title: Label-free and redox proteomic analyses of the triacylglycerol-accumulating Rhodococcus jostii RHA1.
abstract: The bacterium Rhodococcus jostii RHA1 synthesizes large amounts of triacylglycerols (TAGs) under conditions of nitrogen starvation. To better understand the molecular mechanisms behind this process, we performed proteomic studies in this oleaginous bacterium. Upon nitrogen starvation, we observed a re-routing of the carbon flux towards the formation of TAGs. Under these conditions, the cellular lipid content made up more than half of the cell's dry weight. On the proteome level, this coincided with a shift towards non-glycolytic carbohydrate-metabolizing pathways. These pathways (Entner-Doudoroff and pentose-phosphate shunt) contribute NADPH and precursors of glycerol 3-phosphate and acetyl-CoA to lipogenesis. The expression of proteins involved in the degradation of branched-chain amino acids and the methylmalonyl-CoA pathway probably provided propionyl-CoA for the biosynthesis of odd-numbered fatty acids, which make up almost 30 % of RHA1 fatty acid composition. Additionally, lipolytic and glycerol-degrading enzymes increased in abundance, suggesting a dynamic cycling of cellular lipids. Conversely, abundance of proteins involved in consuming intermediates of lipogenesis decreased. Furthermore, we identified another level of lipogenesis regulation through redox-mediated thiol modification in R. jostii. Enzymes affected included acetyl-CoA carboxylase and a β-ketoacyl-[acyl-carrier protein] synthase II (FabF). An integrative metabolic model for the oleaginous RHA1 strain is proposed on the basis of our results.
authors.affiliation: Centro Regional de Investigación y Desarrollo Científico Tecnológico, Facultad de Ciencias Naturales, Universidad Nacional de la Patagonia, San Juan Bosco Km 4-Ciudad Universitaria, 9000 Comodoro Rivadavia (Chubut), Argentina.
authors.forename: José Sebastián
authors.initials: JS
authors.lastname: Dávila Costa
authors.affiliation: Centro Regional de Investigación y Desarrollo Científico Tecnológico, Facultad de Ciencias Naturales, Universidad Nacional de la Patagonia, San Juan Bosco Km 4-Ciudad Universitaria, 9000 Comodoro Rivadavia (Chubut), Argentina Oil m&s, Av. Hipólito Yrigoyen 4250, 9000 Comodoro Rivadavia (Chubut), Argentina.
authors.forename: O Marisa
authors.initials: OM
authors.lastname: Herrero
authors.affiliation: Centro Regional de Investigación y Desarrollo Científico Tecnológico, Facultad de Ciencias Naturales, Universidad Nacional de la Patagonia, San Juan Bosco Km 4-Ciudad Universitaria, 9000 Comodoro Rivadavia (Chubut), Argentina halvarez@unpata.edu.ar lars.leichert@ruhr-uni-bochum.de.
authors.forename: Héctor M
authors.initials: HM
authors.lastname: Alvarez
authors.affiliation: Ruhr-Universitat Bochum, Medizinisches Proteom-Center, Redox-Proteomics Group, Bochum, Germany halvarez@unpata.edu.ar lars.leichert@ruhr-uni-bochum.de.
authors.forename: Lars
authors.initials: L
authors.lastname: Leichert
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism; D010084:Oxidation-Reduction; D040901:Proteomics; D012240:Rhodococcus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014280:Triglycerides / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D005227:Fatty Acids; D014280:Triglycerides