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Journal of Bacteriology, November 2002, p. 6325-6332, Vol. 184, No. 22
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.22.6325-6332.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Characterization of the Two-Component Abortive Phage Infection Mechanism AbiT from Lactococcus lactis

Julie D. Bouchard, Eric Dion, Frédéric Bissonnette, and Sylvain Moineau*

Department of Biochemistry and Microbiology, Faculté des Sciences et de Génie, Groupe de Recherche en Écologie Buccale, Faculté de Médecine Dentaire, Université Laval, Québec, Canada G1K 7P4

Received 22 April 2002/ Accepted 23 July 2002

During the production of fermented dairy products, virulent bacteriophages infecting Lactococcus lactis can delay or stop the milk acidification process. A solution to this biological problem consists of introducing natural phage barriers into the strains used by the dairy industry. One such hurdle is called abortive infection (Abi) and causes premature cell death with no or little phage progeny. Here, we describe the isolation and characterization of a novel Abi mechanism encoded by plasmid pED1 from L. lactis. The system is composed of two constitutively cotranscribed genes encoding putative proteins of 127 and 213 amino acids, named AbiTi and AbiTii, respectively. Site-directed mutagenesis indicated that a hydrophobic region at the C-terminal extremity of AbiTi is essential to the antiphage phenotype. The AbiT system is effective against phages of the 936 and P335 species (efficiency of plaquing between 10-5 and 10-7) and causes a 20-fold reduction in the efficiency to form centers of infection as well as a 10- to 12-fold reduction in the burst size. Its efficacy could be improved by raising the plasmid copy number, but changing the intrinsic ratio of AbiTi and AbiTii did not greatly affect the antiphage activity. The monitoring of the intracellular phage infection process by DNA replication, gene expression, and electron microscopy as well as the study of phage mutants by genome mapping indicated that AbiT is likely to act at a later stage of the phage lytic cycle.

* Corresponding author. Mailing address: Groupe de recherche en écologie buccale (GREB), Faculté de médecine dentaire, Université Laval, Québec, Canada G1K 7P4. Phone: (418) 656-3712. Fax: (418) 656-2861. E-mail: Sylvain.Moineau{at}bcm.ulaval.ca.

Journal of Bacteriology, November 2002, p. 6325-6332, Vol. 184, No. 22
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.22.6325-6332.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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