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Originally published as Genetics Published Articles Ahead of Print on April 2, 2006.

Genetics, Vol. 173, 621-634, June 2006, Copyright © 2006
doi:10.1534/genetics.106.057489

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Genomic Analysis of the Opi Phenotype

Leandria C. Hancock, Ryan P. Behta and John M. Lopes1

Department of Biological Sciences, Wayne State University, Detroit, Michigan 48202

1 Corresponding author: Department of Biological Sciences, Wayne State University, 5047 Gullen Mall, Detroit, MI 48202.
E-mail: jlopes{at}sun.science.wayne.edu

Most of the phospholipid biosynthetic genes of Saccharomyces cerevisiae are coordinately regulated in response to inositol and choline. Inositol affects the intracellular levels of phosphatidic acid (PA). Opi1p is a repressor of the phospholipid biosynthetic genes and specifically binds PA in the endoplasmic reticulum. In the presence of inositol, PA levels decrease, releasing Opi1p into the nucleus where it represses transcription. The opi1 mutant overproduces and excretes inositol into the growth medium in the absence of inositol and choline (Opi phenotype). To better understand the mechanism of Opi1p repression, the viable yeast deletion set was screened to identify Opi mutants. In total, 89 Opi mutants were identified, of which 7 were previously known to have the Opi phenotype. The Opi mutant collection included genes with roles in phospholipid biosynthesis, transcription, protein processing/synthesis, and protein trafficking. Included in this set were all nonessential components of the NuA4 HAT complex and six proteins in the Rpd3p–Sin3p HDAC complex. It has previously been shown that defects in phosphatidylcholine synthesis (cho2 and opi3) yield the Opi phenotype because of a buildup of PA. However, in this case the Opi phenotype is conditional because PA can be shuttled through a salvage pathway (Kennedy pathway) by adding choline to the growth medium. Seven new mutants present in the Opi collection (fun26, kex1, nup84, tps1, mrpl38, mrpl49, and opi10/yol032w) were also suppressed by choline, suggesting that these affect PC synthesis. Regulation in response to inositol is also coordinated with the unfolded protein response (UPR). Consistent with this, several Opi mutants were found to affect the UPR (yhi9, ede1, and vps74).







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Copyright © 2006 by the Genetics Society of America.