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dc.contributor.authorGroothuizen, Flora S.
dc.contributor.authorFish, Alexander
dc.contributor.authorPetoukhov, Maxim V.
dc.contributor.authorReumer, Annet
dc.contributor.authorManelyte, Laura
dc.contributor.authorWinterwerp, Herrie H.K.
dc.contributor.authorMarinus, Martin G.
dc.contributor.authorLebbink, Joyce H.G.
dc.contributor.authorSvergun, Dmitri I.
dc.contributor.authorFriedhoff, Peter
dc.contributor.authorSixma, Titia K.
dc.date2022-08-11T08:08:30.000
dc.date.accessioned2022-08-23T15:57:32Z
dc.date.available2022-08-23T15:57:32Z
dc.date.issued2013-09-01
dc.date.submitted2014-12-23
dc.identifier.citationNucleic Acids Res. 2013 Sep;41(17):8166-81. doi: 10.1093/nar/gkt582. Epub 2013 Jul 1. <a href="http://dx.doi.org/10.1093/nar/gkt582">Link to article on publisher's site</a>
dc.identifier.issn0305-1048 (Linking)
dc.identifier.doi10.1093/nar/gkt582
dc.identifier.pmid23821665
dc.identifier.urihttp://hdl.handle.net/20.500.14038/30219
dc.description.abstractThe process of DNA mismatch repair is initiated when MutS recognizes mismatched DNA bases and starts the repair cascade. The Escherichia coli MutS protein exists in an equilibrium between dimers and tetramers, which has compromised biophysical analysis. To uncouple these states, we have generated stable dimers and tetramers, respectively. These proteins allowed kinetic analysis of DNA recognition and structural analysis of the full-length protein by X-ray crystallography and small angle X-ray scattering. Our structural data reveal that the tetramerization domains are flexible with respect to the body of the protein, resulting in mostly extended structures. Tetrameric MutS has a slow dissociation from DNA, which can be due to occasional bending over and binding DNA in its two binding sites. In contrast, the dimer dissociation is faster, primarily dependent on a combination of the type of mismatch and the flanking sequence. In the presence of ATP, we could distinguish two kinetic groups: DNA sequences where MutS forms sliding clamps and those where sliding clamps are not formed efficiently. Interestingly, this inability to undergo a conformational change rather than mismatch affinity is correlated with mismatch repair.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=23821665&dopt=Abstract">Link to Article in PubMed</a>
dc.rights.urihttp://creativecommons.org/licenses/by-nc/3.0/
dc.subjectAdenosine Triphosphate
dc.subject*Base Pair Mismatch
dc.subjectDNA
dc.subjectEscherichia coli Proteins
dc.subjectModels, Molecular
dc.subjectMutS DNA Mismatch-Binding Protein
dc.subjectProtein Binding
dc.subjectProtein Multimerization
dc.subjectProtein Structure, Tertiary
dc.subjectBiochemistry
dc.subjectMolecular Biology
dc.titleUsing stable MutS dimers and tetramers to quantitatively analyze DNA mismatch recognition and sliding clamp formation
dc.typeJournal Article
dc.source.journaltitleNucleic acids research
dc.source.volume41
dc.source.issue17
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=1467&amp;context=faculty_pubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/faculty_pubs/468
dc.identifier.contextkey6488312
refterms.dateFOA2022-08-23T15:57:32Z
html.description.abstract<p>The process of DNA mismatch repair is initiated when MutS recognizes mismatched DNA bases and starts the repair cascade. The Escherichia coli MutS protein exists in an equilibrium between dimers and tetramers, which has compromised biophysical analysis. To uncouple these states, we have generated stable dimers and tetramers, respectively. These proteins allowed kinetic analysis of DNA recognition and structural analysis of the full-length protein by X-ray crystallography and small angle X-ray scattering. Our structural data reveal that the tetramerization domains are flexible with respect to the body of the protein, resulting in mostly extended structures. Tetrameric MutS has a slow dissociation from DNA, which can be due to occasional bending over and binding DNA in its two binding sites. In contrast, the dimer dissociation is faster, primarily dependent on a combination of the type of mismatch and the flanking sequence. In the presence of ATP, we could distinguish two kinetic groups: DNA sequences where MutS forms sliding clamps and those where sliding clamps are not formed efficiently. Interestingly, this inability to undergo a conformational change rather than mismatch affinity is correlated with mismatch repair.</p>
dc.identifier.submissionpathfaculty_pubs/468
dc.contributor.departmentDepartment of Biochemistry and Molecular Pharmacology
dc.source.pages8166-81


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