Deciphering Microbial Metal Toxicity Responses using RB-TnSeq and Activity-Based Metabolomics
Abstract
To uncover metal toxicity targets and defense mechanisms of the facultative anaerobe Pantoea sp. strain MT58 (MT58), we used a multiomic strategy combining two global techniques, random bar code transposon site sequencing (RB-TnSeq) and activity-based metabolomics. MT58 is a metal-tolerant Oak Ridge Reservation (ORR) environmental isolate that was enriched in the presence of metals at concentrations measured in contaminated groundwater at an ORR nuclear waste site. The effects of three chemically different metals found at elevated concentrations in the ORR contaminated environment were investigated: the cation Al3+, the oxyanion CrO42-, and the oxycation UO22+. Both global techniques were applied using all three metals under both aerobic and anaerobic conditions to elucidate metal interactions mediated through the activity of metabolites and key genes/proteins. These revealed that Al3+ binds intracellular arginine, CrO42- enters the cell through sulfate transporters and oxidizes intracellular reduced thiols, and membrane-bound lipopolysaccharides protect the cell from UO22+ toxicity. In addition, the Tol outer membrane system contributed to the protection of cellular integrity from the toxic effects of all three metals. Likewise, we found evidence of regulation of lipid content in membranes under metal stress. Individually, RB-TnSeq and metabolomics are powerful tools to explore the impact various stresses havemore »
- Authors:
-
[3];
[1]
- Univ. of Georgia, Athens, GA (United States)
- Scripps Center for Mass Spectrometry and Metabolomics, San Diego, CA (United States); The Scripps Research Inst., La Jolla, CA (United States)
- Scripps Center for Mass Spectrometry and Metabolomics, San Diego, CA (United States); The Scripps Research Inst., La Jolla, CA (United States); Univ. of Wisconsin, Madison, WI (United States)
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1825256
- Grant/Contract Number:
- AC02-05CH11231
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Applied and Environmental Microbiology
- Additional Journal Information:
- Journal Volume: 87; Journal Issue: 21; Journal ID: ISSN 0099-2240
- Publisher:
- American Society for Microbiology
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Thorgersen, Michael P., Xue, Jingchuan, Majumder, Erica L.W., Trotter, Valentine V., Ge, Xiaoxuan, Poole, Farris L., Owens, Trenton K., Lui, Lauren M., Nielsen, Torben N., Arkin, Adam P., Deutschbauer, Adam M., Siuzdak, Gary, and Adams, Michael W. Deciphering Microbial Metal Toxicity Responses using RB-TnSeq and Activity-Based Metabolomics. United States: N. p., 2021.
Web. doi:10.1128/aem.01037-21.
Thorgersen, Michael P., Xue, Jingchuan, Majumder, Erica L.W., Trotter, Valentine V., Ge, Xiaoxuan, Poole, Farris L., Owens, Trenton K., Lui, Lauren M., Nielsen, Torben N., Arkin, Adam P., Deutschbauer, Adam M., Siuzdak, Gary, & Adams, Michael W. Deciphering Microbial Metal Toxicity Responses using RB-TnSeq and Activity-Based Metabolomics. United States. https://doi.org/10.1128/aem.01037-21
Thorgersen, Michael P., Xue, Jingchuan, Majumder, Erica L.W., Trotter, Valentine V., Ge, Xiaoxuan, Poole, Farris L., Owens, Trenton K., Lui, Lauren M., Nielsen, Torben N., Arkin, Adam P., Deutschbauer, Adam M., Siuzdak, Gary, and Adams, Michael W. Wed .
"Deciphering Microbial Metal Toxicity Responses using RB-TnSeq and Activity-Based Metabolomics". United States. https://doi.org/10.1128/aem.01037-21. https://www.osti.gov/servlets/purl/1825256.
@article{osti_1825256,
title = {Deciphering Microbial Metal Toxicity Responses using RB-TnSeq and Activity-Based Metabolomics},
author = {Thorgersen, Michael P. and Xue, Jingchuan and Majumder, Erica L.W. and Trotter, Valentine V. and Ge, Xiaoxuan and Poole, Farris L. and Owens, Trenton K. and Lui, Lauren M. and Nielsen, Torben N. and Arkin, Adam P. and Deutschbauer, Adam M. and Siuzdak, Gary and Adams, Michael W.},
abstractNote = {To uncover metal toxicity targets and defense mechanisms of the facultative anaerobe Pantoea sp. strain MT58 (MT58), we used a multiomic strategy combining two global techniques, random bar code transposon site sequencing (RB-TnSeq) and activity-based metabolomics. MT58 is a metal-tolerant Oak Ridge Reservation (ORR) environmental isolate that was enriched in the presence of metals at concentrations measured in contaminated groundwater at an ORR nuclear waste site. The effects of three chemically different metals found at elevated concentrations in the ORR contaminated environment were investigated: the cation Al3+, the oxyanion CrO42-, and the oxycation UO22+. Both global techniques were applied using all three metals under both aerobic and anaerobic conditions to elucidate metal interactions mediated through the activity of metabolites and key genes/proteins. These revealed that Al3+ binds intracellular arginine, CrO42- enters the cell through sulfate transporters and oxidizes intracellular reduced thiols, and membrane-bound lipopolysaccharides protect the cell from UO22+ toxicity. In addition, the Tol outer membrane system contributed to the protection of cellular integrity from the toxic effects of all three metals. Likewise, we found evidence of regulation of lipid content in membranes under metal stress. Individually, RB-TnSeq and metabolomics are powerful tools to explore the impact various stresses have on biological systems. In this work, we show that together they can be used synergistically to identify the molecular actors and mechanisms of these pertubations to an organism, furthering our understanding of how living systems interact with their environment.},
doi = {10.1128/aem.01037-21},
journal = {Applied and Environmental Microbiology},
number = 21,
volume = 87,
place = {United States},
year = {Wed Aug 25 00:00:00 EDT 2021},
month = {Wed Aug 25 00:00:00 EDT 2021}
}
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