The overall objective of the research proposed in this application is to elucidate molecular mechanisms used to regulate pre-mRNA splicing during development of higher metazoan organism using a combination of molecular biology and biochemistry.
The specific aims are as follows: 1. Refine biochemical reconstitution assays to study the somatic repression of P element IVS3 pre-mRNA splicing in vitro. 2. Identify and characterize the 65 kD and 40 kD RNA-binding proteins that are part of the P element IVS3 5' exon inhibitory complex. 3. Characterize the biochemical properties of the recombinant PSI (97 kD) and hrp48 proteins. 4. Analyze the expression patterns of PSI and isolate P element insertion mutations in the PSI gene; characterize mutations in the hrp48 gene.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD028063-08
Application #
2403237
Study Section
Special Emphasis Panel (ZRG2-CTY (02))
Project Start
1991-07-01
Project End
2000-07-31
Budget Start
1997-08-01
Budget End
1998-07-31
Support Year
8
Fiscal Year
1997
Total Cost
Indirect Cost
Name
University of California Berkeley
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
094878337
City
Berkeley
State
CA
Country
United States
Zip Code
94704
Labourier, E; Rio, D C (2001) Purification of Drosophila snRNPs and characterization of two populations of functional U1 particles. RNA 7:457-70
Crowder, S M; Kanaar, R; Rio, D C et al. (1999) Absence of interdomain contacts in the crystal structure of the RNA recognition motifs of Sex-lethal. Proc Natl Acad Sci U S A 96:4892-7
Rudner, D Z; Kanaar, R; Breger, K S et al. (1998) Interaction between subunits of heterodimeric splicing factor U2AF is essential in vivo. Mol Cell Biol 18:1765-73
Rudner, D Z; Breger, K S; Rio, D C (1998) Molecular genetic analysis of the heterodimeric splicing factor U2AF: the RS domain on either the large or small Drosophila subunit is dispensable in vivo. Genes Dev 12:1010-21
Lee, A L; Volkman, B F; Robertson, S A et al. (1997) Chemical shift mapping of the RNA-binding interface of the multiple-RBD protein sex-lethal. Biochemistry 36:14306-17
Adams, M D; Tarng, R S; Rio, D C (1997) The alternative splicing factor PSI regulates P-element third intron splicing in vivo. Genes Dev 11:129-38
Hammond, L E; Rudner, D Z; Kanaar, R et al. (1997) Mutations in the hrp48 gene, which encodes a Drosophila heterogeneous nuclear ribonucleoprotein particle protein, cause lethality and developmental defects and affect P-element third-intron splicing in vivo. Mol Cell Biol 17:7260-7
Kanaar, R; Lee, A L; Rudner, D Z et al. (1995) Interaction of the sex-lethal RNA binding domains with RNA. EMBO J 14:4530-9
Siebel, C W; Admon, A; Rio, D C (1995) Soma-specific expression and cloning of PSI, a negative regulator of P element pre-mRNA splicing. Genes Dev 9:269-83
Lee, A L; Kanaar, R; Rio, D C et al. (1994) Resonance assignments and solution structure of the second RNA-binding domain of sex-lethal determined by multidimensional heteronuclear magnetic resonance. Biochemistry 33:13775-86

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