title: Transcriptional and functional studies of a Cd(II)/Pb(II)-responsive transcriptional regulator(CmtR) from Acidithiobacillus ferrooxidans ATCC 23270.
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, and Cys121. The results reported here strongly suggest that the high resistance of the extremophilic A. ferrooxidans to Cd including the Cd(II)/Pb(II)-responsive transcriptional regulator.
authors.affiliation: College of Environmental Science and Engineering, Donghua University, Shanghai, People's Republic of China. Zhengchunli1979@163.com
authors.forename: Chunli
authors.initials: C
authors.lastname: Zheng
authors.forename: Yanjun
authors.initials: Y
authors.lastname: Li
authors.forename: Li
authors.initials: L
authors.lastname: Nie
authors.forename: Lin
authors.initials: L
authors.lastname: Qian
authors.forename: Lu
authors.initials: L
authors.lastname: Cai
authors.forename: Jianshe
authors.initials: J
authors.lastname: Liu
mesh_terms: D042763:Acidithiobacillus / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D002104:Cadmium / Q000378:metabolism* / Q000633:toxicity; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D019295:Computational Biology; D024881:Drug Resistance, Bacterial; D004926:Escherichia coli / Q000235:genetics; D020869:Gene Expression Profiling; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D007854:Lead / Q000378:metabolism* / Q000633:toxicity; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D014157:Transcription Factors; D002104:Cadmium; D007854:Lead