Hsp90 protein in fission yeast Swo1p and UCS protein Rng3p facilitate myosin II assembly and function.
The F-actin-based molecular motor myosin II is involved in a variety of cellular processes such as muscle contraction, cell motility, and cytokinesis. In recent years, a family of myosin II-specific cochaperones of the UCS family has been identified from work with yeasts, fungi, worms, and humans. Biochemical analyses have shown ... that a complex of Hsp90 and the Caenorhabditis elegans UCS domain protein UNC-45 prevent myosin head aggregation, thereby allowing it to assume a proper structure. Here we demonstrate that a temperature-sensitive mutant of the fission yeast Hsp90 (Swo1p), swo1-w1, is defective in actomyosin ring assembly at the restrictive temperature. Two alleles of swo1, swo1-w1 and swo1-26, showed synthetic lethality with a specific mutant allele of the fission yeast type II myosin head, myo2-E1, but not with two other mutant alleles of myo2 or with mutations affecting 14 other genes important for cytokinesis. swo1-w1 also showed a strong genetic interaction with rng3-65, a gene encoding a mutation in the fission yeast UCS domain protein Rng3p, which has previously been shown to be important for myosin II assembly. A similar deleterious effect was found when myo2-E1, swo1-w1, and rng3-65 were pharmacologically treated with geldanamycin to partially inhibit Hsp90 function. Interestingly, Swo1p-green fluorescent protein is detected at the improperly assembled actomyosin rings in myo2-E1 but not in a wild-type strain. Yeast two-hybrid and coimmunoprecipitation analyses verified interactions between Rng3p and the myosin head domain as well as interactions between Rng3p and Swo1p. Our analyses of Myo2p, Swo1p, and the UCS domain protein Rng3p establish that Swo1p and Rng3p collaborate in vivo to modulate myosin II function.
Mesh Terms:
Alleles, Animals, Benzoquinones, Cytoskeletal Proteins, Cytoskeleton, Enzyme Inhibitors, HSP90 Heat-Shock Proteins, Humans, Lactams, Macrocyclic, Myosin Heavy Chains, Myosin Type II, Protein Binding, Protein Structure, Tertiary, Quinones, Recombinant Fusion Proteins, Schizosaccharomyces, Schizosaccharomyces pombe Proteins, Temperature, Two-Hybrid System Techniques
Alleles, Animals, Benzoquinones, Cytoskeletal Proteins, Cytoskeleton, Enzyme Inhibitors, HSP90 Heat-Shock Proteins, Humans, Lactams, Macrocyclic, Myosin Heavy Chains, Myosin Type II, Protein Binding, Protein Structure, Tertiary, Quinones, Recombinant Fusion Proteins, Schizosaccharomyces, Schizosaccharomyces pombe Proteins, Temperature, Two-Hybrid System Techniques
Eukaryotic Cell
Date: Mar. 01, 2005
PubMed ID: 15755919
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