Our long-term goal is to utilize the telokin promoter as a model system to determine the mechanisms regulating smooth muscle-specific gene expression. Unraveling these mechanisms is crucial to our understanding of smooth muscle development and of the pathology of many different vascular, pulmonary, intestinal and urogenital diseases that are associated with altered contractile protein expression and altered contractility. Experiments are proposed to test the hypothesis that modular transcription elements control gene expression in different smooth muscle tissues. Transgenic mice will be utilized to identify the minimal cell-specific telokin promoter. Important cis-acting regulatory elements identified in vitro, will be characterized by mutational analysis, of telokin promoter- beta-galactosidase transgenes. To determine if regulatory elements are important for basal expression or smooth muscle specificity we will employ a novel approach using chimeric telokin/SM22a promoters. Several transcription factors that regulate the telokin promoter activity have been identified, including SRF. HFH-l, HFH-8, CDP, TEF and HMGI. A major goal of this proposal is to determine the role played by these factors in regulating expression of telokin and other smooth muscle contractile proteins in normal and injured, phenotypically modified, smooth muscle cells. Specifically we will test the hypothesis that HFH-l and CDP repress and HFH-8 and TEF stimulate telokin gene expression. The role of architectural transcription factors HMGI(Y) in regulating telokin promoter activity and contractile protein expression will also be evaluated. SRF is known to be a key regulator of all smooth muscle genes thus far examined, although it is not known if it is required for basal expression of smooth muscle genes or if it is involved in mediating their tissue specific expression. It is likely that the tissue specific functions of SRF result from its interaction with other cell-type specific factors, hence, modified yeast-2 hybrid screens are proposed to identify novel smooth muscle restricted binding partners. Preliminary results have yielded two interesting proteins, UBC9 and Duplin, which although widely expressed, may have fundamental effects on SRF's activity through Sumo modification and interaction with beta--catenin, respectively. The function of SRFs' interaction with these proteins will be elucidated.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL058571-07
Application #
6719053
Study Section
Pathology A Study Section (PTHA)
Program Officer
Rabadan-Diehl, Cristina
Project Start
1998-04-01
Project End
2006-03-31
Budget Start
2004-04-01
Budget End
2005-03-31
Support Year
7
Fiscal Year
2004
Total Cost
$298,000
Indirect Cost
Name
Indiana University-Purdue University at Indianapolis
Department
Physiology
Type
Schools of Medicine
DUNS #
603007902
City
Indianapolis
State
IN
Country
United States
Zip Code
46202
Zhou, Jiliang; Zhang, Min; Fang, Hong et al. (2009) The SWI/SNF chromatin remodeling complex regulates myocardin-induced smooth muscle-specific gene expression. Arterioscler Thromb Vasc Biol 29:921-8
Herring, B Paul; Zhou, Jiliang (2007) mCAT got youR TEF? Circ Res 101:856-8
Zhang, Min; Fang, Hong; Zhou, Jiliang et al. (2007) A novel role of Brg1 in the regulation of SRF/MRTFA-dependent smooth muscle-specific gene expression. J Biol Chem 282:25708-16
Touw, Ketrija; Hoggatt, April M; Simon, Gina et al. (2007) Hprt-targeted transgenes provide new insights into smooth muscle-restricted promoter activity. Am J Physiol Cell Physiol 292:C1024-32
Yin, Feng; Hoggatt, April M; Zhou, Jiliang et al. (2006) 130-kDa smooth muscle myosin light chain kinase is transcribed from a CArG-dependent, internal promoter within the mouse mylk gene. Am J Physiol Cell Physiol 290:C1599-609
Herring, B Paul; El-Mounayri, Omar; Gallagher, Patricia J et al. (2006) Regulation of myosin light chain kinase and telokin expression in smooth muscle tissues. Am J Physiol Cell Physiol 291:C817-27
El-Mounayri, Omar; Triplett, Jason W; Yates, Charles W et al. (2005) Regulation of smooth muscle-specific gene expression by homeodomain proteins, Hoxa10 and Hoxb8. J Biol Chem 280:25854-63
Zhou, Jiliang; Herring, B Paul (2005) Mechanisms responsible for the promoter-specific effects of myocardin. J Biol Chem 280:10861-9
Yin, Feng; Herring, B Paul (2005) GATA-6 can act as a positive or negative regulator of smooth muscle-specific gene expression. J Biol Chem 280:4745-52
Triplett, Jason W; Herring, B Paul; Pavalko, Fredrick M (2005) Adenoviral transgene expression enhanced by cotreatment with etoposide in cultured cells. Biotechniques 39:826, 828, 830, passim

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