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dc.contributor.authorHan, Min-Su
dc.contributor.authorKim, Hyo-Jin
dc.contributor.authorWee, Hee-Jun
dc.contributor.authorLim, Kyung-Eun
dc.contributor.authorPark, Na-Rae
dc.contributor.authorBae, Suk-Chul
dc.contributor.authorVan Wijnen, Andre J.
dc.contributor.authorStein, Janet L.
dc.contributor.authorLian, Jane B.
dc.contributor.authorStein, Gary S.
dc.contributor.authorChoi, Je-Yong
dc.date2022-08-11T08:10:56.000
dc.date.accessioned2022-08-23T17:25:51Z
dc.date.available2022-08-23T17:25:51Z
dc.date.issued2010-05-13
dc.date.submitted2011-01-11
dc.identifier.citationJ Cell Biochem. 2010 May;110(1):97-103. <a href="http://dx.doi.org/10.1002/jcb.22516">Link to article on publisher's site</a>
dc.identifier.issn0730-2312 (Linking)
dc.identifier.doi10.1002/jcb.22516
dc.identifier.pmid20225274
dc.identifier.urihttp://hdl.handle.net/20.500.14038/49499
dc.description.abstractCleidocranial dysplasia (CCD) is caused by haploinsufficiency in RUNX2 function. We have previously identified a series of RUNX2 mutations in Korean CCD patients, including a novel R131G missense mutation in the Runt-homology domain. Here, we examine the functional consequences of the RUNX2(R131G) mutation, which could potentially affect DNA binding, nuclear localization signal, and/or heterodimerization with core-binding factor-beta (CBF-beta). Immunofluorescence microscopy and western blot analysis with subcellular fractions show that RUNX2(R131G) is localized in the nucleus. Immunoprecipitation analysis reveals that heterodimerization with CBF-beta is retained. However, precipitation assays with biotinylated oligonucleotides and reporter gene assays with RUNX2 responsive promoters together reveal that DNA-binding activity and consequently the transactivation of potential of RUNX2(R131G) is abrogated. We conclude that loss of DNA binding, but not nuclear localization or CBF-beta heterodimerization, causes RUNX2 haploinsufficiency in patients with the RUNX2(R131G) mutation. Retention of specific functions including nuclear localization and binding to CBF-beta of the RUNX2(R131G) mutation may render the mutant protein an effective competitor that interferes with wild-type function.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=20225274&dopt=Abstract">Link to Article in PubMed</a>
dc.relation.urlhttp://dx.doi.org/10.1002/jcb.22516
dc.subjectAmino Acid Motifs
dc.subjectAmino Acid Substitution
dc.subjectAnimals
dc.subjectCHO Cells
dc.subjectCell Nucleus
dc.subjectCleidocranial Dysplasia
dc.subjectCore Binding Factor Alpha 1 Subunit
dc.subjectCore Binding Factor Alpha 2 Subunit
dc.subjectCore Binding Factor beta Subunit
dc.subjectCricetinae
dc.subjectCricetulus
dc.subjectDNA
dc.subjectHela Cells
dc.subjectHumans
dc.subjectMutant Proteins
dc.subjectMutation
dc.subjectProtein Binding
dc.subjectProtein Multimerization
dc.subjectProtein Structure, Tertiary
dc.subjectProtein Transport
dc.subjectTranscriptional Activation
dc.subjectCell Biology
dc.titleThe cleidocranial dysplasia-related R131G mutation in the Runt-related transcription factor RUNX2 disrupts binding to DNA but not CBF-beta
dc.typeJournal Article
dc.source.journaltitleJournal of cellular biochemistry
dc.source.volume110
dc.source.issue1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/stein/16
dc.identifier.contextkey1724056
html.description.abstract<p>Cleidocranial dysplasia (CCD) is caused by haploinsufficiency in RUNX2 function. We have previously identified a series of RUNX2 mutations in Korean CCD patients, including a novel R131G missense mutation in the Runt-homology domain. Here, we examine the functional consequences of the RUNX2(R131G) mutation, which could potentially affect DNA binding, nuclear localization signal, and/or heterodimerization with core-binding factor-beta (CBF-beta). Immunofluorescence microscopy and western blot analysis with subcellular fractions show that RUNX2(R131G) is localized in the nucleus. Immunoprecipitation analysis reveals that heterodimerization with CBF-beta is retained. However, precipitation assays with biotinylated oligonucleotides and reporter gene assays with RUNX2 responsive promoters together reveal that DNA-binding activity and consequently the transactivation of potential of RUNX2(R131G) is abrogated. We conclude that loss of DNA binding, but not nuclear localization or CBF-beta heterodimerization, causes RUNX2 haploinsufficiency in patients with the RUNX2(R131G) mutation. Retention of specific functions including nuclear localization and binding to CBF-beta of the RUNX2(R131G) mutation may render the mutant protein an effective competitor that interferes with wild-type function.</p>
dc.identifier.submissionpathstein/16
dc.contributor.departmentDepartment of Cell Biology
dc.source.pages97-103


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