Initial molecular characterization of a novel Rho-type GTPase RhoH in the filamentous ascomycete Ashbya gossypii.

The organization of the actin cytoskeleton is of central importance in determining cell shape and morphogenesis. Rho-type GTP-binding proteins are known for their role to activate downstream effector proteins that act as scaffolds and direct the assembly of actin filaments. We have identified a novel Rho-type GTPase, RHOH, in the filamentous ascomycete Ashbya gossypii and also in one of its closest relatives, Holleya sinecauda both species belonging to the genus Eremothecium. RHOH is a paralog of AgRHO1 and was generated by tandem duplication in an evolutionary ancestor but is not found in Saccharomyces cerevisae. Deletion of RHOH was carried out in both A. gossypii and H. sinecauda and showed that this gene is non-essential. However, a cell wall defect was observed that led to increased cell lysis at the hyphal tip and an increased sensitivity against the cell wall agents calcofluor and SDS in the AgrhoH strain. These results suggested at least partially overlapping functions between RhoH and Rho1 without excluding any spilt functions of these proteins. Deletion of both the RHOH and RHO1 genes in A. gossypii exacerbated the mutant rho1 phenotype and resulted in germination deficient spores. This suggests that Rho1/RhoH -in contrast to Cdc42- are required at the early steps of spore germination.