CDK2AP1/DOC-1 is a bona fide subunit of the Mi-2/NuRD complex.
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.
Mesh Terms:
Animals, Cell Line, Hela Cells, Humans, Immunoprecipitation, Mass Spectrometry, Mi-2 Nucleosome Remodeling and Deacetylase Complex, Mice, Microscopy, Confocal, Microscopy, Fluorescence, NIH 3T3 Cells, Protein Subunits, Tumor Suppressor Proteins
Animals, Cell Line, Hela Cells, Humans, Immunoprecipitation, Mass Spectrometry, Mi-2 Nucleosome Remodeling and Deacetylase Complex, Mice, Microscopy, Confocal, Microscopy, Fluorescence, NIH 3T3 Cells, Protein Subunits, Tumor Suppressor Proteins
Mol Biosyst
Date: Sep. 01, 2010
PubMed ID: 20523938
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