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SUMO Modification Enhances p66-Mediated Transcriptional Repression of the Mi-2/NuRD Complex

Institute for Genetics, Justus Liebig University Giessen, Giessen, Germany

Received 9 March 2006/ Returned for modification 22 March 2006/ Accepted 24 March 2006

Human p66 and p66ß are two potent transcriptional repressors that interact with the methyl-CpG-binding domain proteins MBD2 and MBD3. An analysis of the molecular mechanisms mediating repression resulted in the identification of two major repression domains in p66 and one in p66ß. Both p66 and p66ß are SUMO-modified in vivo: p66 at two sites (Lys-30 and Lys-487) and p66ß at one site (Lys-33). Expression of SUMO1 enhanced the transcriptional repression activity of Gal-p66 and Gal-p66ß. Mutation of the SUMO modification sites or using a SUMO1 mutant or a dominant negative Ubc9 ligase resulted in a significant decrease of the transcriptional repression of p66 and p66ß. The Mi-2/NuRD components MBD3, RbAp46, RbAp48, and HDAC1 were found to bind to both p66 and p66ß in vivo. Most of the interactions were not affected by the SUMO site mutations in p66 or p66ß, with two exceptions. HDAC1 binding to p66 was lost in the case of a p66K30R mutant, and RbAp46 binding was reduced in the case of a p66ßK33R mutant. These results suggest that interactions within the Mi-2/NuRD complex as well as optimal repression are mediated by SUMOylation.

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