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Characterization of pbt genes conferring increased Pb(2+) and Cd(2+) tolerance upon Achromobacter xylosoxidans A8
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SYSNO ASEP 0422943 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Characterization of pbt genes conferring increased Pb(2+) and Cd(2+) tolerance upon Achromobacter xylosoxidans A8 Author(s) Hložková, K. (CZ)
Šuman, J. (CZ)
Strnad, Hynek (UMG-J) RID
Ruml, T. (CZ)
Pačes, Václav (UMG-J) RID
Kotrba, P. (CZ)Source Title Research in Microbiology. - : Elsevier - ISSN 0923-2508
Roč. 164, č. 10 (2013), s. 1009-1018Number of pages 10 s. Language eng - English Country FR - France Keywords Achromobacter xylosoxidans ; metal tolerance ; lead ; cadmium ; zinc ; P1-type ATPase ; MerR family ; transcriptional control Subject RIV EB - Genetics ; Molecular Biology R&D Projects 1M0520 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Institutional support UMG-J - RVO:68378050 UT WOS 000329379400005 DOI 10.1016/j.resmic.2013.10.002 Annotation The cluster of pbtTFYRABC genes is carried by plasmid pA81. Its elimination from Achromobacter xylosoxidans A8 resulted in increased sensitivity towards Pb(2+) and Cd(2+). Predicted pbtTRABC products share strong similarities with Pb(2+) uptake transporter PbrT, transcriptional regulator PbrR, metal efflux P1-ATPases PbrA and CadA, undecaprenyl pyrophosphatase PbrB and its signal peptidase PbrC from Cupriavidus metallidurans CH34. Expression of pbtABC or pbtA in a metal-sensitive Escherichia coli GG48 rendered the strain Pb(2+)-, Cd(2+)- and Zn(2+)-tolerant and caused decreased accumulation of the metal ions. Accumulation of Pb(2+), but not of Cd(2+) or Zn(2+), was promoted in E. coli expressing pbtT. Additional genes of the pbt cluster are pbtF and pbtY, which encode the cation diffusion facilitator (CDF)-like transporter and a putative fatty acid hydroxylase of unknown function, respectively. Expression of pbtF did not confer increased metal tolerance upon E. coli GG48, although the protein showed measurable Pb(2+)-efflux activity. Unlike the pbtT promoter, promoters of pbtABC, pbtF and pbtY contain features characteristic of promoters controlled by metal-responsive transcriptional regulators of the MerR family. Upregulation of pbtABC, pbtF and pbtY upon Pb(2+), Cd(2+) and Zn(2+) exposure was confirmed in wild-type Achromobacter xylosoxidans A8. Gel shift assays proved binding of purified PbtR to the respective promoters. Copyright 2013 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2014
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