Cooperation of two actin-binding proteins, cofilin and Aip1, in Saccharomyces cerevisiae.
BACKGROUND: Cofilin is a low-molecular weight actin-modulating protein, and is structurally and functionally conserved among eukaryotes. Cofilin is encoded by COF1 in Saccharomyces cerevisiae, and is essential for cell viability. Cofilin binds to and severs actin filaments in vitro, and also enhances their depolymerization. A partner protein that cooperates with ... cofilin in vivo has not been identified. RESULTS: When COF1 was over-expressed in yeast cells under the GAL1 promoter in a medium containing galactose as a sole carbon source, the cells did not survive. These results indicate that cells can grow only when the expression of cofilin is appropriately regulated. Several temperature sensitive (ts-) mutants were independently created by the random mutagenesis of COF1 with hydroxylamine. Mutated amino acids in ts-mutants were mapped in the sequences that were presumed to be involved in actin binding. A gene on a multicopy plasmid which suppresses the ts-phenotype of cof1-101, a typical ts-cofilin mutant, was isolated. The suppressor gene, SCF1, was found to be identical to AIP1, a gene encoding an actin-interacting protein. Although SCF1/AIP1 is not essential for cell viability, a combination of cof1-101 and Deltascf1/aip1 is synthetic lethal. Immunofluorescence staining of a wild-type strain using anti-Aip1 antibodies revealed that Aip1 was distributed in cortical actin patches where cofilin was also co-localized. Thick and long fibres stained with anti-cofilin antibody were detected in Deltascf1/aip1 cells, but not in SCF1/AIP1 cells. CONCLUSIONS: These results suggest the cooperative modulation of the actin cytoskeleton by cofilin and Aip1.
Mesh Terms:
Actin Depolymerizing Factors, Actins, Amino Acid Sequence, Base Sequence, Cell Cycle Proteins, Cell Division, Electrophoresis, Polyacrylamide Gel, Fluorescent Antibody Technique, Fungal Proteins, Galactose, Genotype, Glucose, Heat-Shock Proteins, Microfilament Proteins, Models, Genetic, Molecular Sequence Data, Mutagenesis, Phenotype, Recombinant Proteins, Saccharomyces cerevisiae, Schizosaccharomyces pombe Proteins, Sequence Homology, Nucleic Acid, Suppression, Genetic, Temperature, Time Factors
Actin Depolymerizing Factors, Actins, Amino Acid Sequence, Base Sequence, Cell Cycle Proteins, Cell Division, Electrophoresis, Polyacrylamide Gel, Fluorescent Antibody Technique, Fungal Proteins, Galactose, Genotype, Glucose, Heat-Shock Proteins, Microfilament Proteins, Models, Genetic, Molecular Sequence Data, Mutagenesis, Phenotype, Recombinant Proteins, Saccharomyces cerevisiae, Schizosaccharomyces pombe Proteins, Sequence Homology, Nucleic Acid, Suppression, Genetic, Temperature, Time Factors
Genes Cells
Date: Jan. 01, 1999
PubMed ID: 10231390
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