Characterization of chromosomal homologs of the plasmid-borne copper resistance operon of Pseudomonas syringae.
Copper-resistant and copper-sensitive strains of Pseudomonas syringae, as well as many other pseudomonads, contain chromosomal DNA homologous to the plasmid-borne copper resistance operon (copABCD). cop homologs were cloned from the chromosome of P. syringae pv. tomato PT12.2, which had an elevated level of resistance to copper compared with typical copper-sensitive strains of other P. syringae pathovars and showed an unusually high frequency of spontaneous mutation to high levels of copper resistance. Two chromosomal cop homolog regions were cloned. Homolog 1 hybridized with copA and copB, and homolog 2 hybridized with copA, copB, copC, and the copper-responsive regulatory genes copRS. Homolog 1 had no detectable function when transferred to a copper-sensitive strain of P. syringae. However, homolog 2 conferred the low level of copper resistance observed with PT12.2 and produced proteins related to CopA and CopC. In addition, homolog 2 conferred a high frequency of mutation to full copper resistance. In a spontaneously mutated derivative of the cloned homolog 2 (pCOPH2R) that conferred copper resistance, an increased level of CopA was observed. pCOPH2R also supported a higher level of transcriptional activity of the cop promoter that was fused to lacZ and provided in trans (pCOP38), suggesting that the spontaneous mutation was regulatory, probably involving the copRS homologs. Homolog 2 was similar but not identical to the plasmid-borne cop operon, and it did not complement site-specific mutations in cop genes.
Lim
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CK
Department of Plant Pathology, University of California, Riverside 92521-0122.
Cooksey
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DA
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D015139:Blotting, Southern; D027682:Cation Transport Proteins; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D003300:Copper / Q000633:toxicity*; D019327:Copper Sulfate; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004352:Drug Resistance, Microbial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005809:Genes, Regulator; D015698:Genomic Library; D008826:Microbial Sensitivity Tests; D009154:Mutation; D009876:Operon; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000187:drug effects; D011549:Pseudomonas / Q000187:drug effects / Q000235:genetics*; D011815:R Factors; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D027682:Cation Transport Proteins; C071263:CopA protein, Bacteria; D004269:DNA, Bacterial; D003300:Copper; D019327:Copper Sulfate