title: In vivo regulation of beta-MHC gene in rodent heart: role of T3 and evidence for an upstream enhancer.
abstract: Cardiac beta-myosin heavy chain (beta-MHC) gene expression is mainly regulated through transcriptional processes. Although these results are based primarily on in vitro cell culture models, relatively little information is available concerning the interaction of key regulatory factors thought to modulate MHC expression in the intact rodent heart. Using a direct gene transfer approach, we studied the in vivo transcriptional activity of different-length beta-MHC promoter fragments in normal control and in altered thyroid states. The test beta-MHC promoter was fused to a firefly luciferase reporter gene, whereas the control alpha-MHC promoter was fused to the Renilla luciferase reporter gene and was used to account for variations in transfection efficiency. Absolute reporter gene activities showed that beta- and alpha-MHC genes were individually and reciprocally regulated by thyroid hormone. The beta-to-alpha ratios of reporter gene expression demonstrated an almost threefold larger beta-MHC gene expression in the longest than in the shorter promoter fragments in normal control animals, implying the existence of an upstream enhancer. A mutation in the putative thyroid response element of the -408-bp beta-MHC promoter construct caused transcriptional activity to drop to null. When studied in the -3, 500-bp beta-MHC promoter, construct activity was reduced ( approximately 100-fold) while thyroid hormone responsiveness was retained. These findings suggest that, even though the bulk of the thyroid hormone responsiveness of the gene is contained within the first 215 bp of the beta-MHC promoter sequence, the exact mechanism of triiodothyronine (T3) action remains to be elucidated.
authors: Department of Physiology and Biophysics, University of California, Irvine, California 92697, USA.
authors: C E
authors: CE
authors: Wright
authors: F
authors: F
authors: Haddad
authors: A X
authors: AX
authors: Qin
authors: P W
authors: PW
authors: Bodell
authors: K M
authors: KM
authors: Baldwin
mesh_terms: D000818:Animals; D001517:Coleoptera / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D006321:Heart / Q000502:physiology / Q000503:physiopathology; D006980:Hyperthyroidism / Q000503:physiopathology*; D007037:Hypothyroidism / Q000503:physiopathology*; D008156:Luciferases / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000096:biosynthesis / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011441:Propylthiouracil; D051381:Rats; D017207:Rats, Sprague-Dawley; D013961:Thyroid Gland / Q000502:physiology*; D014158:Transcription, Genetic; D014284:Triiodothyronine / Q000494:pharmacology / Q000502:physiology*
chemical_list: D014284:Triiodothyronine; D011441:Propylthiouracil; D008156:Luciferases; D018995:Myosin Heavy Chains