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Genetics, Vol. 162, 663-676, October 2002, Copyright © 2002

Dissection of Upstream Regulatory Components of the Rho1p Effector, 1,3-ß-Glucan Synthase, in Saccharomyces cerevisiae

Mariko Sekiya-Kawasakia, Mitsuhiro Abea, Ayaka Sakaa, Daisuke Watanabea, Keiko Konoa, Masayo Minemura-Asakawaa, Satoru Ishiharaa, Takahide Watanabeb, and Yoshikazu Ohyaa,c
a Department of Integrated Biosciences, Graduate School of Frontier Science, University of Tokyo, Kashiwanoha, Kashiwa, Chiba 277-8562, Japan,
b Department of Mycology, Nippon Roche Research Center, Kajiwara, Kamakura, Kanagawa 247-8530, Japan
c Unit Process and Combined Circuit, PRESTO, Japan Science and Technology Corporation, Shinsenri-Higashimachi, Toyonaka, Osaka 565-0082, Japan

Corresponding author: Yoshikazu Ohya, Graduate School of Frontier Science, University of Tokyo, Bldg. FSB-101, 5-1-5 Kashiwanoha, Kashiwa, Chiba 277-8562, Japan., ohya{at}k.u-tokyo.ac.jp (E-mail)

Communicating editor: M. D. ROSE

In the budding yeast Saccharomyces cerevisiae, one of the main structural components of the cell wall is 1,3-ß-glucan produced by 1,3-ß-glucan synthase (GS). Yeast GS is composed of a putative catalytic subunit encoded by FKS1 and FKS2 and a regulatory subunit encoded by RHO1. A combination of amino acid alterations in the putative catalytic domain of Fks1p was found to result in a loss of the catalytic activity. To identify upstream regulators of 1,3-ß-glucan synthesis, we isolated multicopy suppressors of the GS mutation. We demonstrate that all of the multicopy suppressors obtained (WSC1, WSC3, MTL1, ROM2, LRE1, ZDS1, and MSB1) and the constitutively active RHO1 mutations tested restore 1,3-ß-glucan synthesis in the GS mutant. A deletion of either ROM2 or WSC1 leads to a significant defect of 1,3-ß-glucan synthesis. Analyses of the degree of Mpk1p phosphorylation revealed that among the multicopy suppressors, WSC1, ROM2, LRE1, MSB1, and MTL1 act positively on the Pkc1p-MAPK pathway, another signaling pathway regulated by Rho1p, while WSC3 and ZDS1 do not. We have also found that MID2 acts positively on Pkc1p without affecting 1,3-ß-glucan synthesis. These results suggest that distinct networks regulate the two effector proteins of Rho1p, Fks1p and Pkc1p.




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