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

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 m...

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Published in:	Microbiology (Society for General Microbiology) Vol. 155; no. 9; pp. 2994 - 3004
Main Authors:	Hedger, Jennifer, Holmquist, Peter C, Leigh, Kimberly A, Saraff, Kumuda, Pomykal, Christina, Summers, Michael L
Format:	Journal Article
Language:	English
Published:	Reading Soc General Microbiol 01-09-2009 Society for General Microbiology Microbiology Society
Subjects:	Bacterial Proteins - metabolism Bacteriology Base Sequence Biological and medical sciences Cell and Molecular Biology of Microbes Cyanobacteria DNA, Bacterial - genetics DNA, Bacterial - metabolism Environment Escherichia coli Fundamental and applied biological sciences. Psychology Gene Expression Regulation, Bacterial Genes, Reporter Genetics Luminescence Microbiology Molecular Sequence Data Peptide Synthases - biosynthesis Peptide Synthases - genetics Promoter Regions, Genetic Protein Binding Receptors, Cyclic AMP - metabolism Synechocystis Synechocystis - genetics Synechocystis - metabolism Transcription Initiation Site Transcriptional Activation Transcription Transcription promoter Light Cyanobacteria Cyclic AMP Bacteria Activation Synechocystis Protein
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Summary:	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 ( chlA II ), 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, chlA II 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 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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Present address: C3 Jian, Inc., Inglewood, CA 90301, USA.
ISSN:	1350-0872 1465-2080
DOI:	10.1099/mic.0.028035-0