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PMID: 1409649 Published · ppublish English Comparative Study Journal Article

Dominant negative selection of heterologous genes: isolation of Candida albicans genes that interfere with Saccharomyces cerevisiae mating factor-induced cell cycle arrest.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 89 ·No. 20 ·1992-10-15 ·Pages 9410-4

Whiteway M, Dignard D, Thomas DY

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.

MeSH Terms
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 Factor Mitogen-Activated Protein Kinase 3 Mitogen-Activated Protein Kinases Molecular Sequence Data Peptides/genetics Protein-Tyrosine Kinases/genetics Proteins/genetics Restriction Mapping Saccharomyces cerevisiae/genetics Sequence Alignment Species Specificity Transcription Factors/genetics Zinc Fingers
Chemicals
Cyclins DNA, Fungal Fungal Proteins Peptides Proteins Transcription Factors CEK1 protein, Candida albicans CZF1 protein, Candida albicans CLN1 protein, Candida albicans CRL1 protein, Candida albicans Mating Factor Protein-Tyrosine Kinases Calcium-Calmodulin-Dependent Protein Kinases Mitogen-Activated Protein Kinase 3 Mitogen-Activated Protein Kinases GTP-Binding Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Whiteway M
Eukaryotic Genetics Group, National Research Council of Canada, Montréal, Québec.
Dignard D
Thomas D Y
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.89.20.9410
Published
1992-10-15
Pages
9410-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC50141
Subset
IM
Databases
GENBANK
M76585, M76586, M76587, M83991
External Links
PubMed source DOI: 10.1073/pnas.89.20.9410 PMC Full Text View journal details
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