The microphthalmia-associated transcription factor (MITF) requires SWI/SNF enzymes to activate melanocyte specific genes

doi:10.1074/jbc.M512052200

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Papers In Press, published online ahead of print April 28, 2006
J. Biol. Chem, 10.1074/jbc.M512052200
Submitted on November 8, 2005
Revised on April 13, 2006
Accepted on April 28, 2006

The microphthalmia-associated transcription factor (MITF) requires SWI/SNF enzymes to activate melanocyte specific genes

Ivana L. de la Serna, Yasuyuki Ohkawa, Chiduru Higashi, Chaitali Dutta, Jules Osias, Naveen Kommajosyula, Taro Tachibana, and Anthony N. Imbalzano

Biochemistry and Cancer Biology, Medical University of Ohio, Toledo, OH 43614

Corresponding Author: idelaserna{at}meduohio.edu

The Microphthalmia Transcription Factor (Mitf) activates melanocyte specific gene expression, is critical for survival and proliferation of melanocytes during development, and has been described as an oncogene in malignant melanoma. SWI/SNF complexes are ATP dependent chromatin remodeling enzymes that play a role in many developmental processes. To determine the requirement for SWI/SNF enzymes in melanocyte differentiation, we introduced Mitf into fibroblasts that inducibly express dominant negative versions of the SWI/SNF ATPases, BRM or BRG1. These dominant negative SWI/SNF components have been shown to inhibit gene activation events that normally require SWI/SNF enzymes. We found that Mitf-mediated activation of a subset of endogenous melanocyte specific genes required SWI/SNF enzymes but that cell cycle regulation occurred independently of SWI/SNF function. Activation of tyrosinase related protein 1 (Trp1), a melanocyte specific gene, correlated with SWI/SNF dependent changes in chromatin accessibility at the endogenous locus. Both BRG1 and Mitf could be localized to the Trp1 and tyrosinase promoters by chromatin immunoprecipitation (ChIP), while immunofluorescence and immunoprecipitation experiments indicate that Mitf and BRG1 co-localized in the nucleus and physically interacted. Together these results suggest that Mitf can recruit SWI/SNF enzymes to melanocyte specific promoters for the activation of gene expression via induced changes in chromatin structure at endogenous loci.

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