Protein A genetics Saccharomyces cerevisiae radiation effects Saccharomyces cerevisiae Proteins genetics Single-Strand Specific DNA and RNA Endonucleases genetics Ultraviolet Rays DNA Repair Enzymes metabolism DNA-Binding Proteins metabolism Endonucleases metabolism Replication Protein A metabolism Saccharomyces cerevisiae genetics Saccharomyces cerevisiae Proteins metabolism Single-Strand Specific DNA and RNA Endonucleases metabolism
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't Language: English [Nucleic Acids Res] 2018 Jun 01; Vol. 46 (10), pp. 5075-5096. MeSH Terms: DNA Repair Enzymes / metabolism ; DNA-Binding Proteins / metabolism ; Endonucleases / metabolism ; Replication Protein A / metabolism ; Saccharomyces cerevisiae / genetics ; Saccharomyces cerevisiae Proteins / metabolism ; Single-Strand Specific DNA and RNA Endonucleases / *metabolism ; DNA Breaks, Double-Stranded ; DNA End-Joining Repair ; DNA Repair Enzymes / genetics ; DNA-Binding Proteins / genetics ; Endonucleases / genetics ; MutS Homolog 2 Protein / metabolism ; MutS Homolog 3 Protein / genetics ; MutS Homolog 3 Protein / metabolism ; Mutation ; Protein Interaction Mapping ; Replication Protein A / genetics ; Saccharomyces cerevisiae / radiation effects ; Saccharomyces cerevisiae Proteins / genetics ; Single-Strand Specific DNA and RNA Endonucleases / genetics ; Ultraviolet Rays References: Microbiol Mol Biol Rev. 1999 Jun;63(2):349-404. (PMID: 10357855) ; Biochem J. 2005 Oct 15;391(Pt 2):325-33. (PMID: 15975089) ; J Mol Biol. 2006 Jul 14;360(3):523-36. 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(PMID: 19260026) Grant Information: R01 GM071011 United States GM NIGMS NIH HHS; R01 GM120554 United States GM NIGMS NIH HHS; R01 GM124141 United States GM NIGMS NIH HHS; R25 GM095459 United States GM NIGMS NIH HHS Substance Nomenclature: 0 (DNA-Binding Proteins) ; 0 (MSH3 protein, S cerevisiae) ; 0 (MutS Homolog 3 Protein) ; 0 (RFA1 protein, S cerevisiae) ; 0 (Replication Protein A) ; 0 (Saccharomyces cerevisiae Proteins) ; 0 (Saw1 protein, S cerevisiae) ; EC 3.1.- (Endonucleases) ; EC 3.1.- (RAD1 protein, S cerevisiae) ; EC 3.1.30.1 (RAD10 protein, S cerevisiae) ; EC 3.1.30.1 (Single-Strand Specific DNA and RNA Endonucleases) ; EC 3.6.1.3 (MSH2 protein, S cerevisiae) ; EC 3.6.1.3 (MutS Homolog 2 Protein) ; EC 6.5.1.- (DNA Repair Enzymes) Entry Date(s): Date Created: 20180417 Date Completed: 20190822 Latest Revision: 20221102 Update Code: 20240513 PubMed Central ID: PMC6007489

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Coordination of Rad1-Rad10 interactions with Msh2-Msh3, Saw1 and RPA is essential for functional 3' non-homologous tail removal.