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Volume 147, Issue 3

Characterization of a copper-transport operon, , from Free

Abstract

A copper-transport () operon was cloned from the oral bacterium JH1005. DNA sequencing showed that the operon contained three genes (, and ), which were flanked by a single promoter and a factor-independent terminator. encoded a small protein of 147 aa with a heavy-metal-binding motif (CXCXCXC) at the C-terminus. CopY shared extensive homology with other bacterial negative transcriptional regulators. encoded a 742 aa protein that shared extensive homology with P-type ATPases. encoded a 67 aa protein that also contained a heavy-metal-binding motif (CXXC) at the N-terminus. Northern blotting showed that a 32 kb transcript was produced by Cu-induced cells, suggesting that the genes were synthesized as a polycistronic message. The transcriptional start site of the operon was mapped and shown to lie within the inverted repeats of the promoter–operator region. wild-type cells were resistant to 800 μM Cu, whereas cells of a knock-out mutant were killed by 200 μM Cu. Complementation of the knock-out mutant with the operon restored Cu resistance to wild-type level. The wild-type and the mutant did not show any differences in susceptibility to other heavy metals, suggesting that the operon was specific for copper. By using a chloramphenicol acetyltransferase reporter gene fusion, the operon was shown to be negatively regulated by CopY and could be derepressed by Cu.

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  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): CAT, chloramphenicol acetyltransferase , copper resistance , copper transport , P-type ATPase, heavy metals , THA, Todd–Hewitt agar , THB, Todd–Hewitt broth and TYG, tryptone/yeast extract glucose broth
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2001-03-01
2023-03-21
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Characterization of a copper-transport operon, copYAZ, from Streptococcus mutans
Microbiology 147, 653 (2001); https://doi.org/10.1099/00221287-147-3-653
/content/journal/micro/10.1099/00221287-147-3-653
/content/journal/micro/10.1099/00221287-147-3-653
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