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Applied and Environmental Microbiology, September 2003, p. 5104-5114, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5104-5114.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Sequence Diversity and Functional Conservation of the Origin of Replication in Lactococcal Prolate Phages

Jasna Rakonjac,1* Lawrence J. H. Ward,2 Anja H. Schiemann,1 Paul P. Gardner,3 Mark W. Lubbers,2 and Paul W. O'Toole1,{dagger}

Institute of Molecular BioSciences,1 Allan Wilson Centre for Molecular Ecology and Evolution, Massey University,3 Fonterra Research Centre, Palmerston North, New Zealand2

Received 14 October 2002/ Accepted 17 June 2003

Prolate or c2-like phages are a large homologous group of viruses that infect the bacterium Lactococcus lactis. In a collection of 122 prolate phages, three distinct, non-cross-hybridizing groups of origins of DNA replication were found. The nonconserved sequence was confined to the template for an untranslated transcript, PE1-T, 300 to 400 nucleotides in length, while the flanking sequences were conserved. All three origin types, despite the low sequence homology, have the same functional characteristics: they express abundant PE1-T transcripts and can function as origins of plasmid replication in the absence of phage proteins. Using chimeric constructs, we showed that hybrids of two nonhomologous origin sequences failed to function as replication origins, suggesting that preservation of a particular secondary structure of the PE1-T transcript is required for replication. This is the first systematic survey of the sequence and function of origins of replication in a group of lactococcal phages.

* Corresponding author. Mailing address: Institute of Molecular BioSciences, Massey University, Private Bag 11-222, Palmerston North, New Zealand. Phone: 64 6 350 5134. Fax: 64 6 350 5688. E-mail: J.Rakonjac{at}massey.ac.nz.

{dagger} Present address: Department of Microbiology, University College Cork, Cork, Ireland.

Applied and Environmental Microbiology, September 2003, p. 5104-5114, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5104-5114.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Rakonjac, J., O'Toole, P. W., Lubbers, M. (2005). Isolation of Lactococcal Prolate Phage-Phage Recombinants by an Enrichment Strategy Reveals Two Novel Host Range Determinants. J. Bacteriol. 187: 3110-3121 [Abstract] [Full Text]  
  • Schiemann, A. H., Rakonjac, J., Callanan, M., Gordon, J., Polzin, K., Lubbers, M. W., O'Toole, P. W. (2004). Essentiality of the Early Transcript in the Replication Origin of the Lactococcal Prolate Phage c2. J. Bacteriol. 186: 8010-8017 [Abstract] [Full Text]  


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