Identification of Novel Genes Required for Yeast Pre-mRNA Splicing by Means of Cold-Sensitive Mutations

INVESTIGATIONS

Identification of Novel Genes Required for Yeast Pre-mRNA Splicing by Means of Cold-Sensitive Mutations

S. M. Noble and C. Guthrie
Department of Biochemistry and Biophysics, University of California, San Francisco, California, 94143-0448

Genetic approaches in Saccharomyces cerevisiae have identified 38 genes required for efficient RNA splicing. The majority have been found by screening (high) temperature-sensitive (ts) mutants for those defective in splicing, an approach limited by the presence of ts hotspots and by the fact that many essential genes rarely mutate to the ts phenotype. To identify novel genes, we screened a collection of 340 cold-sensitive (cs) mutants for those that exhibited diminished splicing of several pre-mRNAs. We isolated 12 mutants in nine complementation groups. Four of these affected known genes (PRP8, PRP16, PRP22, PRP28), three of which encode RNA helicase homologues. Five genes are novel (BRR1, BRR2, BRR3, BRR4, BRR5; Bad Response to Refrigeration); mutations in these genes inhibited splicing before the first chemical step of the reaction. Analysis of BRR2 revealed it to encode an essential member of a new class of RNA helicase-like proteins that includes the yeast antiviral protein Ski2. These data validate the use of cs mutants in genetic screens and raise the possibility that RNA helicase family members are particularly prone to mutation to cold sensitivity.

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				J. Zhao, M. Kessler, S. Helmling, J. P. O'Connor, and C. Moore Pta1, a Component of Yeast CF II, Is Required for Both Cleavage and Poly(A) Addition of mRNA Precursor Mol. Cell. Biol., November 1, 1999; 19(11): 7733 - 7740. [Abstract] [Full Text] [PDF]

				D.-H. Kim, G. Edwalds-Gilbert, C. Ren, and R.-J. Lin A Mutation in a Methionine tRNA Gene Suppresses the prp2-1 Ts Mutation and Causes a Pre-mRNA Splicing Defect in Saccharomyces cerevisiae Genetics, November 1, 1999; 153(3): 1105 - 1115. [Abstract] [Full Text]

				S. W. Stevens and J. Abelson Purification of the yeast U4/U6·U5 small nuclear ribonucleoprotein particle and identification of its proteins PNAS, June 22, 1999; 96(13): 7226 - 7231. [Abstract] [Full Text] [PDF]

				T. Achsel, K. Ahrens, H. Brahms, S. Teigelkamp, and R. Lührmann The Human U5-220kD Protein (hPrp8) Forms a Stable RNA-Free Complex with Several U5-Specific Proteins, Including an RNA Unwindase, a Homologue of Ribosomal Elongation Factor EF-2, and a Novel WD-40 Protein Mol. Cell. Biol., November 1, 1998; 18(11): 6756 - 6766. [Abstract] [Full Text]

				F. Stutz and M. Rosbash Nuclear RNA export Genes & Dev., November 1, 1998; 12(21): 3303 - 3319. [Full Text]

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				P. L. Raghunathan and C. Guthrie A Spliceosomal Recycling Factor That Reanneals U4 and U6 Small Nuclear Ribonucleoprotein Particles Science, February 6, 1998; 279(5352): 857 - 860. [Abstract] [Full Text]

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