CDK2AP1/DOC-1 is a bona fide subunit of the Mi-2/NuRD complexThis article is part of the 2010 Molecular BioSystems 'Emerging Investigators' issue: highlighting the work of outstanding young scientists at the chemical- and systems-biology interfaces.Electronic supplementary information (ESI) available: Supplementary Tables 1 and 2. See DOI: 10.1039/c004108d

• Main Authors: Spruijt, Cornelia G, Bartels, Stefanie J. J, Brinkman, Arie B, Tjeertes, Jorrit V, Poser, Ina, Stunnenberg, Hendrik G, Vermeulen, Michiel
• Format: Journal Article
• Language: English
• Published: 01.09.2010
• ISSN: 1742-206X; 1742-2051
• DOI: 10.1039/c004108d

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. Using a variety of approaches including state of the art quantitative mass spectrometry technology we provide compelling evidence that the CDK2AP1/DOC-1 protein is a bona fide subunit of the Mi-2/NuRD chromatin remodeling complex.