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High-throughput interaction screens illuminate the role of c-di-AMP in cyanobacterial nighttime survival.
Rubin, Benjamin E; Huynh, TuAnh Ngoc; Welkie, David G; Diamond, Spencer; Simkovsky, Ryan; Pierce, Emily C; Taton, Arnaud; Lowe, Laura C; Lee, Jenny J; Rifkin, Scott A; Woodward, Joshua J; Golden, Susan S.
Afiliación
  • Rubin BE; Division of Biological Sciences, University of California San Diego, La Jolla, CA, United States of America.
  • Huynh TN; Department of Microbiology, University of Washington, Seattle, WA, United States of America.
  • Welkie DG; Center for Circadian Biology, University of California San Diego, La Jolla, CA, United States of America.
  • Diamond S; Division of Biological Sciences, University of California San Diego, La Jolla, CA, United States of America.
  • Simkovsky R; Center for Circadian Biology, University of California San Diego, La Jolla, CA, United States of America.
  • Pierce EC; Division of Biological Sciences, University of California San Diego, La Jolla, CA, United States of America.
  • Taton A; Division of Biological Sciences, University of California San Diego, La Jolla, CA, United States of America.
  • Lowe LC; Center for Circadian Biology, University of California San Diego, La Jolla, CA, United States of America.
  • Lee JJ; Division of Biological Sciences, University of California San Diego, La Jolla, CA, United States of America.
  • Rifkin SA; Division of Biological Sciences, University of California San Diego, La Jolla, CA, United States of America.
  • Woodward JJ; Department of Microbiology, University of Washington, Seattle, WA, United States of America.
  • Golden SS; Division of Biological Sciences, University of California San Diego, La Jolla, CA, United States of America.
PLoS Genet ; 14(4): e1007301, 2018 04.
Article en En | MEDLINE | ID: mdl-29608558
The broadly conserved signaling nucleotide cyclic di-adenosine monophosphate (c-di-AMP) is essential for viability in most bacteria where it has been studied. However, characterization of the cellular functions and metabolism of c-di-AMP has largely been confined to the class Bacilli, limiting our functional understanding of the molecule among diverse phyla. We identified the cyclase responsible for c-di-AMP synthesis and characterized the molecule's role in survival of darkness in the model photosynthetic cyanobacterium Synechococcus elongatus PCC 7942. In addition to the use of traditional genetic, biochemical, and proteomic approaches, we developed a high-throughput genetic interaction screen (IRB-Seq) to determine pathways where the signaling nucleotide is active. We found that in S. elongatus c-di-AMP is produced by an enzyme of the diadenylate cyclase family, CdaA, which was previously unexplored experimentally. A cdaA-null mutant experiences increased oxidative stress and death during the nighttime portion of day-night cycles, in which potassium transport is implicated. These findings suggest that c-di-AMP is biologically active in cyanobacteria and has non-canonical roles in the phylum including oxidative stress management and day-night survival. The pipeline and analysis tools for IRB-Seq developed for this study constitute a quantitative high-throughput approach for studying genetic interactions.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: AMP Cíclico / Synechococcus / Ensayos Analíticos de Alto Rendimiento Tipo de estudio: Prognostic_studies Idioma: En Revista: PLoS Genet Asunto de la revista: GENETICA Año: 2018 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: AMP Cíclico / Synechococcus / Ensayos Analíticos de Alto Rendimiento Tipo de estudio: Prognostic_studies Idioma: En Revista: PLoS Genet Asunto de la revista: GENETICA Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos