Skeletal myogenesis is characterized by the coordinated expression of a large number of genes. In skeletal muscle diseases and muscle tumors, many of these genes have an altered level of expression and emerging genomic technologies have been used to identify patterns of gene expression associated with normal differentiation and disease. Yet, the molecular basis for coordinately regulating subprograms of gene expression remains largely unknown. Our broad and long-term objective is to identify the molecular basis for the orchestrated regulation of subprograms of gene expression. Myogenesis provides an excellent model system, as it is known that the entire myogenic program can be initiated by a single transcription factor, MyoD. We have shown that MyoD directly patterns discrete subprograms of gene expression through a combination of regulated binding and promoter-specific activity. We will use our model of MyoD-induced myogenesis to test the broad hypothesis that complex patterns of gene expression are established by the regulated activity of a single transcription factor, rather than by multiple independent pathways.
Our Specific Aims will (Aim 1) use a cell culture system of MyoD-mediated myogenesis to: (a) determine the molecular regulation of promoter-specific MyoD binding; (b) identify the promoter-specific role of individual activation domains of MyoD; and (c) determine the functionally relevant isoform of p38 and its mechanism of regulating MyoD;
(Aim 2) use our in vitro system of MyoD-mediated transcription on chromatinized templates to directly test the roles of specific factors in sequential steps of transcriptional activation;
and (Aim 3) use array analysis and the mouse and human genomic sequences to identify promoter motifs associated with specific subprograms of MyoD-mediated gene expression. The significance of this proposal is that we will reveal how a complex mixture of regulatory factors are integrated by a single transcription factor to orchestrate a coordinated pattern of gene expression. The health relatedness of this proposal is that the identification of the molecular basis of patterned gene expression will provide a fundamental framework for the study of gene expression in human development and disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
2R01AR045113-06
Application #
6574081
Study Section
Special Emphasis Panel (ZRG1-SMB (01))
Program Officer
Nuckolls, Glen H
Project Start
1998-05-01
Project End
2008-01-31
Budget Start
2003-02-01
Budget End
2004-01-31
Support Year
6
Fiscal Year
2003
Total Cost
$402,021
Indirect Cost
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
078200995
City
Seattle
State
WA
Country
United States
Zip Code
98109
Conerly, Melissa L; Yao, Zizhen; Zhong, Jun Wen et al. (2016) Distinct Activities of Myf5 and MyoD Indicate Separate Roles in Skeletal Muscle Lineage Specification and Differentiation. Dev Cell 36:375-85
Fong, Abraham P; Yao, Zizhen; Zhong, Jun Wen et al. (2015) Conversion of MyoD to a neurogenic factor: binding site specificity determines lineage. Cell Rep 10:1937-46
Yang, Z J P; Broz, D Kenzelmann; Noderer, W L et al. (2015) p53 suppresses muscle differentiation at the myogenin step in response to genotoxic stress. Cell Death Differ 22:560-73
Yao, Zizhen; Macquarrie, Kyle L; Fong, Abraham P et al. (2014) Discriminative motif analysis of high-throughput dataset. Bioinformatics 30:775-83
Yao, Zizhen; Farr 3rd, Gist H; Tapscott, Stephen J et al. (2013) Pbx and Prdm1a transcription factors differentially regulate subsets of the fast skeletal muscle program in zebrafish. Biol Open 2:546-55
Diede, Scott J; Yao, Zizhen; Keyes, C Chip et al. (2013) Fundamental differences in promoter CpG island DNA hypermethylation between human cancer and genetically engineered mouse models of cancer. Epigenetics 8:1254-60
Dey, Joyoti; Dubuc, Adrian M; Pedro, Kyle D et al. (2013) MyoD is a tumor suppressor gene in medulloblastoma. Cancer Res 73:6828-37
Fong, Abraham P; Tapscott, Stephen J (2013) Skeletal muscle programming and re-programming. Curr Opin Genet Dev 23:568-73
MacQuarrie, Kyle L; Yao, Zizhen; Fong, Abraham P et al. (2013) Comparison of genome-wide binding of MyoD in normal human myogenic cells and rhabdomyosarcomas identifies regional and local suppression of promyogenic transcription factors. Mol Cell Biol 33:773-84
Yao, Zizhen; Fong, Abraham P; Cao, Yi et al. (2013) Comparison of endogenous and overexpressed MyoD shows enhanced binding of physiologically bound sites. Skelet Muscle 3:8

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