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Applied Microbiology and Biotechnology

, Volume 103, Issue 6, pp 2701–2713 | Cite as

Gene function and expression regulation of RuvRCAB in bacterial Cr(VI), As(III), Sb(III), and Cd(II) resistance

  • Shijuan Wu
  • Xian Xia
  • Dan Wang
  • Zijie Zhou
  • Gejiao WangEmail author
Applied genetics and molecular biotechnology
  • 236 Downloads

Abstract

Alishewanella sp. WH16-1 is a heavy metal-resistant bacterium. Previously, a putative YebC family regulator gene, designated ruvR, was associated with Cr(VI) resistance. In this study, comprehensive analyses were performed to study the role of ruvR and its adjunct putative DNA repairing genes, ruvCAB, in the heavy metal resistance of Alishewanella sp. WH16-1. RT-PCR analysis showed that ruvR is cotranscribed with ruvCAB. Gene mutation and complementation experiments indicated that ruvRCAB contributes to Cr(VI), As(III), Sb(III), and Cd(II) resistance in vivo. Random amplification of polymorphic DNA-PCR revealed that ruvCAB is associated with DNA repair mediated by these metals, and the presence of the metals in the cells was confirmed by elemental mapping and energy-dispersive X-ray spectrograph analysis. In addition, qRT-PCR, reporter gene assay, and in vivo and in vitro protein-DNA interaction experiments indicated that RuvR positively regulates the transcription of ruvCAB and is induced by Cr(VI). Finally, site-directed mutagenesis demonstrated that Asp103 is essential for the DNA binding ability of RuvR. The above results suggest that RuvR is involved in Cr(VI) resistance and resistance to other metals and that RuvR positively regulates the expression of ruvCAB. Based on our study and literatures, a model of RuvRCAB detailing bacterial heavy metal resistance is proposed. The RuvRCAB system plays an important role in the ability of Alishewanella sp. WH16-1 to survive in environments with heavy metals.

Keywords

Alishewanella Chromate DNA repair proteins RuvCAB Metal(loid) resistance RuvR (YebC family protein) 

Notes

Funding

This study was funded by the National Key Research and Development Program of China (grant number 2016YFD0800702) and National Natural Science Foundation of China (grant number 31870086).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2019_9666_MOESM1_ESM.pdf (1.5 mb)
ESM 1 (PDF 1530 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Shijuan Wu
    • 1
  • Xian Xia
    • 1
  • Dan Wang
    • 1
  • Zijie Zhou
    • 1
  • Gejiao Wang
    • 1
    Email author
  1. 1.State Key Laboratory of Agricultural Microbiology, College of Life Science and TechnologyHuazhong Agricultural UniversityWuhanPeople’s Republic of China

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