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J Gen Virol 87 (2006), 2423-2431; DOI 10.1099/vir.0.81875-0

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© 2006 Society for General Microbiology

Expression of phage P4 integrase is regulated negatively by both Int and Vis

D. Piazzolla{dagger}, S. Calì{ddagger}, E. Spoldi§, F. Forti, C. Sala, F. Magnoni, G. Dehò and D. Ghisotti

Dipartimento di Scienze Biomolecolari e Biotecnologie, Università degli Studi di Milano, Via Celoria 26, 20133 Milano, Italy

Correspondence
D. Ghisotti
Daniela.Ghisotti{at}unimi.it

Phage P4 int gene encodes the integrase responsible for phage integration into and excision from the Escherichia coli chromosome. Here, the data showing that P4 int expression is regulated in a complex manner at different levels are presented. First of all, the Pint promoter is regulated negatively by both Int and Vis, the P4 excisionase. The N-terminal portion of Int appears to be sufficient for such a negative autoregulation, suggesting that the Int N terminus is implicated in DNA binding. Second, full-length transcripts covering the entire int gene could be detected only upon P4 infection, whereas in P4 lysogens only short 5'-end covering transcripts were detectable. On the other hand, transcripts covering the 5'-end of int were also very abundant upon infection. It thus appears that premature transcription termination and/or mRNA degradation play a role in Int-negative regulation both on the basal prophage transcription and upon infection. Finally, comparison between PintlacZ transcriptional and translational fusions suggests that Vis regulates Int expression post-transcriptionally. The findings that Vis is also an RNA-binding protein and that Int may be translated from two different start codons have implications on possible regulation models of Int expression.

{dagger}Present address: Max F. Perutz Laboratories at the Vienna Biocenter, Department of Microbiology and Genetics, University of Vienna, 1030 Vienna, Austria.

{ddagger}Present address: Keryos, Via della Filanda 5, 20060 Gessate (MI), Italy.

§Present address: Molmed SpA, Via Olgettina 58, 20132 Milano, Italy.




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