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Journal of Bacteriology, February 2005, p. 1044-1054, Vol. 187, No. 3
0021-9193/05/$08.00+0     doi:10.1128/JB.187.3.1044-1054.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Characterization of the Pheromone Response of the Enterococcus faecalis Conjugative Plasmid pCF10: Complete Sequence and Comparative Analysis of the Transcriptional and Phenotypic Responses of pCF10-Containing Cells to Pheromone Induction{dagger}

Helmut Hirt,1,2* Dawn A. Manias,1 Edward M. Bryan,1,{ddagger} Joanna R. Klein,1,§ Jesper K. Marklund,1 Jack H. Staddon,1 Michael L. Paustian,3,¶ Vivek Kapur,3 and Gary M. Dunny1

Department of Microbiology, Medical School, University of Minnesota, Minneapolis,1 Biomedical Genomics Center and Departments of Microbiology and Veterinary Pathobiology, University of Minnesota, St. Paul, Minnesota,3 Division of Biology, Kansas State University, Manhattan, Kansas2

Received 30 August 2004/ Accepted 28 October 2004

The sex pheromone plasmids in Enterococcus faecalis are one of the most efficient conjugative plasmid transfer systems known in bacteria. Plasmid transfer rates can reach or exceed 10–1 transconjugants per donor in vivo and under laboratory conditions. We report the completion of the DNA sequence of plasmid pCF10 and the analysis of the transcription profile of plasmid genes, relative to conjugative transfer ability following pheromone induction. These experiments employed a mini-microarray containing all 57 open reading frames of pCF10 and a set of selected chromosomal genes. A clear peak of transcription activity was observed 30 to 60 min after pheromone addition, with transcription subsiding 2 h after pheromone induction. The transcript activity correlated with the ability of donor cells to transfer pCF10 to recipient cells. Remarkably, aggregation substance (Asc10, encoded by the prgB gene) was present on the cell surface for a long period of time after pheromone-induced transcription of prgB and plasmid transfer ability had ceased. This observation could have relevance for the virulence of E. faecalis.

* Corresponding author. Mailing address: Division of Biology, Kansas State University, 232 Ackert Hall, Manhattan, KS 66506. Phone: (785) 532-2816. Fax: (785) 532-6653. E-mail: hhirt{at}ksu.edu.

{dagger} Contribution no. 05-68-J from the Kansas Agricultural Experiment Station.

{ddagger} Present address: Virologic, Inc., San Francisco, CA 94030.

§ Present address: Northwestern College, St. Paul, MN 55113.

Present address: National Veterinary Services Laboratories, Ames, IA 50010.

Journal of Bacteriology, February 2005, p. 1044-1054, Vol. 187, No. 3
0021-9193/05/$08.00+0     doi:10.1128/JB.187.3.1044-1054.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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