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Journal of Virology, May 1999, p. 3920-3929, Vol. 73, No. 5
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Cooperative Interaction of CI Protein Regulates Lysogeny of Lactobacillus casei by Bacteriophage A2

Pilar García,1,2 Victor Ladero,1 Juan C. Alonso,3 and Juan E. Suárez1,2,*

Area de Microbiología, Facultad de Medicina, Universidad de Oviedo, 33007 Oviedo,1 Instituto de Productos Lácteos de Asturias (CSIC), 33400 Villaviciosa,2 and Centro Nacional de Biotecnología, Campus de la Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid,3 Spain

Received 28 December 1998/Accepted 8 February 1999

The temperate bacteriophage A2 forms stable lysogens in Lactobacillus casei. The A2-encoded cI product (CI), which is responsible for maintaining the A2 prophage in the lysogenic state, has been purified. The CI protein, which is a monomer of 25.3 kDa in solution, specifically binds to a 153-bp DNA fragment that contains two divergent promoters, PL and PR. These promoters mediate transcription from cI and a putative cro, respectively. Three similar, although not identical, 20-bp inverted repeated DNA segments (operator sites O1, O2, and O3) were found in this segment. CI selectively interacts with O1, which is placed downstream from the transcription start point of the cro gene, and with O2 and O3, which overlap with the -35 region of the two promoters. Using a heterologous RNA polymerase, we have determined the transcription start points of PL and PR. CI exerts a negative effect on the in vitro transcription of PR by repositioning the RNA polymerase in a concentration-dependent manner. CI, when bound to O1 and O2, enhances the positioning of the RNA polymerase with the PL promoter. Our data indicate that the CI protein regulates the lytic and lysogenic pathways of the A2 phage.

* Corresponding author. Mailing address: Area de Microbiología, Facultad de Medicina, Universidad de Oviedo, Julián Clavería s. n., 33007 Oviedo, Spain. Phone: 34 985 103559. Fax: 34 985 103148. E-mail: jsuarez{at}sauron.quimica.uniovi.es.

Journal of Virology, May 1999, p. 3920-3929, Vol. 73, No. 5
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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