| Title: | Dominant negative selection of heterologous genes: isolation of Candida albicans genes that interfere with Saccharomyces cerevisiae mating factor-induced cell cycle arrest |
| Author(s): | Whiteway M; Dignard D; Thomas DY; |
| Address: | "Eukaryotic Genetics Group, National Research Council of Canada, Montreal, Quebec" |
| ISSN/ISBN: | 0027-8424 (Print) 1091-6490 (Electronic) 0027-8424 (Linking) |
| Abstract: | "We have used a genomic library of Candida albicans to transform Saccharomyces cerevisiae and screened for genes that act similarly to dominant negative mutations by interfering with pheromone-mediated cell cycle arrest. Six different plasmids were identified from 2000 transformants; four have been sequenced. One gene (CZF1) encodes a protein with structural motifs characteristic of a transcription factor. A second gene (CCN1) encodes a cyclin homologue, a third (CRL1) encodes a protein with sequence similarity to GTP-binding proteins of the RHO family, and a fourth (CEK1) encodes a putative kinase of the ERK family. Since CEK1 confers a phenotype similar to that of the structurally related S. cerevisiae gene KSS1 but cannot complement a KSS1 defect, it is evident that dominant negative selection can identify proteins that complementation screens would miss. Because dominant negative mutations exert their influence even in wild-type strain backgrounds, this approach should be a general method for the analysis of complex cellular processes in organisms not amenable to direct genetic analysis" |
| Keywords: | "Amino Acid Sequence Calcium-Calmodulin-Dependent Protein Kinases Candida albicans/*genetics *Cell Cycle Consensus Sequence Cyclins/genetics DNA, Fungal/genetics Fungal Proteins/*genetics GTP-Binding Proteins/genetics Genes, Dominant *Genes, Fungal Mating;" |
| Notes: | "MedlineWhiteway, M Dignard, D Thomas, D Y eng Comparative Study 1992/10/15 Proc Natl Acad Sci U S A. 1992 Oct 15; 89(20):9410-4. doi: 10.1073/pnas.89.20.9410" |
