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Title: Tn-seq of Caulobacter crescentus under uranium stress reveals genes essential for detoxification and stress tolerance

Abstract

Ubiquitous aquatic bacterium Caulobacter crescentus is highly resistant to uranium (U) and facilitates U biomineralization and thus holds promise as an agent of U bioremediation. In order to gain an understanding of how C. crescentus tolerates U, we employed transposon (Tn) mutagenesis paired with deep sequencing (Tn-seq) in a global screen for genomic elements required for U resistance. Of the 3,879 annotated genes in the C. crescentus genome, 37 were found to be specifically associated with fitness under U stress, 15 of which were subsequently tested through mutational analysis. Systematic deletion analysis revealed that mutants lacking outer membrane transporters (rsaFa and rsaFb), a stress-responsive transcription factor (cztR), or a ppGpp synthetase/hydrolase (spoT) exhibited a significantly lower survival rate under U stress. RsaFa and RsaFb, which are homologues of TolC in Escherichia coli, have previously been shown to mediate S-layer export. Transcriptional analysis revealed upregulation of rsaFa and rsaFb by 4- and 10-fold, respectively, in the presence of U. We additionally show that rsaFa mutants accumulated higher levels of U than the wild type, with no significant increase in oxidative stress levels. These results suggest a function for RsaFa and RsaFb in U efflux and/or maintenance of membrane integrity during Umore » stress. In addition, we present data implicating CztR and SpoT in resistance to U stress. Together, our findings reveal novel gene targets that are key to understanding the molecular mechanisms of U resistance in C. crescentus.« less

Authors:
 [1];  [1];  [1];  [2];  [2];  [3];  [4];  [5];  [6];  [7];  [1]
  1. Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
  2. DOE Joint Genome Inst., Walnut Creek, CA (United States)
  3. Univ. of British Columbia, Vancouver, BC (Canada)
  4. California State Univ., Northridge, CA (United States)
  5. Stanford Univ. School of Medicine, Stanford, CA (United States)
  6. ETH Zurich (Switzerland)
  7. Univ. of Wyoming, Laramie, WY (United States)
Publication Date:
Research Org.:
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1234593
Report Number(s):
LLNL-JRNL-670205
Journal ID: ISSN 0021-9193; TRN: US1600611
Grant/Contract Number:  
AC52-07NA27344
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Bacteriology
Additional Journal Information:
Journal Volume: 197; Journal Issue: 19; Journal ID: ISSN 0021-9193
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
63 RADIATION, THERMAL, AND OTHER ENVIRONMENTAL POLLUTANT EFFECTS ON LIVING ORGANISMS AND BIOLOGICAL MATERIALS; 59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Yung, Mimi C., Park, Dan M., Overton, K. Wesley, Blow, Matthew J., Hoover, Cindi A., Smit, John R., Murray, Sean R., Ricci, Dante P., Christen, Beat, Bowman, Grant R., and Jiao, Yongqin. Tn-seq of Caulobacter crescentus under uranium stress reveals genes essential for detoxification and stress tolerance. United States: N. p., 2015. Web. doi:10.1128/JB.00382-15.
Yung, Mimi C., Park, Dan M., Overton, K. Wesley, Blow, Matthew J., Hoover, Cindi A., Smit, John R., Murray, Sean R., Ricci, Dante P., Christen, Beat, Bowman, Grant R., & Jiao, Yongqin. Tn-seq of Caulobacter crescentus under uranium stress reveals genes essential for detoxification and stress tolerance. United States. https://doi.org/10.1128/JB.00382-15
Yung, Mimi C., Park, Dan M., Overton, K. Wesley, Blow, Matthew J., Hoover, Cindi A., Smit, John R., Murray, Sean R., Ricci, Dante P., Christen, Beat, Bowman, Grant R., and Jiao, Yongqin. Mon . "Tn-seq of Caulobacter crescentus under uranium stress reveals genes essential for detoxification and stress tolerance". United States. https://doi.org/10.1128/JB.00382-15. https://www.osti.gov/servlets/purl/1234593.
@article{osti_1234593,
title = {Tn-seq of Caulobacter crescentus under uranium stress reveals genes essential for detoxification and stress tolerance},
author = {Yung, Mimi C. and Park, Dan M. and Overton, K. Wesley and Blow, Matthew J. and Hoover, Cindi A. and Smit, John R. and Murray, Sean R. and Ricci, Dante P. and Christen, Beat and Bowman, Grant R. and Jiao, Yongqin},
abstractNote = {Ubiquitous aquatic bacterium Caulobacter crescentus is highly resistant to uranium (U) and facilitates U biomineralization and thus holds promise as an agent of U bioremediation. In order to gain an understanding of how C. crescentus tolerates U, we employed transposon (Tn) mutagenesis paired with deep sequencing (Tn-seq) in a global screen for genomic elements required for U resistance. Of the 3,879 annotated genes in the C. crescentus genome, 37 were found to be specifically associated with fitness under U stress, 15 of which were subsequently tested through mutational analysis. Systematic deletion analysis revealed that mutants lacking outer membrane transporters (rsaFa and rsaFb), a stress-responsive transcription factor (cztR), or a ppGpp synthetase/hydrolase (spoT) exhibited a significantly lower survival rate under U stress. RsaFa and RsaFb, which are homologues of TolC in Escherichia coli, have previously been shown to mediate S-layer export. Transcriptional analysis revealed upregulation of rsaFa and rsaFb by 4- and 10-fold, respectively, in the presence of U. We additionally show that rsaFa mutants accumulated higher levels of U than the wild type, with no significant increase in oxidative stress levels. These results suggest a function for RsaFa and RsaFb in U efflux and/or maintenance of membrane integrity during U stress. In addition, we present data implicating CztR and SpoT in resistance to U stress. Together, our findings reveal novel gene targets that are key to understanding the molecular mechanisms of U resistance in C. crescentus.},
doi = {10.1128/JB.00382-15},
journal = {Journal of Bacteriology},
number = 19,
volume = 197,
place = {United States},
year = {Mon Jul 20 00:00:00 EDT 2015},
month = {Mon Jul 20 00:00:00 EDT 2015}
}

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