Abstract
Chromate-hypersensitive mutants of the Pseudomonas aeruginosa PAO1 strain were isolated using transposon-insertion mutagenesis. Comparison of the nucleotide sequences of the regions interrupted in the mutants with the PAO1 genome revealed that the genes affected in three mutant strains were oprE (ORF PA0291), rmlA (ORF PA5163), and ftsK (ORF PA2615), respectively. A relationship of these genes with chromate tolerance has not been previously reported. No other phenotypic changes were observed in the oprE mutant but its resistance to chromate was not fully restored by expressing the ChrA protein, which extrudes chromate ions from the cytoplasm to the periplasmic space. These data suggest that OprE participates in the efflux of chromate from the periplasm to the outside. Increased susceptibility of the rmlA mutant to the metals cadmium and mercury and to the anion-superoxide generator paraquat suggests a protective role of LPS against chromate toxicity. A higher susceptibility of the ftsK mutant to compounds affecting DNA structure (ciprofloxacin, tellurite, mitomycin C) suggests a role of FtsK in the recombinational repair of DNA damage caused by chromate. In conclusion, the P. aeruginosa genome contains diverse genes related to its intrinsic resistance to chromate. Systems pertaining to the outer membrane (OprE), the cell wall (LPS), and the cytoplasm (FtsK) were identified in this work as involved in chromate protection mechanisms.
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Acknowledgments
We thank the Pseudomonas Genome Project, the Pseudomonas Genetic Stock Center, and the Pseudomonas aeruginosa Community Annotation Project for use of the updated database and for cosmid donation. We also thank the Transposon Mutant Collection (University of Washington) for sending mutant strains. This work was supported by grants from Coordinación de la Investigación Científica (UMSNH) and Consejo Nacional de Ciencia y Tecnología (CONACYT; 41712Q). SLR was supported by a postgraduate fellowship from CONACYT.
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Rivera, S.L., Vargas, E., Ramírez-Díaz, M.I. et al. Genes related to chromate resistance by Pseudomonas aeruginosa PAO1. Antonie van Leeuwenhoek 94, 299–305 (2008). https://doi.org/10.1007/s10482-008-9247-x
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DOI: https://doi.org/10.1007/s10482-008-9247-x