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Nucleic Acids Research, 2002, Vol. 30, No. 22 4993-5003
© 2002 Oxford University Press

Thermoconditional modulation of the pleiotropic sensitivity phenotype by the Saccharomyces cerevisiae PRP19 mutant allele pso4-1

L. F. Revers, J. M. Cardone, D. Bonatto, J. Saffi, M. Grey1, H. Feldmann2, M. Brendel1 and J. A. P. Henriques*

Depto. de Biofísica/Centro de Biotecnologia–IB-UFRGS, Avenida Bento Gonçalves, 9500, Prédio 43421, Campus do Vale, 91501-907 Porto Alegre, RS, Brazil, 1 Institut für Mikrobiologie, Abteilung Biologie f. Mediziner, Johann Wolfgang Goethe-Universität, Theodor-Stern-Kai 7, Haus 75, 60590 Frankfurt/Main, Germany and 2 Institut für Biochemie der Ludwig-Maximilians Universität, München, Feodor-Lynen-Strasse 25, 81377 München, Germany

*To whom correspondence should be addressed. Tel: +55 51 3316 6069; Fax: +55 51 3316 6084; Email: pegas{at}dna.cbiot.ufrgs.br

The conditionally-lethal pso4-1 mutant allele of the spliceosomal-associated PRP19 gene allowed us to study this gene’s influence on pre-mRNA processing, DNA repair and sporulation. Phenotypes related to intron-containing genes were correlated to temperature. Splicing reporter systems and RT–PCR showed splicing efficiency in pso4-1 to be inversely correlated to growth temperature. A single amino acid substitution, replacing leucine with serine, was identified within the N-terminal region of the pso4-1 allele and was shown to affect the interacting properties of Pso4-1p. Amongst 24 interacting clones isolated in a two-hybrid screening, seven could be identified as parts of the RAD2, RLF2 and DBR1 genes. RAD2 encodes an endonuclease indispensable for nucleotide excision repair (NER), RLF2 encodes the major subunit of the chromatin assembly factor I, whose deletion results in sensitivity to UVC radiation, while DBR1 encodes the lariat RNA splicing debranching enzyme, which degrades intron lariat structures during splicing. Characterization of mutagen-sensitive phenotypes of rad2{Delta}, rlf2{Delta} and pso4-1 single and double mutant strains showed enhanced sensitivity for the rad2{Delta} pso4-1 and rlf2{Delta} pso4-1 double mutants, suggesting a functional interference of these proteins in DNA repair processes in Saccharomyces cerevisiae.


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