Nucleic Acids Research, 2003, Vol. 31, No. 9 2333-2343
© 2003 Oxford University Press
Dependence of pre-mRNA introns on PRP17, a non-essential splicing factor: implications for efficient progression through cell cycle transitions
Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560012, India and 1 Institute of Molecular and Cell Biology, 30 Medical Drive, Singapore 117609, Singapore
*To whom correspondence should be addressed. Tel: +11 91 80 3600168; Fax: +11 91 80 3602697; Email: uvr{at}mcbl.iisc.ernet.in
Saccharomyces cerevisiae PRP17 (CDC40) encodes a second-step pre-mRNA splicing factor with a role in cell division. The functions of Prp17 in specific cell cycle transitions were examined using temperature-sensitive alleles in arrest/release experiments. We find that G1/S and G2/M transitions depend on Prp17. G1-synchronized prp17::LEU2 cells arrest at non-permissive temperatures as unbudded haploid cells with low levels of CLN1, CLB5 and RNR1 transcripts. This indicates a Prp17 execution point at or prior to Start. Reduced levels of 
-tubulin protein, a mitotic spindle component, underlie the benomyl sensitivity of prp17 mutants and possibly their G2/M arrest. Splicing of TUB1 and TUB3 transcripts, which encode -tubulin, was analyzed in prp17 and other second-step factor mutants. TUB1 splicing is inefficient in prp17, prp16 and prp22, and marginally affected in prp18, slu7-1 and psf1-1. TUB3 splicing is similarly affected. In vitro splicing with TUB3 pre-mRNA demonstrates a compromised second step in prp17::LEU2 extracts, implicating a direct role for Prp17 in its efficient splicing. Genomic replacement of an intronless TUB1 gene relieves the benomyl sensitivity of prp17 mutants; however, they remain temperature sensitive, implying multiple limiting factors for mitosis. The data suggest that integration of splicing with the cell cycle is important for G1/S and G2/M transitions.
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