title: Proline biosynthesis is required for endoplasmic reticulum stress tolerance in Saccharomyces cerevisiae.
abstract: The amino acid proline is uniquely involved in cellular processes that underlie stress response in a variety of organisms. Proline is known to minimize protein aggregation, but a detailed study of how proline impacts cell survival during accumulation of misfolded proteins in the endoplasmic reticulum (ER) has not been performed. To address this we examined in Saccharomyces cerevisiae the effect of knocking out the PRO1, PRO2, and PRO3 genes responsible for proline biosynthesis. The null mutants pro1, pro2, and pro3 were shown to have increased sensitivity to ER stress relative to wild-type cells, which could be restored by proline or the corresponding genetic complementation. Of these mutants, pro3 was the most sensitive to tunicamycin and was rescued by anaerobic growth conditions or reduced thiol reagents. The pro3 mutant cells have higher intracellular reactive oxygen species, total glutathione, and a NADP(+)/NADPH ratio than wild-type cells under limiting proline conditions. Depletion of proline biosynthesis also inhibits the unfolded protein response (UPR) indicating proline protection involves the UPR. To more broadly test the role of proline in ER stress, increased proline biosynthesis was shown to partially rescue the ER stress sensitivity of a hog1 null mutant in which the high osmolality pathway is disrupted.
authors.affiliation: From the Department of Biochemistry and Redox Biology Center, University of Nebraska, Lincoln, Nebraska 68588 and.
authors.forename: Xinwen
authors.initials: X
authors.lastname: Liang
authors.affiliation: the Institute for Plant Genomics and Biotechnology, Department of Plant Pathology and Microbiology, Texas A&M University, College Station, Texas 77843.
authors.forename: Martin B
authors.initials: MB
authors.lastname: Dickman
authors.affiliation: From the Department of Biochemistry and Redox Biology Center, University of Nebraska, Lincoln, Nebraska 68588 and dbecker3@unl.edu.
authors.forename: Donald F
authors.initials: DF
authors.lastname: Becker
mesh_terms: D059865:Endoplasmic Reticulum Stress; D009249:NADP / Q000378:metabolism; D017851:Phosphotransferases (Carboxyl Group Acceptor) / Q000235:genetics / Q000378:metabolism; D011392:Proline / Q000096:biosynthesis*; D011762:Pyrroline Carboxylate Reductases / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D056811:Unfolded Protein Response; D000097502:delta-1-Pyrroline-5-Carboxylate Reductase
chemical_list: D009249:NADP; D017851:Phosphotransferases (Carboxyl Group Acceptor); D011392:Proline; D011762:Pyrroline Carboxylate Reductases; D000097502:delta-1-Pyrroline-5-Carboxylate Reductase; C020650:glutamate 5-kinase
keywords: Amino Acid; Biosynthesis; Endoplasmic Reticulum Stress (ER Stress); Glutathione; Metabolism; Proline; Redox
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