Functional connection between the Clb5 cyclin, the protein kinase C pathway and the Swi4 transcription factor in Saccharomyces cerevisiae.
The rsf12 mutation was isolated in a synthetic lethal screen for genes functionally interacting with Swi4. RSF12 is CLB5. The clb5 swi4 mutant cells arrest at G(2)/M due to the activation of the DNA-damage checkpoint. Defects in DNA integrity was confirmed by the increased rates of chromosome loss and mitotic recombination. Other results suggest the presence of additional defects related to morphogenesis. Interestingly, genes of the PKC pathway rescue the growth defect of clb5 swi4, and pkc1 and slt2 mutations are synthetic lethal with clb5, pointing to a connection between Clb5, the PKC pathway, and Swi4. Different observations suggest that like Clb5, the PKC pathway and Swi4 are involved in the control of DNA integrity: there is a synthetic interaction between pkc1 and slt2 with rad9; the pkc1, slt2, and swi4 mutants are hypersensitive to hydroxyurea; and the Slt2 kinase is activated by hydroxyurea. Reciprocally, we found that clb5 mutant is hypersensitive to SDS, CFW, latrunculin B, or zymolyase, which suggests that, like the PKC pathway and Swi4, Clb5 is related to cell integrity. In summary, we report numerous genetic interactions and phenotypic descriptions supporting a close functional relationship between the Clb5 cyclin, the PKC pathway, and the Swi4 transcription factor.
Queralt
Ethel
E
Departament de Bioquímica i Biologia Molecular, Universitat de València, C/Dr. Moliner 50, E-46100 Burjassot, Valencia, Spain.
Igual
J Carlos
JC
D015153:Blotting, Western; D002453:Cell Cycle / Q000235:genetics* / Q000502:physiology; D015825:Chromosomes, Fungal / Q000235:genetics*; D019926:Cyclin B / Q000235:genetics / Q000378:metabolism*; D004271:DNA, Fungal / Q000737:chemistry*; D004268:DNA-Binding Proteins; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D006918:Hydroxyurea; D047468:Immunoprecipitation; D009154:Mutation / Q000235:genetics; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D011995:Recombination, Genetic / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
C076558:CLB5 protein, S cerevisiae; D019926:Cyclin B; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; C062214:SWI4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D011493:Protein Kinase C; D006918:Hydroxyurea