CDK2AP1/DOC-1 is a bona fide subunit of the Mi-2/NuRD complex

Mol Biosyst. 2010 Sep;6(9):1700-6. doi: 10.1039/c004108d. Epub 2010 Jun 4.

Abstract

The Mi-2/NuRD (NUcleosome Remodeling and histone Deacetylase) chromatin remodeling complex is a large heterogeneous multiprotein complex associated with transcriptional repression. Here we apply a SILAC based quantitative proteomics approach to show that all known Mi-2/NuRD complex subunits co-purify with Cyclin Dependent Kinase 2 Associated Protein1 (CDK2AP1), also known as Deleted in Oral Cancer 1 (DOC-1). DOC-1 displays in vitro binding affinity for methylated DNA as part of the meCpG binding MBD2/NuRD complex. In luciferase reporter assays, DOC-1 is a potent repressor of transcription. Finally, immunofluorescence experiments reveal co-localization between MBD2 and DOC-1 in mouse NIH-3T3 nuclei. Collectively, these results indicate that DOC-1 is a bona fide subunit of the Mi-2/NuRD chromatin remodeling complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • HeLa Cells
  • Humans
  • Immunoprecipitation
  • Mass Spectrometry
  • Mi-2 Nucleosome Remodeling and Deacetylase Complex / chemistry*
  • Mi-2 Nucleosome Remodeling and Deacetylase Complex / metabolism*
  • Mice
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • NIH 3T3 Cells
  • Protein Subunits
  • Tumor Suppressor Proteins / chemistry*
  • Tumor Suppressor Proteins / metabolism*

Substances

  • CDK2AP1 protein, human
  • Protein Subunits
  • Tumor Suppressor Proteins
  • Mi-2 Nucleosome Remodeling and Deacetylase Complex