Deciphering Microbial Metal Toxicity Responses using RB-TnSeq and Activity-Based Metabolomics

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.

doi:10.1128/aem.01037-21

Title: Deciphering Microbial Metal Toxicity Responses using RB-TnSeq and Activity-Based Metabolomics

Journal Article · 24 August 2021 · Applied and Environmental Microbiology

DOI: https://doi.org/10.1128/aem.01037-21 · OSTI ID:1825256

Thorgersen, Michael P. ^[1]; Xue, Jingchuan ^[2];

^[3]; Trotter, Valentine V. ^[4]; Ge, Xiaoxuan ^[1]; Poole, Farris L. ^[1]; Owens, Trenton K. ^[4]; Lui, Lauren M. ^[4]; Nielsen, Torben N. ^[4]; Arkin, Adam P. ^[4]; Deutschbauer, Adam M. ^[4]; Siuzdak, Gary ^[2];

^[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)

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 CrO₄^2-, 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, CrO₄^2- 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.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1825256

Journal Information:: Applied and Environmental Microbiology, Journal Name: Applied and Environmental Microbiology Journal Issue: 21 Vol. 87; ISSN 0099-2240

Publisher:: American Society for MicrobiologyCopyright Statement

Country of Publication:: United States

Language:: English

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