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Journal of Bacteriology, May 2004, p. 2567-2575, Vol. 186, No. 9
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.9.2567-2575.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Identification of PimR as a Positive Regulator of Pimaricin Biosynthesis in Streptomyces natalensis

Nuria Antón,1 Marta V. Mendes,1 Juan F. Martín,1,2 and Jesús F. Aparicio1,2*

Institute of Biotechnology INBIOTEC, Parque Científico de León, 24006 León,1 Area of Microbiology, Faculty of Biology, University of León, 24071 León, Spain2

Received 4 December 2003/ Accepted 15 January 2004

Sequencing of the DNA region on the left fringe of the pimaricin gene cluster revealed the presence of a 3.6-kb gene, pimR, whose deduced product (1,198 amino acid residues) was found to have amino acid sequence homology with bacterial regulatory proteins. Database comparisons revealed that PimR represents the archetype of a new class of regulators, combining a Streptomyces antibiotic regulatory protein (SARP)-like N-terminal section with a C-terminal half homologous to guanylate cyclases and large ATP-binding regulators of the LuxR family. Gene replacement of pimR from Streptomyces natalensis chromosome results in a complete loss of pimaricin production, suggesting that PimR is a positive regulator of pimaricin biosynthesis. Gene expression analysis by reverse transcriptase PCR (RT-PCR) of the pimaricin gene cluster revealed that S. natalensis {Delta}PimR shows no expression at all of the cholesterol oxidase-encoding gene pimE, and very low level transcription of the remaining genes of the cluster except for the mutant pimR gene, thus demonstrating that this regulator activates the transcription of all the genes belonging to the pimaricin gene cluster but not its own transcription.

* Corresponding author. Mailing address: Institute of Biotechnology INBIOTEC, Parque Científico de León, Avda. del Real, no. 1, 24006 León, Spain. Phone: 34-987-210308. Fax: 34-987-210388. E-mail: degjfa{at}unileon.es.

Journal of Bacteriology, May 2004, p. 2567-2575, Vol. 186, No. 9
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.9.2567-2575.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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