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MOLECULAR AND CELLULAR BIOLOGY, Sept. 1994, p. 6337-6349 Vol. 14, No. 9 0270-7306/94/$04.00+0

Summary: MOLECULAR AND CELLULAR BIOLOGY, Sept. 1994, p. 6337-6349 Vol. 14, No. 9
Copyright C 1994, American Society for Microbiology
Interactions between Highly Conserved U2 Small Nuclear RNA
Structures and Prp5p, Prp9p, Prpllp, and Prp2lp Proteins Are
Required To Ensure Integrity of the U2 Small Nuclear
Ribonucleoprotein in Saccharomyces cerevisiae
Sinsheimer Laboratories, University of Califomia, Santa Cruz, Santa Cruz, Califomia 95064
Received 21 April 1994/Returned for modification 6 June 1994/Accepted 15 June 1994
Binding of U2 small nuclear ribonucleoprotein (snRNP) to the pre-mRNA is an early and important step in
spliceosome assembly. We searched for evidence of cooperative function between yeast U2 small nuclear RNA
(snRNA) and several genetically identified splicing (Prp) proteins required for the first chemical step of
splicing, using the phenotype of synthetic lethality. We constructed yeast strains with pairwise combinations
of28 different U2 alleles with 10prp mutations and found lethal double-mutant combinations withprpS, -9, -11,
and -21 but not with prp3, 4, -8, or -19. Many U2 mutations in highly conserved or invariant RNA structures
show no phenotype in a wild-type PRP background but render mutantprp strains inviable, suggesting that the
conserved but dispensable U2 elements are essential for efficient cooperative function with specific Prp
proteins. Mutant U2 snRNA fails to accumulate in synthetic lethal strains, demonstrating that interaction
between U2 RNA and these four Prp proteins contributes to U2 snRNP assembly or stability. Three of the


Source: Ares Jr., Manny - Department of Molecular, Cell, and Developmental Biology, University of California at Santa Cruz


Collections: Biology and Medicine