Mechanical overload leads to enlargement of adult striated muscle and distinct changes in both myosin phenotype and contractile properties. Remaining critical questions in adult-stage muscle biology involve determining the DNA regulatory element(s) and nuclear protein factor(s) which transcriptionally induce beta myosin heavy chain expression in mechanical overloaded plantaris muscle and to test their possible role in fiber specific expression. We propose experiments for two levels of inquiry. The fine level involves: 1) identifying interactions between nuclear factor(s) and DNA elements(s) shown to be necessary for in vivo induction of beta myosin heavy chain by mechanical overload, using mobility shift, DNAse I footprinting and methylation interference assays, and 2) isolating cDNA(s) encoding nuclear factor(s) involved in beta myosin heavy chain induction by mechanical overload using expression cloning. Northern analysis will determine developmental, tissue and fiber specific expression patterns of these nuclear factor(s). The gross level involves the generation and analysis of transgenic mice harboring: 1) transgenes carrying PCR site-directed mutations of DNA element(s) involved in beta myosin heavy chain induction by mechanical overload. Transgene promoter activity will be measured by assaying for chloramphenicol acetyltransferase specific activity following mechanical overload and in muscles containing various fiber types, and 2) transgenes overexpressing cDNA(s) encoding nuclear factor(s) specifically targeted to striated muscle. Northern analysis will assess the impact of overexpression of nuclear factors on striated muscle phenotype. The proposed work will identify and test the in vivo function of nuclear factor(s) and beta myosin heavy chain promoter element(s) involved in beta myosin heavy chain fiber specific expression and induction by mechanical overload. In vivo overexpression of nuclear factor(s) will identify their potential roles in regulation of other gene(s), muscle enlargement and development. These experiments are expected to identify potential protein/DNA targets for therapies aimed at providing countermeasures to derangements in muscle phenotypes and growth patterns attributable to muscle diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR041464-08
Application #
2700218
Study Section
Respiratory and Applied Physiology Study Section (RAP)
Project Start
1992-03-03
Project End
1998-09-01
Budget Start
1998-05-01
Budget End
1998-09-01
Support Year
8
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of Illinois Urbana-Champaign
Department
Physiology
Type
Schools of Arts and Sciences
DUNS #
041544081
City
Champaign
State
IL
Country
United States
Zip Code
61820
Southard, Sheryl; Kim, Ju-Ryoung; Low, SiewHui et al. (2016) Myofiber-specific TEAD1 overexpression drives satellite cell hyperplasia and counters pathological effects of dystrophin deficiency. Elife 5:
Tsika, Richard W; Ma, Lixin; Kehat, Izhak et al. (2010) TEAD-1 overexpression in the mouse heart promotes an age-dependent heart dysfunction. J Biol Chem 285:13721-35
Tsika, Richard W; Schramm, Christine; Simmer, Gretchen et al. (2008) Overexpression of TEAD-1 in transgenic mouse striated muscles produces a slower skeletal muscle contractile phenotype. J Biol Chem 283:36154-67
Ji, Juan; Tsika, Gretchen L; Rindt, Hansjorg et al. (2007) Puralpha and Purbeta collaborate with Sp3 to negatively regulate beta-myosin heavy chain gene expression during skeletal muscle inactivity. Mol Cell Biol 27:1531-43
Tsika, Gretchen; Ji, Juan; Tsika, Richard (2004) Sp3 proteins negatively regulate beta myosin heavy chain gene expression during skeletal muscle inactivity. Mol Cell Biol 24:10777-91
Karasseva, Natalia; Tsika, Gretchen; Ji, Juan et al. (2003) Transcription enhancer factor 1 binds multiple muscle MEF2 and A/T-rich elements during fast-to-slow skeletal muscle fiber type transitions. Mol Cell Biol 23:5143-64
Tsika, Richard W; McCarthy, John; Karasseva, Natalia et al. (2002) Divergence in species and regulatory role of beta -myosin heavy chain proximal promoter muscle-CAT elements. Am J Physiol Cell Physiol 283:C1761-75
Vyas, D R; McCarthy, J J; Tsika, G L et al. (2001) Multiprotein complex formation at the beta myosin heavy chain distal muscle CAT element correlates with slow muscle expression but not mechanical overload responsiveness. J Biol Chem 276:1173-84
Vyas, D R; McCarthy, J J; Tsika, R W (1999) Nuclear protein binding at the beta-myosin heavy chain A/T-rich element is enriched following increased skeletal muscle activity. J Biol Chem 274:30832-42
McCarthy, J J; Vyas, D R; Tsika, G L et al. (1999) Segregated regulatory elements direct beta-myosin heavy chain expression in response to altered muscle activity. J Biol Chem 274:14270-9

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