Abstract

The objective of these studies is to understand the early events in the regulation of alternative pre-mRNA splicing, particularly as it relates to the splicing of GH pre-mRNA.
The specific aims to be addressed are: 1. The molecular details of the initial enhancer recognition event will be determined, including the precise nucleotide sequence of the bGH exonic enhancer, and the role of important functional domains of SF2/ASF (RNA binding and Arg/Ser domains) will be defined. 2. The mechanism whereby the exon specific enhancer (ESE)-splicing factor complex proceeds to activate suboptimal splice sites will be defined by determining the additional splicing components that are recruited to the enhancer complex, the contribution of protein-protein interactions to this recruitment, and the role of the enhancer in bringing distant sites into the complex. 3. The GH model will be used to determine the role of changing splicing factor concentrations in developmentally-regulated alternative patterns of splicing.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK032770-16
Application #
2749437
Study Section
Biochemical Endocrinology Study Section (BCE)
Program Officer
Sato, Sheryl M
Project Start
1983-08-01
Project End
2001-07-31
Budget Start
1998-08-25
Budget End
1999-07-31
Support Year
16
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
Case Western Reserve University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106

  Publications

Li, X; Shambaugh, M E; Rottman, F M et al. (2000) SR proteins Asf/SF2 and 9G8 interact to activate enhancer-dependent intron D splicing of bovine growth hormone pre-mRNA in vitro. RNA 6:1847-58
Dirksen, W P; Li, X; Mayeda, A et al. (2000) Mapping the SF2/ASF binding sites in the bovine growth hormone exonic splicing enhancer. J Biol Chem 275:29170-7
Stallings-Mann, M L; Ludwiczak, R L; Klinger, K W et al. (1996) Alternative splicing of exon 3 of the human growth hormone receptor is the result of an unusual genetic polymorphism. Proc Natl Acad Sci U S A 93:12394-9
Dirksen, W P; Sun, Q; Rottman, F M (1995) Multiple splicing signals control alternative intron retention of bovine growth hormone pre-mRNA. J Biol Chem 270:5346-52
Dirksen, W P; Hampson, R K; Sun, Q et al. (1994) A purine-rich exon sequence enhances alternative splicing of bovine growth hormone pre-mRNA. J Biol Chem 269:6431-6
Sun, Q; Hampson, R K; Rottman, F M (1993) In vitro analysis of bovine growth hormone pre-mRNA alternative splicing. Involvement of exon sequences and trans-acting factor(s). J Biol Chem 268:15659-66
Sun, Q; Mayeda, A; Hampson, R K et al. (1993) General splicing factor SF2/ASF promotes alternative splicing by binding to an exonic splicing enhancer. Genes Dev 7:2598-608
Salata, R A; Malhotra, I J; Hampson, R K et al. (1992) Application of an immune-tolerizing procedure to generate monoclonal antibodies specific to an alternate protein isoform of bovine growth hormone. Anal Biochem 207:142-9
Goodwin, E C; Rottman, F M (1992) The 3'-flanking sequence of the bovine growth hormone gene contains novel elements required for efficient and accurate polyadenylation. J Biol Chem 267:16330-4
Helms, S R; Rottman, F M (1990) Characterization of an inducible promoter system to investigate decay of stable mRNA molecules. Nucleic Acids Res 18:255-9

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