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Am J Physiol Cell Physiol (October 24, 2001). doi:10.1152/ajpcell.00444.2001
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Articles in PresS, published online ahead of print October 22, 2001
Am J Physiol Cell Physiol, 10.1152/ajpcell.00444.2001
Submitted on September 17, 2001
Accepted on October 18, 2001

Functional Overload Increases ß Myosin Heavy Chain Promoter Activity in Rodent Fast Muscle through the proximal MCAT (ße3) Site

Julia M Giger1, Fadia Haddad1, Anqi X Qin1, and Kenneth M Baldwin1*

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

* To whom correspondence should be addressed. E-mail: jmeehan{at}uci.edu.

Functional overload (OL) of the rat plantaris muscle by the removal of the synergistic muscles induces a shift in the myosin heavy chain (MHC) isoform expression profile from the fast isoforms toward the slow type I, or, ß MHC isoform. Different length rat ßMHC promoters were linked to a firefly luciferase reporter gene and injected into control and OL plantaris muscles. Reporter activities of -3500, -914, -408, and -215 bp promoters increased in response to one week of OL. The smallest -171 bp promoter was not responsive to OL. Mutation analyses of putative regulatory elements within the -171 and -408 bp region were performed. The -408 promoters containing mutations of the be1, distal MCAT (ße2), CACC, or A/T-rich (GATA) were still responsive to OL. Only the proximal MCAT (ße3) mutation abolished the OL response. Gel mobility shift assays revealed a significantly higher level of complex formation of the ße3 probe with nuclear protein from OL plantaris compared to control plantaris. These results suggest that the ße3 site functions as a putative OL-responsive element in the rat ßMHC gene promoter.




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