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Genetics, Vol 147, 1003-1016, Copyright © 1997
INVESTIGATIONS |
SRO9, a Multicopy Suppressor of the Bud Growth Defect in the Saccharomyces cerevisiae rho3-Deficient Cells, Shows Strong Genetic Interactions With Tropomyosin Genes, Suggesting Its Role in Organization of the Actin Cytoskeleton
M. Kagami, A. Toh-e and Y. Matsui
Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Hongo, Tokyo 113, Japan
RHO3 encodes a Rho-type small GTPase in the yeast Saccharomyces cerevisiae and is involved in the proper organization of the actin cytoskeleton required for bud growth. SRO9 (YCL37c) was isolated as a multicopy suppressor of a rho3{delta} mutation. An Sro9p domain required for function is similar to a domain in the La protein (an RNA-binding protein). Disruption of SRO9 did not affect vegetative growth, even with the simultaneous disruption of an SRO9 homologue, SRO99. However, sro9{delta} was synthetically lethal with a disruption of TPM1, which encodes tropomyosin; sro9{delta} tpm1{delta} cells did not distribute cortical actin patches properly and lysed. We isolated TPM2, the other gene for tropomyosin, as a multicopy suppressor of a tpm1{delta} sro9{delta} double mutant. Genetic analysis suggests that TPM2 is functionally related to TPM1 and that tropomyosin is important but not essential for cell growth. Overexpression of SRO9 suppressed the growth defect in tpm1{delta} tpm2{delta} cells, disappearance of cables of actin filaments in both rho3{delta} cells and tpm1{delta} cells, and temperature sensitivity of actin mutant cells (act1-1 cells), suggesting that Sro9p has a function that overlaps or is related to tropomyosin function. Unlike tropomyosin, Sro9p does not colocalize with actin cables but is diffusely cytoplasmic. These results suggest that Sro9p is a new cytoplasmic factor involved in the organization of actin filaments.
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