title: A copper-responsive gene cluster is required for copper homeostasis and contributes to oxidative resistance in Deinococcus radiodurans R1.
abstract: Excess copper is toxic to organisms, and therefore, copper homeostasis is important for the limitation of its cellular levels. However, copper homeostasis has not been studied to date in the bacteria Deinococcus radiodurans R1, which exhibits extreme resistance to various environmental stresses. We have identified a copper-responsive gene cluster that encodes CopA, which is a copper-transporting P1-type ATPase, CopZ, which is a copper metallochaperone, and CsoR, which is a copper-sensing repressor. Copper induces the transcription of genes in this cluster. Mutants lacking copA exhibited reduced copper resistance and the overaccumulation of copper compared with the wild-type strain. Additionally, both in the absence and presence of copper, the copZ mutation increased the expression of copA and led to the accumulation of lower levels of copper compared with the wild type. The bioinformatic analysis showed that CsoR in D. radiodurans R1 shares high sequence similarity and identity with the CsoR of Mycobacterium tuberculosis and Bacillus subtilis. We also demonstrated through DNase I footprinting and electrophoretic mobility shift assays that CsoR binds to the promoter of the cluster and that copper ions eliminate this interaction. This implies that CsoR is the repressor of this cluster and that CopA, CopZ and CsoR participate in the regulation of copper homeostasis. Our data also indicate that after treatment with H2O2 and cumene hydroperoxide, the viability of the copA mutants was significantly reduced. This suggests that copper homeostasis plays an important role in oxidative resistance in D. radiodurans R1.
authors.affiliation: China University of Mining and Technology (Beijing), Beijing 100083, China. zhangyuxiu@cumtb.edu.cn.
authors.forename: Zhongchao
authors.initials: Z
authors.lastname: Zhao
authors.forename: Zhengfu
authors.initials: Z
authors.lastname: Zhou
authors.forename: Liang
authors.initials: L
authors.lastname: Li
authors.forename: Xianyi
authors.initials: X
authors.lastname: Xian
authors.forename: Xiubin
authors.initials: X
authors.lastname: Ke
authors.forename: Ming
authors.initials: M
authors.lastname: Chen
authors.forename: Yuxiu
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D003300:Copper / Q000378:metabolism*; D034301:Deinococcus / Q000235:genetics* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D023061:Gene Order; D006706:Homeostasis; D008969:Molecular Sequence Data; D005810:Multigene Family; D009154:Mutation; D018384:Oxidative Stress / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D016415:Sequence Alignment; D015533:Transcriptional Activation
chemical_list: D001426:Bacterial Proteins; D003300:Copper