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Nucleic Acids Research, 2002, Vol. 30, No. 7 1539-1546
© 2002 Oxford University Press

Rhodobacter sphaeroides LexA has dual activity: optimising and repressing recA gene transcription

Angels Tapias, Silvia Fernández1, Juan C. Alonso1 and Jordi Barbé*

Departamento de Genética y Microbiología, Universitat Autónoma de Barcelona, Bellaterra, 08193 Barcelona, Spain and 1Departamento de Biotecnología Microbiana, Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain

Transcription of the Rhodobacter sphaeroides recA promoter (PrecA) is induced upon DNA damage in a lexA-dependent manner. In vivo experiments demonstrate that LexA protein represses and might also activate transcription of PrecA. Purified R.sphaeroides LexA protein specifically binds the SOS boxes located within the PrecA region. In vitro transcription analysis, using Escherichia coli RNA polymerase (RNAP), indicated that the presence of LexA may stimulate and repress transcription of PrecA. EMSA and DNase I footprinting experiments show that LexA and RNAP can bind simultaneously to PrecA. At low LexA concentrations it enhances RNAP binding to PrecA, stimulates open complex formation and strand separation beyond the transcription start site. At high LexA concentrations, however, RNAP-promoted strand separation is not observed beyond the +5 region. LexA might repress transcription by interfering with the clearance process instead of blocking the access of RNAP to the promoter region. Based on these findings we propose that the R.sphaeroides LexA protein performs fine tuning of the SOS response, which might provide a physiological advantage by enhancing transcription of SOS genes and delaying full activation of the response.

* To whom correspondence should be addressed. Tel: +34 93 581 18 37; Fax: +34 93 581 23 87; Email: jordi.barbe{at}uab.es Correspondence may also be addressed to Juan C. Alonso. Tel: +34 91 585 45 46; Fax: +34 91 585 45 06; Email: jcalonso{at}cnb.uam.es Present address: Angels Tapias, Institute de Genetique et de Biologie Moleculaire et Cellulaire, IGBMC 1, Rue Laurent Fris, BP163, 67400 Illkirch, France The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors


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