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(The FASEB Journal. 2006;20:A1348)
© 2006 FASEB
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867.19

Functional Screening of the Saccharomyces cerevisiae Deletion Collection for Ino– and Opi– Phenotypes

Flora Irma Hinz, Myung Sun Choi, Manuel J. Villa-García and Susan A. Henry

Department of Molecular Biology and Genetics, Cornell University, 245 Biotechnology Building, Ithaca, NY, 14853

ABSTRACT

In Saccharomyces cerevisiae, co-regulated genes involved in phospholipid biosynthesis which contain the repeated cis acting element, UASINO, in their promoters, are coordinately repressed when cells are grown in media containing the phospholipid precursors, inositol and choline. The most highly regulated of the UASINO containing genes is INO1, the structural gene for myo-inositol-3-phosphate synthase. Constitutive expression of INO1 results in overproduction of inositol (Opi– phenotype), while failure to express INO1 leads to inositol auxotrophy (Ino– phenotype). We report the screening at two temperatures of an entire collection of homozygous diploid yeast deletion strains (approx. 4600 strains) following growth on media containing or lacking inositol or choline. Validating this approach, previously characterized mutants presenting Ino– or Opi– phenotypes were readily detected using our screening method. Numerous previously uncharacterized Ino– mutants, and several new Opi– mutants, were identified. These new categories of mutants are providing new avenues for future research as well as novel insights into regulatory networks connecting phospholipid metabolism to diverse aspects of cellular signalling and metabolism.

Supported by a grant of the National Institutes of Health to S.A.H. (GM019629-31).





This Article
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Right arrow Articles by Hinz, F. I.
Right arrow Articles by Henry, S. A.