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Antimicrobial Agents and Chemotherapy, September 2003, p. 2868-2874, Vol. 47, No. 9
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.9.2868-2874.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Microcin C51 Plasmid Genes: Possible Source of Horizontal Gene Transfer

Dmitri E. Fomenko,1 Anastazia Z. Metlitskaya,1 Jean Péduzzi,2 Christophe Goulard,2 Genrikh S. Katrukha,3 Leonid V. Gening,1 Sylvie Rebuffat,2* and Inessa A. Khmel1

Institute of Molecular Genetics, Russian Academy of Sciences,1 Institute of New Antibiotics, Moscow, Russia,3 Department of Regulations, Development and Molecular Diversity, USM 502, National Museum of Natural History, UMR 5154 CNRS, 75231 Paris Cedex 05, France2

Received 16 December 2002/ Returned for modification 4 April 2003/ Accepted 5 June 2003

Microcin C51 (MccC51) is an antimicrobial nucleotide-heptapeptide produced by a natural Escherichia coli strain. A 5.7-kb fragment of the pC51 plasmid carrying the genes involved in MccC51 production, secretion, and self-immunity was sequenced, and the genes were characterized. The sequence of the MccC51 gene cluster is highly similar to that of the MccC7 gene. Recombinant plasmids carrying different combinations of the mcc genes involved in the MccC51 production or immunity were constructed to characterize their functional roles. The mccA, mccB, mccD, and mccE genes are involved in MccC51 production, while the mccC and mccE genes are responsible for immunity to MccC51. The mcc gene cluster is flanked by 44-bp direct repeats. Amino acid sequence comparisons allowed us to propose functions for each Mcc polypeptide in MccC51 biosynthesis. Plasmid pUHN containing the cloned mccA, mccB, mccC, and mccE genes, but lacking mccD, directed the synthesis of MccC51p, a substance chemically related to MccC51. MccC51p exhibited weak antibiotic activity against E. coli and was toxic to the producing cells. The immunity to exogenous MccC51 determined by the mccC and mccE genes did not overcome the toxic action of MccC51p on the producing cells. The G+C content of the MccC51 operon, markedly lower than that of the E. coli genome, and the presence of direct repeats suggest the possibility of horizontal transfer of this gene cluster.

* Corresponding author. Mailing address: Laboratoire de Chimie et Biochimie des Substances naturelles; Muséum National d'Histoire Naturelle, 63 rue Buffon, 75231 Paris Cedex 05, France. Phone: 33 1 40 79 31 18. Fax: 33 1 40 79 31 35. E-mail: rebuffat{at}mnhn.fr.

Antimicrobial Agents and Chemotherapy, September 2003, p. 2868-2874, Vol. 47, No. 9
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.9.2868-2874.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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