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Am J Physiol Cell Physiol (November 20, 2002). doi:10.1152/ajpcell.00389.2002
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Articles in PresS, published online ahead of print November 20, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00389.2002
Submitted on August 27, 2002
Accepted on November 14, 2002

Transcriptional Regulation of the Type I Myosin Heavy Chain Gene in Denervated Rat Soleus

Kimberly A Huey1*, Fadia Haddad1, Anqi X Qin1, and Kenneth M Baldwin1

1 Physiology and Biophysics, University of California, Irvine, Irvine, CA, USA

* To whom correspondence should be addressed. E-mail: kimberly.huey{at}asu.edu.

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 then was used to identify cis elements or regions of the promoter involved in this response. DEN significantly decreased in vivo activity of the -3500,-2500, -914, -408, -299, and -215 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 down-regulation 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 down-regulation in denervated muscle.




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