title: Metabolic Regulation as a Consequence of Anaerobic 5-Methylthioadenosine Recycling in Rhodospirillum rubrum.
abstract: Rhodospirillum rubrum possesses a novel oxygen-independent, aerobic methionine salvage pathway (MSP) for recycling methionine from 5-methylthioadenosine (MTA), the MTA-isoprenoid shunt. This organism can also metabolize MTA as a sulfur source under anaerobic conditions, suggesting that the MTA-isoprenoid shunt may also function anaerobically as well. In this study, deep proteomics profiling, directed metabolite analysis, and reverse transcriptase quantitative PCR (RT-qPCR) revealed metabolic changes in response to anaerobic growth on MTA versus sulfate as sole sulfur source. The abundance of protein levels associated with methionine transport, cell motility, and chemotaxis increased in the presence of MTA over that in the presence of sulfate. Purine salvage from MTA resulted primarily in hypoxanthine accumulation and a decrease in protein levels involved in GMP-to-AMP conversion to balance purine pools. Acyl coenzyme A (acyl-CoA) metabolic protein levels for lipid metabolism were lower in abundance, whereas poly-β-hydroxybutyrate synthesis and storage were increased nearly 10-fold. The known R. rubrum aerobic MSP was also shown to be upregulated, to function anaerobically, and to recycle MTA. This suggested that other organisms with gene homologues for the MTA-isoprenoid shunt may also possess a functioning anaerobic MSP. In support of our previous findings that ribulose-1,5-carboxylase/oxygenase (RubisCO) is required for an apparently purely anaerobic MSP, RubisCO transcript and protein levels both increased in abundance by over 10-fold in cells grown anaerobically on MTA over those in cells grown on sulfate, resulting in increased intracellular RubisCO activity. These results reveal for the first time global metabolic responses as a consequence of anaerobic MTA metabolism compared to using sulfate as the sulfur source.
  
  
  
  In nearly all organisms, sulfur-containing byproducts result from many metabolic reactions. Unless these compounds are further metabolized, valuable organic sulfur is lost and can become limiting. To regenerate the sulfur-containing amino acid methionine, organisms typically employ one of several variations of a "universal" methionine salvage pathway (MSP). A common aspect of the universal MSP is a final oxygenation step. This work establishes that the metabolically versatile bacterium Rhodospirillum rubrum employs a novel MSP that does not require oxygen under either aerobic or anaerobic conditions. There is also a separate, dedicated anaerobic MTA metabolic route in R. rubrum This work reveals global changes in cellular metabolism in response to anaerobic MTA metabolism compared to using sulfate as a sulfur source. We found that cell mobility and transport were enhanced, along with lipid, nucleotide, and carbohydrate metabolism, when cells were grown in the presence of MTA.
authors.affiliation: Department of Microbiology, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Justin A
authors.initials: JA
authors.lastname: North
authors.affiliation: Department of Microbiology, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Jaya
authors.initials: J
authors.lastname: Sriram
authors.affiliation: Chemical Sciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA.
authors.forename: Karuna
authors.initials: K
authors.lastname: Chourey
authors.affiliation: Department of Microbiology, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Ecker
authors.affiliation: University of Tennessee-ORNL Graduate School of Genome Science and Technology, Knoxville, Tennessee, USA.
authors.forename: Ritin
authors.initials: R
authors.lastname: Sharma
authors.affiliation: Department of Microbiology, The Ohio State University, Columbus, Ohio, USA.
authors.forename: John A
authors.initials: JA
authors.lastname: Wildenthal
authors.affiliation: Chemical Sciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Hettich
authors.affiliation: Department of Microbiology, The Ohio State University, Columbus, Ohio, USA tabita.1@osu.edu.
authors.forename: F Robert
authors.initials: FR
authors.lastname: Tabita
mesh_terms: D000332:Aerobiosis; D000693:Anaerobiosis; D003839:Deoxyadenosines / Q000378:metabolism*; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects*; D053858:Metabolic Networks and Pathways; D055442:Metabolome; D020543:Proteome / Q000032:analysis; D012247:Rhodospirillum rubrum / Q000254:growth & development / Q000378:metabolism*; D013872:Thionucleosides / Q000378:metabolism*
chemical_list: D003839:Deoxyadenosines; D020543:Proteome; D013872:Thionucleosides; C008500:5'-methylthioadenosine