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Large-scale investigation of oxygen response mutants in Saccharomyces cerevisiae.

Authors:
Samanfar B, Omidi K, Hooshyar M, Laliberte B, Alamgir M, Seal AJ, Ahmed-Muhsin E, Viteri DF, Said K, Chalabian F, Golshani A, Wainer G, Burnside D, Shostak K, Bugno M, Willmore WG, Smith ML, Golshani A
Affiliation:
Journal:
Molecular bioSystems

Abstract

A genome-wide screen of a yeast non-essential gene-deletion library was used to identify sick phenotypes due to oxygen deprivation. The screen provided a manageable list of 384 potentially novel as well as known oxygen responding (anoxia-survival) genes. The gene-deletion mutants were further assayed for sensitivity to ferrozine and cobalt to obtain a subset of 34 oxygen-responsive candidate genes including the known hypoxic gene activator, MGA2. With each mutant in this subset a plasmid based β-galactosidase assay was performed using the anoxic-inducible promoter from OLE1 gene, and 17 gene deletions were identified that inhibit induction under anaerobic conditions. Genetic interaction analysis for one of these mutants, the RNase-encoding POP2 gene, revealed synthetic sick interactions with a number of genes involved in oxygen sensing and response. Knockdown experiments for CNOT8, human homolog of POP2, reduced cell survival under low oxygen condition suggesting a similar function in human cells.

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