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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 purelymore » 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. Lastly, 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.« less

Authors:
 [1];  [1];  [2];  [1];  [3];  [1];  [2];  [1]
  1. The Ohio State Univ., Columbus, OH (United States)
  2. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  3. Univ. of Tennessee, Knoxville, TN (United States). ORNL Graduate School of Genome Science and Tech.
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1326542
Grant/Contract Number:  
AC05-00OR22725
Resource Type:
Accepted Manuscript
Journal Name:
mBio (Online)
Additional Journal Information:
Journal Name: mBio (Online); Journal Volume: 7; Journal Issue: 4; Journal ID: ISSN 2150-7511
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

North, Justin A., Sriram, Jaya, Chourey, Karuna, Ecker, Christopher D., Sharma, Ritin, Wildenthal, John A., Hettich, Robert L., and Tabita, F. Robert. Metabolic regulation as a consequence of anaerobic 5-methylthioadenosine recycling in Rhodospirillum rubrum. United States: N. p., 2016. Web. doi:10.1128/mBio.00855-16.
North, Justin A., Sriram, Jaya, Chourey, Karuna, Ecker, Christopher D., Sharma, Ritin, Wildenthal, John A., Hettich, Robert L., & Tabita, F. Robert. Metabolic regulation as a consequence of anaerobic 5-methylthioadenosine recycling in Rhodospirillum rubrum. United States. doi:10.1128/mBio.00855-16.
North, Justin A., Sriram, Jaya, Chourey, Karuna, Ecker, Christopher D., Sharma, Ritin, Wildenthal, John A., Hettich, Robert L., and Tabita, F. Robert. Tue . "Metabolic regulation as a consequence of anaerobic 5-methylthioadenosine recycling in Rhodospirillum rubrum". United States. doi:10.1128/mBio.00855-16. https://www.osti.gov/servlets/purl/1326542.
@article{osti_1326542,
title = {Metabolic regulation as a consequence of anaerobic 5-methylthioadenosine recycling in Rhodospirillum rubrum},
author = {North, Justin A. and Sriram, Jaya and Chourey, Karuna and Ecker, Christopher D. and Sharma, Ritin and Wildenthal, John A. and Hettich, Robert L. and Tabita, F. Robert},
abstractNote = {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. Lastly, 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.},
doi = {10.1128/mBio.00855-16},
journal = {mBio (Online)},
number = 4,
volume = 7,
place = {United States},
year = {2016},
month = {7}
}

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