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Journal of Bacteriology, June 2004, p. 3480-3491, Vol. 186, No. 11
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.11.3480-3491.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Bacteriophage Tuc2009 Encodes a Tail-Associated Cell Wall-Degrading Activity

John G. Kenny,1 Stephen McGrath,1 Gerald F. Fitzgerald,1,2 and Douwe van Sinderen1,2*

National Food Biotechnology Centre and Department of Microbiology,1 Alimentary Pharmabiotic Centre, Biosciences Institute, National University of Ireland, Cork, Ireland2

Received 1 December 2003/ Accepted 23 February 2004

Tuc2009 is a P335-type member of the tailed-phage supergroup Siphoviridae and was originally identified as a resident prophage of the gram-positive bacterium Lactococcus lactis UC509. A Tuc2009 gene designated tal2009 which is located within the morphogenic module was shown to specify a lytic activity within the 3' portion of its coding region. Comparative sequence analysis indicated that the cell wall-degrading part of Tal2009 is a member of the M37 protein family and that Tal2009 lacks a cell-binding domain, a finding supported by binding studies. Tal2009 appears to undergo self-mediated posttranslational processing in both L. lactis and Escherichia coli. Antibodies directed against a purified C-terminal portion of Tal2009 were used for immunoelectron microscopy, which showed that Tal2009 is located at the tail tip of Tuc2009. Antibody neutralization studies demonstrated that Tal2009-directed antibodies inhibited the ability of phage to mediate host lysis by more than 100-fold. These data indicate that tal2009 encodes a tail-associated lysin involved in localized cell wall degradation, thus allowing the Tuc2009 DNA injection machinery access to the membrane of its bacterial host.

* Corresponding author. Mailing address: National Food Biotechnology Centre and Department of Microbiology, National University of Ireland, Cork, Ireland. Phone: 353 21 490 1365. Fax: 353 21 490 3101. E-mail: d.vansinderen{at}ucc.ie.

Journal of Bacteriology, June 2004, p. 3480-3491, Vol. 186, No. 11
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.11.3480-3491.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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