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On the illusion of auxotrophy: met15Δ yeast cells can grow on inorganic sulfur, thanks to the previously uncharacterized homocysteine synthase Yll058w.
Van Oss, S Branden; Parikh, Saurin Bipin; Castilho Coelho, Nelson; Wacholder, Aaron; Belashov, Ivan; Zdancewicz, Sara; Michaca, Manuel; Xu, Jiazhen; Kang, Yun Pyo; Ward, Nathan P; Yoon, Sang Jun; McCourt, Katherine M; McKee, Jake; Ideker, Trey; VanDemark, Andrew P; DeNicola, Gina M; Carvunis, Anne-Ruxandra.
Afiliación
  • Van Oss SB; Department of Computational and System Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA; Pittsburgh Center for Evolutionary Biology and Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Parikh SB; Department of Computational and System Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA; Pittsburgh Center for Evolutionary Biology and Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Castilho Coelho N; Department of Computational and System Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA; Pittsburgh Center for Evolutionary Biology and Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Wacholder A; Department of Computational and System Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA; Pittsburgh Center for Evolutionary Biology and Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Belashov I; Department of Biological Sciences, University of Pittsburgh, Dietrich School of Arts & Sciences, Pittsburgh, Pennsylvania, USA.
  • Zdancewicz S; Department of Biological Sciences, University of Pittsburgh, Dietrich School of Arts & Sciences, Pittsburgh, Pennsylvania, USA.
  • Michaca M; Department of Computational and System Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA; Pittsburgh Center for Evolutionary Biology and Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Xu J; Department of Computational and System Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Kang YP; Department of Cancer Physiology, H. Lee Moffitt Cancer Center, Tampa, Florida, USA.
  • Ward NP; Department of Cancer Physiology, H. Lee Moffitt Cancer Center, Tampa, Florida, USA.
  • Yoon SJ; Department of Cancer Physiology, H. Lee Moffitt Cancer Center, Tampa, Florida, USA.
  • McCourt KM; Department of Computational and System Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA; Pittsburgh Center for Evolutionary Biology and Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • McKee J; Department of Computational and System Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Ideker T; Departments of Medicine, Bioengineering, Computer Science and Engineering, Institute for Genomic Medicine, University of California San Diego, La Jolla, California, USA.
  • VanDemark AP; Department of Biological Sciences, University of Pittsburgh, Dietrich School of Arts & Sciences, Pittsburgh, Pennsylvania, USA.
  • DeNicola GM; Department of Cancer Physiology, H. Lee Moffitt Cancer Center, Tampa, Florida, USA.
  • Carvunis AR; Department of Computational and System Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA; Pittsburgh Center for Evolutionary Biology and Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA. Electronic address: anc201@pitt.edu.
J Biol Chem ; 298(12): 102697, 2022 12.
Article en En | MEDLINE | ID: mdl-36379252
Organisms must either synthesize or assimilate essential organic compounds to survive. The homocysteine synthase Met15 has been considered essential for inorganic sulfur assimilation in yeast since its discovery in the 1970s. As a result, MET15 has served as a genetic marker for hundreds of experiments that play a foundational role in eukaryote genetics and systems biology. Nevertheless, we demonstrate here through structural and evolutionary modeling, in vitro kinetic assays, and genetic complementation, that an alternative homocysteine synthase encoded by the previously uncharacterized gene YLL058W enables cells lacking Met15 to assimilate enough inorganic sulfur for survival and proliferation. These cells however fail to grow in patches or liquid cultures unless provided with exogenous methionine or other organosulfurs. We show that this growth failure, which has historically justified the status of MET15 as a classic auxotrophic marker, is largely explained by toxic accumulation of the gas hydrogen sulfide because of a metabolic bottleneck. When patched or cultured with a hydrogen sulfide chelator, and when propagated as colony grids, cells without Met15 assimilate inorganic sulfur and grow, and cells with Met15 achieve even higher yields. Thus, Met15 is not essential for inorganic sulfur assimilation in yeast. Instead, MET15 is the first example of a yeast gene whose loss conditionally prevents growth in a manner that depends on local gas exchange. Our results have broad implications for investigations of sulfur metabolism, including studies of stress response, methionine restriction, and aging. More generally, our findings illustrate how unappreciated experimental variables can obfuscate biological discovery.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Saccharomyces cerevisiae / Azufre / Proteínas de Saccharomyces cerevisiae Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: J Biol Chem Año: 2022 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Saccharomyces cerevisiae / Azufre / Proteínas de Saccharomyces cerevisiae Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: J Biol Chem Año: 2022 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos