Am J Physiol Cell Physiol AJP: Heart and Circulatory Physiology
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Am J Physiol Cell Physiol 284: C738-C748, 2003. First published November 20, 2002; doi:10.1152/ajpcell.00389.2002
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Vol. 284, Issue 3, C738-C748, March 2003

Transcriptional regulation of the type I myosin heavy chain gene in denervated rat soleus

K. A. Huey, F. Haddad, A. X. Qin, and K. M. Baldwin

Department of Physiology and Biophysics, University of California, Irvine, California 92697

Denervation (DEN) of rat soleus is associated with a decreased expression of slow type I myosin heavy chain (MHC) and an increased expression of the faster MHC isoforms. The molecular mechanisms behind these shifts remain unclear. We first investigated endogenous transcriptional activity of the type I MHC gene in normal and denervated soleus muscles via pre-mRNA analysis. Our results suggest that the type I MHC gene is regulated via transcriptional processes in the denervated soleus. Deletion and mutational analysis of the rat type I MHC promoter was then used to identify cis elements or regions of the promoter involved in this response. DEN significantly decreased in vivo activity of the -3,500, -2,500, -914, -408, -299, and -215 bp type I MHC promoters, relative to the alpha -skeletal actin promoter. In contrast, normalized -171 promoter activity was unchanged. Mutation of the beta e3 element (-214/-190) in the -215 promoter and deletion of this element (-171 promoter) blunted type I downregulation with DEN. In contrast, beta e3 mutation in the -408 promoters was not effective in attenuating the DEN response, suggesting the existence of additional DEN-responsive sites between -408 and -215. Western blotting and gel mobility supershift assays demonstrated decreased expression and DNA binding of transcription enhancer factor 1 (TEF-1) with DEN, suggesting that this decrease may contribute to type I MHC downregulation in denervated muscle.

slow muscle; transcriptional regulation; pre-mRNA; beta e3 DNA regulatory element; denervation


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