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Applied and Environmental Microbiology, April 2009, p. 2484-2494, Vol. 75, No. 8
0099-2240/09/$08.00+0     doi:10.1128/AEM.02093-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Identification and Characterization of the Phage Gene sav, Involved in Sensitivity to the Lactococcal Abortive Infection Mechanism AbiV

Jakob Haaber,¹ Geneviève M. Rousseau,² Karin Hammer,¹ and Sylvain Moineau^2*

Center for Systems Microbiology, Department of Systems Biology, Technical University of Denmark, DK-2800 Kongens Lyngby, Denmark,¹ Département de Biochimie et de Microbiologie, Faculté des Sciences et de Génie, Groupe de Recherche en Écologie Buccale, Faculté de Médecine Dentaire, Félix d'Hérelle Reference Center for Bacterial Viruses, Université Laval, Québec, Canada G1V 0A6²

Received 10 September 2008/ Accepted 15 February 2009

Lactococcus lactis phage mutants that are insensitive to the recently characterized abortive infection mechanism AbiV were isolated and analyzed in an effort to elucidate factors involved in the sensitivity to AbiV. Whole-genome sequencing of the phage mutants p2.1 and p2.2 revealed mutations in an orf that is transcribed early, indicating that this orf was responsible for AbiV sensitivity. Sequencing of the homologous regions in the genomes of other AbiV-insensitive mutants derived from p2 and six other lactococcal wild-type phages revealed point mutations in the homologous orf sequences. The orf was named sav (for sensitivity to AbiV), and the encoded polypeptide was named SaV. The purification of a His-tagged SaV polypeptide by gel filtration suggested that the polypeptide formed a dimer in its native form. The overexpression of SaV in L. lactis and Escherichia coli led to a rapid toxic effect. Conserved, evolutionarily related regions in SaV polypeptides of different phage groups are likely to be responsible for the AbiV-sensitive phenotype and the toxicity.

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* Corresponding author. Mailing address: Groupe de Recherche en Écologie Buccale, Faculté de Médecine Dentaire, Université Laval, Quebec City, Quebec, Canada G1V 0A6. Phone: (418) 656-3712. Fax: (418) 656-2861. E-mail: Sylvain.Moineau{at}bcm.ulaval.ca

Published ahead of print on 6 March 2009.

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Applied and Environmental Microbiology, April 2009, p. 2484-2494, Vol. 75, No. 8
0099-2240/09/$08.00+0     doi:10.1128/AEM.02093-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

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