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Eukaryotic Cell, March 2006, p. 435-446, Vol. 5, No. 3
1535-9778/06/$08.00+0     doi:10.1128/EC.5.3.435-446.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Fission Yeast Rho5p GTPase Is a Functional Paralogue of Rho1p That Plays a Role in Survival of Spores and Stationary-Phase Cells

Instituto Microbiología-Bioquímica, Departamento de Microbiología-Genética, Consejo Superior de Investigaciones Científicas/Universidad de Salamanca, 37007 Salamanca, Spain

Received 5 July 2005/ Accepted 16 December 2005

The Rho GTPase family and their effectors are key regulators involved in many eukaryotic cell functions related to actin organization and polarity establishment. Schizosaccharomyces pombe Rho1p is essential, directly activates the (1,3)-ß-D-glucan synthase, and participates in regulation of cell wall growth and morphogenesis. Here we describe the characterization of the fission yeast Rho5p GTPase, highly homologous to Rho1p, sharing 86% identity and 95% similarity. Overexpression of the hyperactive allele rho5-G15V causes a morphological effect similar to that of rho1-G15V, but the penetrance is significantly lower, and overexpression of the dominant-negative allele rho5-T20N causes lysis like that of rho1-T20N. Importantly, overexpression of rho5⁺ but no other rho genes is able to rescue the lethality of rho1 cells. Shutoff experiments indicated that Rho5p can replace Rho1p, but it is not as effective in maintaining cell wall integrity or actin organization. rho5⁺ expression is hardly detected during log-phase growth but is induced under nutritional starvation conditions. rho5 cells are viable and do not display any defects during logarithmic growth. However, when rho1⁺ expression is repressed during stationary phase, rho5 cells display reduced viability. Ascospores lacking Rho5p are less resistant to heat or lytic enzymes than wild-type spores. Moreover, h⁹⁰ mutant strains carrying the hyperactive rho5-G15V or the dominant-negative rho5-T20N alleles display severe ascospore formation defects. These results suggest that Rho5p functions in a way similar to, but less efficient than, Rho1p, plays a nonessential role during stationary phase, and participates in the spore wall formation.

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