pestis microtus strain 201 was constructed in the present work. Microarray expression analysis disclosed a set of 154 Zur-dependent genes of Y. pestis upon exposure to zinc rich condition, and the microarray data was validated by real-time RT-PCR. Further biochemical assays, including LacZ reporter fusion, EMSA, DNase I footprinting, and primer extension, revealed that Zur as a repressor directly controlled the transcription
of znuA, znuCB and ykgM-RpmJ2 in Y. pestis by employing a conserved mechanism of Zur-promoter DNA association as observed in γ-Proteobacteria. It was thought that Zur contributed to zinc homeostasis in Y. pestis through transcriptional repression of the high-affinity zinc uptake system ZnuACB and two alternative ribosomal proteins YkgM https://www.selleckchem.com/products/OSI027.html and RpmJ2. Acknowledgements Financial supports BTSA1 mouse came from the National Natural Science Foundation of China for Distinguished Young Scholars (No. 30525025), the National Natural Science Foundation of China (No. 30771179), and the National Key Program for Infectious Disease of China (2009ZX10004-103 and 2008ZX10004-009). Electronic supplementary material Additional file 1: Colony counting of WT and Δzur upon exposure to 5 mM Zn. We performed colony counting of WT and Δzur upon exposure to
5 mM Zn for 30 min. The treatment with Zn had no toxic effect on both WT and Δzur. (DOC 40 KB) Additional file 2: Oligonucleiotide primers used in this study. (DOC 104 KB) Additional file 3: Zur-regulated genes grouped by functional classification according to Y. pestis CO92 genome annotation. Gene expression in Δzur was compared with that in the WT strain under Zn2+ rich (5 mM) condition. The Zur-regulated genes were divided into various functional categories. The numbers of up- and down-regulated genes were represented for Protein kinase N1 each functional group. (DOC 24 KB) Additional file 4: A complete list of Zur-regulated genes. (DOC 290 KB) Additional file 5: Comparison of transcription measurements by microarray and real-time PCR assays. The relative transcriptional levels for
17 genes selected from Supplementary Table S1 were determined by real-time RT-PCR. The log2 values were plotted against the microarray data log2 values. The correlation coefficient (R2) for comparison of the two datasets is 0.796. (DOC 174 KB) References 1. Hantke K: Bacterial zinc uptake and regulators. Curr Opin Microbiol 2005,8(2):196–202.CrossRefPubMed 2. Hantke K: Bacterial zinc transporters and regulators. Biometals 2001,14(3–4):239–249.CrossRefPubMed 3. Nies DH: Efflux-mediated heavy metal resistance in prokaryotes. FEMS Microbiol Rev 2003,27(2–3):313–339.CrossRefPubMed 4. Moore CM, Gaballa A, Hui M, Ye RW, Helmann JD: Genetic and physiological responses of Bacillus subtilis to metal ion stress. Mol Microbiol 2005,57(1):27–40.CrossRefPubMed 5. Perron K, Caille O, Rossier C, Van Delden C, Dumas JL, Kohler T: CzcR-CzcS, a two-component system involved in heavy metal and carbapenem resistance in Pseudomonas aeruginosa.