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J. Biol. Chem., Vol. 278, Issue 38, 36603-36610, September 19, 2003

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hMusTRD11 Represses MEF2 Activation of the Troponin I Slow Enhancer^*

Patsie Polly , Leila M. Haddadi, Laura L. Issa , Nanthakumar Subramaniam, Stephen J. Palmer, Enoch S. E. Tay and Edna C. Hardeman ¶

From the Muscle Development Unit, Children's Medical Research Institute, Wentworthville, New South Wales 2145, Australia

The novel transcription factor hMusTRD11 (human muscle TFII-I repeat domain-containing protein 11; previously named MusTRD1; O'Mahoney, J. V., Guven, K. L., Lin, J., Joya, J. E., Robinson, C. S., Wade, R. P., and Hardeman, E. C. (1998) Mol. Cell. Biol. 18, 6641–6652) was identified in a yeast one-hybrid screen as a protein that binds within an upstream enhancer-containing region of the skeletal muscle-specific gene, TNNI1 (human troponin I slow; hTnI_slow). It has been proposed that hMusTRD11 may play an important role in fiber-specific muscle gene expression by virtue of its ability to bind to an Inr-like element (nucleotides –977 to –960) within the hTnI_slow upstream enhancer-containing region that is necessary for slow fiber-specific expression. In this study we demonstrate that both MEF2C, a known regulator of slow fiber-specific genes, and hMusTRD11 regulate hTnI_slow through the Inr-like element. Co-transfection assays in C2C12 cells and Cos-7 cells demonstrate that hMusTRD11 represses hTnI_slow transcription and prevents MEF2C-mediated activation of hTnI_slow transcription. Gel shift analysis shows that hMusTRD11 can abrogate MEF2C binding to its cognate site in the hTnI_slow enhancer. Glutathione S-transferase pull-down assays demonstrate that hMusTRD11 can interact with both MEF2C and the nuclear receptor co-repressor. The data support the role of hMusTRD11 as a repressor of slow fiber-specific transcription through mechanisms involving direct interactions with MEF2C and the nuclear receptor co-repressor.

Received for publication, December 17, 2002 , and in revised form, June 13, 2003.

^* This work was supported by an National Health and Medical Research Council grant (to E. C. H.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Recipient of a Howard Florey Centenary fellowship from the National Health and Medical Research Council of Australia.

Present address: Victor Chang Cardiac Research Institute, Darlinghurst, NSW 2010, Australia.

^¶ To whom correspondence should be addressed: Muscle Development Unit, Children's Medical Research Inst., Locked Bag 23, Wentworthville, NSW 2145, Australia. E-mail: ehardeman{at}cmri.usyd.edu.au.

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