Human Molecular Genetics Advance Access originally published online on May 29, 2009
Human Molecular Genetics 2009 18(17):3178-3193; doi:10.1093/hmg/ddp256
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
DNMT3B interacts with constitutive centromere protein CENP-C to modulate DNA methylation and the histone code at centromeric regions
1 Department of Biochemistry and Molecular Biology, University of Florida, Box 100245, Gainesville, FL 32610, USA 2 Department of Molecular Genetics and Microbiology, Duke University, 101 Science Dr, Durham, NC 27708, USA 3 Department of Human and General Physiology, P.zza Porta San Donato, 40126 Bologna, Italy 4 Department of Biology, Via Selmi 3, University of Bologna, 40126 Bologna, Italy
* To whom correspondence should be addressed at: Department of Biochemistry and Molecular Biology, University of Florida, Box 100245, 1600 S.W. Archer Road, Gainesville, FL 32610, USA. Tel: +1 3523921810; Fax: +1 3523922953; Email: keithr{at}ufl.edu
Received April 9, 2009; Accepted May 27, 2009
DNA methylation is an epigenetically imposed mark of transcriptional repression that is essential for maintenance of chromatin structure and genomic stability. Genome-wide methylation patterns are mediated by the combined action of three DNA methyltransferases: DNMT1, DNMT3A and DNMT3B. Compelling links exist between DNMT3B and chromosome stability as emphasized by the mitotic defects that are a hallmark of ICF syndrome, a disease arising from germline mutations in DNMT3B. Centromeric and pericentromeric regions are essential for chromosome condensation and the fidelity of segregation. Centromere regions contain distinct epigenetic marks, including dense DNA hypermethylation, yet the mechanisms by which DNA methylation is targeted to these regions remains largely unknown. In the present study, we used a yeast two-hybrid screen and identified a novel interaction between DNMT3B and constitutive centromere protein CENP-C. CENP-C is itself essential for mitosis. We confirm this interaction in mammalian cells and map the domains responsible. Using siRNA knock downs, bisulfite genomic sequencing and ChIP, we demonstrate for the first time that CENP-C recruits DNA methylation and DNMT3B to both centromeric and pericentromeric satellite repeats and that CENP-C and DNMT3B regulate the histone code in these regions, including marks characteristic of centromeric chromatin. Finally, we demonstrate that loss of CENP-C or DNMT3B leads to elevated chromosome misalignment and segregation defects during mitosis and increased transcription of centromeric repeats. Taken together, our data reveal a novel mechanism by which DNA methylation is targeted to discrete regions of the genome and contributes to chromosomal stability.
Present address: Medical College of Georgia, Cancer Center and Department of Biochemistry and Molecular Biology, 1120 15th Street, CN-4123, Augusta, GA 30912, USA. Email: krobertson@mcg.edu.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  S. Gopalakrishnan, B. O. Van Emburgh, J. Shan, Z. Su, C. R. Fields, J. Vieweg, T. Hamazaki, P. H. Schwartz, N. Terada, and K. D. Robertson A Novel DNMT3B Splice Variant Expressed in Tumor and Pluripotent Cells Modulates Genomic DNA Methylation Patterns and Displays Altered DNA Binding Mol. Cancer Res., October 1, 2009; 7(10): 1622 - 1634. [Abstract] [Full Text] [PDF]
|
|