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.
Department of Physiological Chemistry and Cancer Genomics Centre, University Medical Center Utrecht, Utrecht, The Netherlands.
Cornelia G
CG
Spruijt
Stefanie J J
SJ
Bartels
Arie B
AB
Brinkman
Jorrit V
JV
Tjeertes
Ina
I
Poser
Hendrik G
HG
Stunnenberg
Michiel
M
Vermeulen
D000818:Animals; D002460:Cell Line; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex / Q000737:chemistry* / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D021122:Protein Subunits; D025521:Tumor Suppressor Proteins / Q000737:chemistry* / Q000378:metabolism*
C096151:CDK2AP1 protein, human; D021122:Protein Subunits; D025521:Tumor Suppressor Proteins; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex