Original Paper

Antonie van Leeuwenhoek

, Volume 96, Issue 2, pp 161-170

First online:

ArsR arsenic-resistance regulatory protein from Cupriavidus metallidurans CH34

  • Yian-Biao ZhangAffiliated withBiology Department, Brookhaven National Laboratory
  • , Sébastien MonchyAffiliated withBiology Department, Brookhaven National LaboratoryLaboratory for Microbiology, Center of Studies for Nuclear Energy, SCK.CEN
  • , Bill GreenbergAffiliated withBiology Department, Brookhaven National Laboratory
  • , Max MergeayAffiliated withLaboratory for Microbiology, Center of Studies for Nuclear Energy, SCK.CEN
  • , Oleg GangAffiliated withCenter for Functional Nanomaterials, Brookhaven National Laboratory
  • , Safiyh TaghaviAffiliated withBiology Department, Brookhaven National Laboratory
  • , Daniel van der LelieAffiliated withBiology Department, Brookhaven National Laboratory Email author 

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The Cupriavidus metallidurans CH34 arsR gene, which is part of the arsRIC 2 BC 1 HP operon, and its putative arsenic-resistance regulatory protein were identified and characterized. The arsenic-induced transcriptome of C. metallidurans CH34 showed that the genes most upregulated in the presence of arsenate were all located within the ars operon, with none of the other numerous heavy metal resistance systems present in CH34 being induced. A transcriptional fusion between the luxCDABE operon and the arsR promoter/operator (P/O) region was used to confirm the in vivo induction of the ars operon by arsenite and arsenate. The arsR gene was cloned into expression vectors allowing for the overexpression of the ArsR protein as either his-tagged or untagged protein. The ability of the purified ArsR proteins to bind to the ars P/O region was analyzed in vitro by gel mobility shift assays. ArsR showed an affinity almost exclusively to its own ars P/O region. Dissociation of ArsR and its P/O region was metal dependent, and based on decreasing degrees of dissociation three groups of heavy metals could be distinguished: As(III), Bi(III), Co(II), Cu(II), Ni(II); Cd(II); Pb(II) and Zn(II), while no dissociation was observed in the presence of As(V).


Cupriavidus metallidurans CH34 Arsenic resistance ArsR Gene expression Metal binding