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Molecular genetics of a chromosomal locus involved in copper tolerance in Escherichia coli K-12.
Mol Microbiol. 1995 Mar; 15(6):1127-37.MM

Abstract

The cutA locus, presumably involved in copper tolerance in Escherichia coli, was characterized by a mutation leading to copper sensitivity. Copper-accumulation measurements with radioactive 64Cu2+ showed increased uptake by cutA copper-sensitive mutant cells, and reduced uptake when the cutA mutation was complemented in trans. The locus was mapped using complementation of the cutA mutant to partial copper tolerance with wild-type chromosomal fragments. The 3.2 kb DNA region involved in cutA was sequenced and analysed, revealing three significant open reading frames, none of which had been previously published. The products of all three open reading frames were identified, when synthesized with the T7 phage promoter expression system, as polypeptides of about 50 kDa, 24 kDa, and 13 kDa, consistent with the sizes predicted from the DNA sequences. The 50 kDa and 24 kDa polypeptides were found in the bacterial inner membrane, and the 13 kDa polypeptide with the cytoplasmic fraction. In addition to being required for copper tolerance, cutA affects tolerance levels to zinc, nickel, cobalt and cadmium salts. Transcriptional fusions of cutA with the lux operon showed induction by copper, zinc, nickel, cobalt and, to a lesser extent, cadmium, manganese and silver salts.

Authors+Show Affiliations

Department of Genetics, University of Melbourne, Parkville, Victoria, Australia.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

7623666

Citation

Fong, S T., et al. "Molecular Genetics of a Chromosomal Locus Involved in Copper Tolerance in Escherichia Coli K-12." Molecular Microbiology, vol. 15, no. 6, 1995, pp. 1127-37.
Fong ST, Camakaris J, Lee BT. Molecular genetics of a chromosomal locus involved in copper tolerance in Escherichia coli K-12. Mol Microbiol. 1995;15(6):1127-37.
Fong, S. T., Camakaris, J., & Lee, B. T. (1995). Molecular genetics of a chromosomal locus involved in copper tolerance in Escherichia coli K-12. Molecular Microbiology, 15(6), 1127-37.
Fong ST, Camakaris J, Lee BT. Molecular Genetics of a Chromosomal Locus Involved in Copper Tolerance in Escherichia Coli K-12. Mol Microbiol. 1995;15(6):1127-37. PubMed PMID: 7623666.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular genetics of a chromosomal locus involved in copper tolerance in Escherichia coli K-12. AU - Fong,S T, AU - Camakaris,J, AU - Lee,B T, PY - 1995/3/1/pubmed PY - 1995/3/1/medline PY - 1995/3/1/entrez SP - 1127 EP - 37 JF - Molecular microbiology JO - Mol Microbiol VL - 15 IS - 6 N2 - The cutA locus, presumably involved in copper tolerance in Escherichia coli, was characterized by a mutation leading to copper sensitivity. Copper-accumulation measurements with radioactive 64Cu2+ showed increased uptake by cutA copper-sensitive mutant cells, and reduced uptake when the cutA mutation was complemented in trans. The locus was mapped using complementation of the cutA mutant to partial copper tolerance with wild-type chromosomal fragments. The 3.2 kb DNA region involved in cutA was sequenced and analysed, revealing three significant open reading frames, none of which had been previously published. The products of all three open reading frames were identified, when synthesized with the T7 phage promoter expression system, as polypeptides of about 50 kDa, 24 kDa, and 13 kDa, consistent with the sizes predicted from the DNA sequences. The 50 kDa and 24 kDa polypeptides were found in the bacterial inner membrane, and the 13 kDa polypeptide with the cytoplasmic fraction. In addition to being required for copper tolerance, cutA affects tolerance levels to zinc, nickel, cobalt and cadmium salts. Transcriptional fusions of cutA with the lux operon showed induction by copper, zinc, nickel, cobalt and, to a lesser extent, cadmium, manganese and silver salts. SN - 0950-382X UR - https://www.unboundmedicine.com/medline/citation/7623666/Molecular_genetics_of_a_chromosomal_locus_involved_in_copper_tolerance_in_Escherichia_coli_K_12_ L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&sid=nlm:pubmed&issn=0950-382X&date=1995&volume=15&issue=6&spage=1127 DB - PRIME DP - Unbound Medicine ER -