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Mutations that are synthetically lethal with a gas1Δ allele cause defects in the cell wall of Saccharomyces cerevisiae

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Abstract

The GAS1 -related genes of fungi encode GPI-anchored proteins with β-1,3-glucanosyltransferase activity. Loss of this activity results in defects in the assembly of the cell wall. We isolated mutants that show a synthetic defect when combined with a gas1Δ allele in Saccharomyces cerevisiae, and identified nine wild-type genes that rescue this defect. The indispensability of BIG1 and KRE6 for the viability of gas1Δ cells confirmed the important role of β-1,6-glucan in cells that are defective in the processing of β-1,3-glucan. The identification of the Wsc1p hypo-osmotic stress sensor and components of the PKC signal transduction pathway in our screen also confirmed that the cell wall integrity response attenuates the otherwise lethal gas1Δ defect. Unexpectedly, we found that the KEX2 gene is also required for the viability of the gas1Δ mutant. Kex2p is a Golgi/endosome-membrane-anchored protease that processes secretory preproteins. A cell wall defect was also found in the kex2Δ mutant, which was suppressible by multiple copies of the MKC7 or YAP3 gene, both of which encode other GPI-anchored proteases. Therefore, normal cell wall assembly requires proteolytic processing of secretory preproteins. Furthermore, the genes CSG2 and IPT1 were found to be required for normal growth of gas1Δ cells in the presence of 1 M sorbitol. This finding suggests that complex sphingolipids play a role in the hyper-osmotic response.

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Acknowledgements

We would like to thank Akio Toh-e (University of Tokyo) for yeast strains and plasmids, Yves Bourbonnais (Université Laval) for antibody against Yap3p, Hiroshi Kitagaki (the National Research Institute of Brewing) for his valuable advice in determination of the Man/Glc ratio, and Ikuko Sato (Fujisawa Pharmaceutical Co.) for her contribution of K1 killer toxin and valuable advice. This work was supported by a grant-in-aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan, grants for Bioarchitecture Research from the Institute of Physical and Chemical Research (RIKEN), a grant from the Kato Memorial Bioscience Foundation (to Y. N.), and a grant from the Nagase Science and Technology Foundation (to K. Y.)

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Authors and Affiliations

  1. Department of Biotechnology, University of Tokyo, Bunkyo-ku, 113-8657, Tokyo, Japan

    N. Tomishige, Y. Noda, H. Adachi & K. Yoda

  2. National Research Institute of Brewing, 739-0046, Higashi-Hiroshima, Hiroshima, Japan

    H. Shimoi

  3. Institute of Physical and Chemical Research (RIKEN), 351-0198, Wako, Saitama, Japan

    A. Takatsuki

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  1. N. Tomishige
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  2. Y. Noda
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  3. H. Adachi
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  4. H. Shimoi
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  6. K. Yoda
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Correspondence to K. Yoda.

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Communicated by C. P. Hollenberg

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Tomishige, N., Noda, Y., Adachi, H. et al. Mutations that are synthetically lethal with a gas1Δ allele cause defects in the cell wall of Saccharomyces cerevisiae . Mol Gen Genomics 269, 562–573 (2003). https://doi.org/10.1007/s00438-003-0864-9

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