Characterization of a DNA mismatch-binding activity in yeast extracts
dc.contributor.author | Miret, Juan J. | |
dc.contributor.author | Milla, Maria G. | |
dc.contributor.author | Lahue, Robert S. | |
dc.date | 2022-08-11T08:10:04.000 | |
dc.date.accessioned | 2022-08-23T16:54:23Z | |
dc.date.available | 2022-08-23T16:54:23Z | |
dc.date.issued | 1993-02-15 | |
dc.date.submitted | 2008-08-15 | |
dc.identifier.citation | J Biol Chem. 1993 Feb 15;268(5):3507-13. | |
dc.identifier.issn | 0021-9258 (Print) | |
dc.identifier.pmid | 8429025 | |
dc.identifier.uri | http://hdl.handle.net/20.500.14038/42492 | |
dc.description.abstract | 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). | |
dc.language.iso | en_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.url | http://www.jbc.org/content/268/5/3507.long | |
dc.subject | *Base Composition | |
dc.subject | Base Sequence | |
dc.subject | Binding, Competitive | |
dc.subject | Cell Nucleus | |
dc.subject | DNA, Fungal | |
dc.subject | Escherichia coli | |
dc.subject | Genes, Bacterial | |
dc.subject | Genes, Fungal | |
dc.subject | Molecular Sequence Data | |
dc.subject | Nucleic Acid Heteroduplexes | |
dc.subject | Oligodeoxyribonucleotides | |
dc.subject | Saccharomyces cerevisiae | |
dc.subject | Life Sciences | |
dc.subject | Medicine and Health Sciences | |
dc.title | Characterization of a DNA mismatch-binding activity in yeast extracts | |
dc.type | Journal Article | |
dc.source.journaltitle | The Journal of biological chemistry | |
dc.source.volume | 268 | |
dc.source.issue | 5 | |
dc.identifier.legacycoverpage | https://escholarship.umassmed.edu/oapubs/836 | |
dc.identifier.contextkey | 579723 | |
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.submissionpath | oapubs/836 | |
dc.contributor.department | Department of Biochemistry and Molecular Biology | |
dc.source.pages | 3507-13 |