Transcriptional and functional studies of a Cd(II)/Pb(II)-responsive transcriptional regulator(CmtR) from Acidithiobacillus ferrooxidans ATCC 23270.

Current Microbiology
Chunli ZhengJianshe Liu

Abstract

The acidophilic Acidithiobacillus ferrooxidans can resist exceptionally high cadmium (Cd) concentrations. This property is important for its use in biomining processes, where Cd and other metal levels range usually between 15 and 100 mM. To learn about the mechanisms that allow A. ferrooxidans cells to survive in this environment, a bioinformatic search of its genome showed the presence of that a Cd(II)/Pb(II)-responsive transcriptional regulator (CmtR) was possibly related to Cd homeostasis. The expression of the CmtR was studied by real-time reverse transcriptase PCR using A. ferrooxidans cells adapted for growth in the presence of high concentrations of Cd. The putative A. ferrooxidans Cd resistance determinant was found to be upregulated when this bacterium was exposed to Cd in the range of 15-30 mM. The CmtR from A. ferrooxidans was cloned and expressed in Escherichia coli, the soluble protein was purified by one-step affinity chromatography to apparent homogeneity. UV-Vis spectroscopic measurements showed that the reconstruction CmtR was able to bind Cd(II) forming Cd(II)-CmtR complex in vitro. The sequence alignment and molecular modeling showed that the crucial residues for CmtR binding were likely to be Cys77, Cys112, ...Continue Reading

References

Feb 27, 1999·Molecular Microbiology·K R BrocklehurstA P Morby
Jun 28, 2003·FEMS Microbiology Reviews·Dietrich H Nies
Feb 27, 2004·The Journal of Biological Chemistry·Kate J Newberry, Richard G Brennan
Nov 23, 2005·Bioinformatics·Konstantin ArnoldTorsten Schwede

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Citations

Oct 15, 2013·Microbiology·Anna Jarosławiecka, Zofia Piotrowska-Seget
May 29, 2019·Journal of Structural Biology·Chandrasekaran PrabaharanSaravanamuthu Thiyagarajan

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Methods Mentioned

BETA
environmental stress
electrophoresis
PCR

Software Mentioned

Primer Premier
Swiss
ClustalX
Model
SwissModel

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