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PMID:8294515
| Citation |
Brockerhoff, SE, Stevens, RC and Davis, TN (1994) The unconventional myosin, Myo2p, is a calmodulin target at sites of cell growth in Saccharomyces cerevisiae. J. Cell Biol. 124:315-23 |
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| Abstract |
Myo2p is an unconventional myosin required for polarized growth in Saccharomyces cerevisiae. Four lines of evidence suggest that (a) Myo2p is a target of calmodulin at sites of cell growth, and (b) the interaction between Myo2p and calmodulin is Ca2+ independent. First, as assessed by indirect immunofluorescence, the distributions of Myo2p and calmodulin are nearly indistinguishable throughout the cell cycle. Second, a genetic analysis indicates that mutations in CMD1 show allele-specific synthetic lethality with the myo2-66 conditional mutation. Mutations that inactivate the Ca(2+)-binding sites of calmodulin have little or no effect on strains carrying myo2-66, whereas an allele with a mutation outside the Ca(2+)-binding sites dramatically increases the severity of the phenotype conferred by myo2-66. Third, Myo2p coimmunoprecipitates with calmodulin in the presence of Ca2+ or EGTA. Finally, we used a modified gel overlay assay to demonstrate direct interaction between calmodulin and fusion proteins containing portions of Myo2p. Calmodulin binds specifically to the region of Myo2p containing six tandem repeats of a motif called an IQ site. Binding occurs in either Ca2+ or EGTA, and only two sites are required to observe binding. |
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| Keywords |
Alleles; Base Sequence; Calcium/metabolism; Calmodulin/genetics; Calmodulin/metabolism; Carrier Proteins/genetics; Carrier Proteins/metabolism; Cell Division; Cytoskeletal Proteins; Egtazic Acid/pharmacology; Fluorescent Antibody Technique; Fungal Proteins/genetics; Fungal Proteins/metabolism; Molecular Sequence Data; Mutation; Myosin Heavy Chains; Myosin Type II; Myosin Type V; Recombinant Fusion Proteins/metabolism; Saccharomyces cerevisiae/cytology; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae/growth & development; Saccharomyces cerevisiae/metabolism; Saccharomyces cerevisiae Proteins; Schizosaccharomyces pombe Proteins |
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Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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