Synthetic genetic and phenotypic analysis of gene modules involved in chitin synthesis.
Guillaume Lesage (1), Jesse Shapiro (1), Charles Specht (2), Arnaud Firon (1), Anne-Marie Sdicu (1), Patrice Menard (1), Shamiza Hussein (1), Amy H.Y. Tong (3), Charles Boone (3), Howard Bussey (1)
(1) Department of Biology, McGill University, 1205 Dr. Penfield, Montreal, QC, H3A 1B1, Canada;
(2) Department of Medicine, Boston University, Boston (MA);
(3) Banting and Best Department of Medical Research, University of Toronto, Toronto (ON)
In S. cerevisiae, chitin, a linear polymer of beta-1, 4-linked N-acetylglucosamine, is synthesized by three chitin synthases (Chs1p, Chs2p and Chs3p). To find genes buffering against mutations leading to chitin defects, we performed screens for synthetic lethal/sick interactions with genes encoding chitin synthases I and III (CHS1 and CHS3) and for genes required for the optimal activity or localization of Chs3p (CHS4, CHS5, CHS6, CHS7 and BNI4). A genetic network was found which, in addition to reconstructing much of what is known of chitin biology, implicates many new genes in this process. To extend the analysis, we quantified total chitin levels and assayed the calcofluor white sensitivity of mutants in the chitin network. By grouping interacting genes according to their mutant phenotypes we could (i) make some specific functional predictions and (ii) place chitin synthesis in a global cellular context.
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