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Applied and Environmental Microbiology, December 2001, p. 5626-5633, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.12.5626-5633.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

An Activator of Transcription Regulates Phage TP901-1 Late Gene Expression

Lone Brøndsted,* Margit Pedersen, and Karin Hammer

Department of Microbiology, Technical University of Denmark, DK-2800 Lyngby, Denmark

Received 6 August 2001/Accepted 1 October 2001

A promoter active in the late phase of the lytic cycle of lactococcal bacteriophage TP901-1 has been identified. The promoter is tightly regulated and requires the product of the phage TP901-1 orf29 for activity. A deletion analysis of the late promoter region showed that a fragment as small as 99 bp contains both the promoter and the region necessary for activation by ORF29. The transcriptional start site of the promoter was identified by primer extension to position 13073 on the TP901-1 genome, thus located 87 bp downstream of orf29 in a 580-bp intergenic region between orf29 and orf30. Furthermore, the region located -85 to -61 bp upstream of the start site was shown to be necessary for promoter activity. During infection, the transcript arising from the late promoter is fully induced at 40 min postinfection, and our results suggest that a certain level of ORF29 must be reached in order to activate transcription of the promoter. Several lactococcal bacteriophages encode ORF29 homologous proteins, indicating that late transcription may be controlled by a similar mechanism in these phages. With the identification of this novel regulator, our results suggest that within the P335 group of lactococcal phages at least two regulatory systems controlling transcription in the late stage of infection exist.


* Corresponding author. Present address: Department of Dairy and Food Science, Royal Veterinary and Agricultural University of Denmark, Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark. Phone: 45 35 28 32 71. Fax: 45 35 28 32 14. E-mail: lobr{at}kvl.dk.


Applied and Environmental Microbiology, December 2001, p. 5626-5633, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.12.5626-5633.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.



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