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dc.contributor.authorMiret, Juan J.
dc.contributor.authorMilla, Maria G.
dc.contributor.authorLahue, Robert S.
dc.date2022-08-11T08:10:04.000
dc.date.accessioned2022-08-23T16:54:23Z
dc.date.available2022-08-23T16:54:23Z
dc.date.issued1993-02-15
dc.date.submitted2008-08-15
dc.identifier.citationJ Biol Chem. 1993 Feb 15;268(5):3507-13.
dc.identifier.issn0021-9258 (Print)
dc.identifier.pmid8429025
dc.identifier.urihttp://hdl.handle.net/20.500.14038/42492
dc.description.abstractAn activity present in nuclear extracts of the yeast Saccharomyces cerevisiae binds specifically to oligonucleotides containing DNA mismatches, as judged by a band shift assay. The specificity of this activity for mismatched DNA was confirmed by competition experiments; binding to radiolabeled heteroduplexes was abolished in the presence of excess unlabeled heteroduplex but not when excess unlabeled homoduplex was added. Both T/G and T/- (single base deletion) mispairs were recognized in each of two sequence contexts. Binding was also observed with G/G, G/A, A/C, and T/C mismatches, but recognition of a C/C mispair was very weak. Competition studies with the various mismatches were consistent with the idea that a single activity recognizes all mispairs tested. Extracts from strains mutant in either or both of two putative mismatch recognition functions, MSH2 and MSH3, were also tested. Mismatch-binding activity was present in extracts of msh3- strains but completely absent in msh2- strains. The molecular weight of the major binding protein was estimated by UV cross-linking experiments to be approximately 110 kDa, in good agreement with the size predicted for Msh2 protein (Reenan, R. A. and Kolodner, R. D. (1992) Genetics 132, 963-973).
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=8429025&dopt=Abstract">Link to Article in PubMed</a>
dc.relation.urlhttp://www.jbc.org/content/268/5/3507.long
dc.subject*Base Composition
dc.subjectBase Sequence
dc.subjectBinding, Competitive
dc.subjectCell Nucleus
dc.subjectDNA, Fungal
dc.subjectEscherichia coli
dc.subjectGenes, Bacterial
dc.subjectGenes, Fungal
dc.subjectMolecular Sequence Data
dc.subjectNucleic Acid Heteroduplexes
dc.subjectOligodeoxyribonucleotides
dc.subjectSaccharomyces cerevisiae
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleCharacterization of a DNA mismatch-binding activity in yeast extracts
dc.typeJournal Article
dc.source.journaltitleThe Journal of biological chemistry
dc.source.volume268
dc.source.issue5
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/836
dc.identifier.contextkey579723
html.description.abstract<p>An activity present in nuclear extracts of the yeast Saccharomyces cerevisiae binds specifically to oligonucleotides containing DNA mismatches, as judged by a band shift assay. The specificity of this activity for mismatched DNA was confirmed by competition experiments; binding to radiolabeled heteroduplexes was abolished in the presence of excess unlabeled heteroduplex but not when excess unlabeled homoduplex was added. Both T/G and T/- (single base deletion) mispairs were recognized in each of two sequence contexts. Binding was also observed with G/G, G/A, A/C, and T/C mismatches, but recognition of a C/C mispair was very weak. Competition studies with the various mismatches were consistent with the idea that a single activity recognizes all mispairs tested. Extracts from strains mutant in either or both of two putative mismatch recognition functions, MSH2 and MSH3, were also tested. Mismatch-binding activity was present in extracts of msh3- strains but completely absent in msh2- strains. The molecular weight of the major binding protein was estimated by UV cross-linking experiments to be approximately 110 kDa, in good agreement with the size predicted for Msh2 protein (Reenan, R. A. and Kolodner, R. D. (1992) Genetics 132, 963-973).</p>
dc.identifier.submissionpathoapubs/836
dc.contributor.departmentDepartment of Biochemistry and Molecular Biology
dc.source.pages3507-13


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