Jianqiang Liang, Mingzhe Zhang, Mingmei Lu, Zhefei Li, Xihui Shen, Minxia Chou, Gehong Wei. Functional Characterization of a csoR-cueA Divergon in Bradyrhizobium Liaoningense CCNWSX0360, Involved in Copper, Zinc and Cadmium Cotolerance

Functional Characterization of a csoR-cueA Divergon in Bradyrhizobium Liaoningense CCNWSX0360, Involved in Copper, Zinc and Cadmium Cotolerance.

Jianqiang Liang, Mingzhe Zhang, Mingmei Lu, Zhefei Li, Xihui Shen, Minxia Chou, Gehong Wei.

Scientific RepoRts

DOI: 10.1038/srep35155

Abstract: Random mutagenesis in a symbiotic nitrogen-fxing Bradyrhizobium liaoningense CCNWSX0360 (Bln0360) using Tn5 identifed fve copper (Cu) resistance-related genes. They were functionally sorted into three groups: transmembrane transport (cueA and tolC); oxidation (copA); and protection of the membrane barrier (lptE and ctpA). The gene cueA, together with the upstream csoR (Cu-sensitive operon repressor), constituted a csoR-cueA divergon which plays a crucial role in Cu homeostasis. Deletion of cueA decreased the Cu tolerance of cells, and complementation of this mutant restored comparable Cu resistance to that of the wild-type. Transcriptional and fusion expression analysis demonstrated that csoR-cueA divergon was up-regulated by both the monovalent Cu+ and divalent Zn2+/Cd2+, and negatively regulated by transcriptional repressor CsoR, via a bidirectional promoter. Deletion of csoR renders the cell hyper-resistant to Cu, Zn and Cd. Although predicted to encode a Cu transporting P-type ATPase (CueA), cueA also conferred resistance to zinc and cadmium; two putative N-MBDs (N-terminal metal binding domains) of CueA were required for the Cu/Zn/Cd tolerance. Moreover, cueA is needed for nodulation competitiveness of B. liaoningense in Cu rich conditions. Together, the results demonstrated a crucial role for the csoR-cueA divergon as a component of the multiple-metal resistance machinery in B. liaoningense.