The objectives of the experiments outlined in this proposal are to determine the mechanisms and controls regulating developmental gene expression. We have determined the structure and expression of the muscle tropomyosin I (mTm I) gene from Drosophila melanogaster. The gene consists of 5 exons and 4 introns. During myogenesis, the gene is developmentally regulated and undergoes alternative splicing in different muscle of the fly. A set of experiments are proposed to study promoter function of the mTm I gene by P element mediated transformation. The expression of the normal and in vitro mutated gene will be determined in transformed flies in order to define the physical limits of the gene and DNA sequences involved in expression. Promoter function will also be studied in an in vitro cell-free transcription system to identify and characterize trans-acting regulatory factors. A second series of experiments will focus on the regulation and mechanisms that control alternative splicing. These experiments will involve an analysis of intron, exon and DNA sequence requirements necessary for proper splicing in different muscle. Normal and in vitro mutated gene transcription and RNA processing will be studied in transformed flies and cell-free extracts. The rationale for these experiments parallels the proposed studies on promoter function. A third objective of this proposal is to finish our analysis of the cytoplasmic-muscle tropomyosin II gene complex. These experiments will involve cDNA isolation, S1 nuclease mapping and DNA sequencing. Finally, the identity of a putative troponin C clone will be confirmed and the gene structure determined.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM027611-10
Application #
3274819
Study Section
Genetics Study Section (GEN)
Project Start
1979-08-06
Project End
1990-07-31
Budget Start
1988-08-01
Budget End
1989-07-31
Support Year
10
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Illinois at Chicago
Department
Type
Overall Medical
DUNS #
121911077
City
Chicago
State
IL
Country
United States
Zip Code
60612
Reddy, Karen L; Rovani, Margritte K; Wohlwill, Arthur et al. (2006) The Drosophila Par domain protein I gene, Pdp1, is a regulator of larval growth, mitosis and endoreplication. Dev Biol 289:100-14
Lin, M H; Nguyen, H T; Dybala, C et al. (1996) Myocyte-specific enhancer factor 2 acts cooperatively with a muscle activator region to regulate Drosophila tropomyosin gene muscle expression. Proc Natl Acad Sci U S A 93:4623-8
Hales, K H; Meredith, J E; Storti, R V (1994) Transcriptional and post-transcriptional regulation of maternal and zygotic cytoskeletal tropomyosin mRNA during Drosophila development correlates with specific morphogenic events. Dev Biol 165:639-53
Lichter, J B; Storti, R V (1991) In vitro transcription analysis of the Drosophila tropomyosin and other muscle genes. Biochim Biophys Acta 1088:419-24
Tansey, T; Schultz, J R; Miller, R C et al. (1991) Small differences in Drosophila tropomyosin expression have significant effects on muscle function. Mol Cell Biol 11:6337-42
Schultz, J R; Tansey, T; Gremke, L et al. (1991) A muscle-specific intron enhancer required for rescue of indirect flight muscle and jump muscle function regulates Drosophila tropomyosin I gene expression. Mol Cell Biol 11:1901-11
Hanke, P D; Storti, R V (1988) The Drosophila melanogaster tropomyosin II gene produces multiple proteins by use of alternative tissue-specific promoters and alternative splicing. Mol Cell Biol 8:3591-602
Hanke, P D; Lepinske, H M; Storti, R V (1987) Characterization of a Drosophila cDNA clone that encodes a 252-amino acid non-muscle tropomyosin isoform. J Biol Chem 262:17370-3
Hanke, P D; Storti, R V (1986) Nucleotide sequence of a cDNA clone encoding a Drosophila muscle tropomyosin II isoform. Gene 45:211-4
Basi, G S; Storti, R V (1986) Structure and DNA sequence of the tropomyosin I gene from Drosophila melanogaster. J Biol Chem 261:817-27

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