The cleidocranial dysplasia-related R131G mutation in the runt-related transcription factor RUNX2 disrupts binding to DNA but not CBF-β

Min Su Han, Hyo Jin Kim, Hee Jun Wee, Kyung Eun Lim, Na Rae Park, Suk Chul Bae, Andre J. Van Wijnen, Janet L. Stein, Jane B. Lian, Gary S. Stein, Je Yong Choi

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

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-β (CBF-β). Immunofluorescence microscopy and western blot analysis with subcellular fractions show that RUNX2R131G is localized in the nucleus. Immunoprecipitation analysis reveals that heterodimerization with CBF-β 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 RUNX2R131G is abrogated. We conclude that loss of DNA binding, but not nuclear localization or CBF-β heterodimerization, causes RUNX2 haploinsufficiency in patients with the RUNX2R131G mutation. Retention of specific functions including nuclear localization and binding to CBF-β of the RUNX2R131G mutation may render the mutant protein an effective competitor that interferes with wild-type function.

Original languageEnglish (US)
Pages (from-to)97-103
Number of pages7
JournalJournal of cellular biochemistry
Volume110
Issue number1
DOIs
StatePublished - May 1 2010

Keywords

  • Cleidocranial dysplasia (CCD)
  • Core-binding factor-β (CBF-β)
  • DNA-binding activity
  • Heterodimerization
  • RUNX2
  • RUNX2
  • Subcellular localization
  • Transactivation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Han, M. S., Kim, H. J., Wee, H. J., Lim, K. E., Park, N. R., Bae, S. C., Van Wijnen, A. J., Stein, J. L., Lian, J. B., Stein, G. S., & Choi, J. Y. (2010). The cleidocranial dysplasia-related R131G mutation in the runt-related transcription factor RUNX2 disrupts binding to DNA but not CBF-β. Journal of cellular biochemistry, 110(1), 97-103. https://doi.org/10.1002/jcb.22516