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Microbiology 155 (2009), 2994-3004; DOI  10.1099/mic.0.028035-0
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Microbiology 155 (2009), 2994-3004; DOI  10.1099/mic.0.028035-0
© 2009 Society for General Microbiology

Illumination stimulates cAMP receptor protein-dependent transcriptional activation from regulatory regions containing class I and class II promoter elements in Synechocystis sp. PCC 6803

Jennifer Hedger1,{dagger}, Peter C. Holmquist1, Kimberly A. Leigh2, Kumuda Saraff1, Christina Pomykal1 and Michael L. Summers1

1 California State University Northridge, Department of Biology, 18111 Nordhoff St, Northridge, CA 91330, USA
2 Amgen, Thousand Oaks, CA 91320, USA

The cAMP receptor protein (Crp) is a global transcriptional regulator that binds sequence-specific promoter elements when associated with cAMP. In the motile cyanobacterium Synechocystis sp. strain PCC 6803, intracellular cAMP increases when dark-adapted cells are illuminated. Previous work has established that Crp binds proposed Crp target sites upstream of slr1351 (murF), sll1874 (chlAII), sll1708 (narL), slr0442 and sll1268 in vitro, and that slr0442 is downregulated in a crp mutant during photoautotrophic growth. To identify additional Crp target genes in Synechocystis, 11 different Crp binding sites proposed during a previous computational survey were tested for in vitro sequence-specific binding and crp-dependent transcription. The results indicate that murF, chlAII and slr0442 can be added as ‘target genes of Sycrp1’ in Synechocystis. Promoter mapping of the targets revealed the same close association of RNA polymerase and Crp as that found in Escherichia coli class I and class II Crp-regulated promoters, thereby strongly suggesting similar mechanisms of transcriptional activation.

Correspondence
Michael L. Summers
michael.l.summers{at}csun.edu

Abbreviations: EMSA, electrophoretic mobility shift assay; MSA, multiple sequence alignment; RACE, rapid amplification of cDNA ends; RT-QPCR, reverse transcriptase-mediated quantitative PCR; RNAP, RNA polymerase

{dagger}Present address: C3 Jian, Inc., Inglewood, CA 90301, USA.

A supplementary table, listing primers used in this study, is available with the online version of this paper.






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