CodY Activates Transcription of a Small RNA in Bacillus subtilis▿ †
| Autor: | Rajani K. Gudipati, Heike Preis, Rita A. Eckart, Nadja Heidrich, Sabine Brantl |
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| Jazyk: | angličtina |
| Rok vydání: | 2009 |
| Předmět: |
DNA
Bacterial Small RNA Transcription Genetic Molecular Sequence Data DNA Footprinting DNA footprinting Bacillus subtilis Microbiology Bacterial Proteins Transcription (biology) Genes Reporter Gene Regulation RNA Small Interfering Promoter Regions Genetic Molecular Biology Gene Regulation of gene expression Genetics biology Base Sequence Gene Expression Profiling RNA Gene Expression Regulation Bacterial biology.organism_classification Blotting Northern beta-Galactosidase DNA binding site Trans-Activators Nucleic Acid Conformation Guanosine Triphosphate Amino Acids Branched-Chain Protein Binding |
| Popis: | Regulatory small RNAs (sRNAs) in bacterial genomes have become a focus of research over the past 8 years. Whereas more than 100 such sRNAs have been found in Escherichia coli , relatively little is known about sRNAs in gram-positive bacteria. Using a computational approach, we identified two sRNAs in intergenic regions of the Bacillus subtilis genome, SR1 and SR2 (renamed BsrF). Recently, we demonstrated that SR1 inhibits the translation initiation of the transcriptional activator AhrC. Here, we describe detection of BsrF, its expression profile, and its regulation by CodY. Furthermore, we mapped the secondary structure of BsrF. BsrF is expressed in complex and minimal media in all growth phases in B. subtilis and, with a similar expression profile, also in Bacillus amyloliquefaciens . Neither overexpression nor deletion of bsrF affected the growth of B. subtilis . BsrF was found to be long-lived in complex and minimal media. Analysis of 13 putative transcription factor binding sites upstream of bsrF revealed only an effect for CodY. Here, we showed by using Northern blotting, lacZ reporter gene fusions, in vitro transcription, and DNase I footprinting that the transcription of bsrF is activated by CodY in the presence of branched-chain amino acids and GTP. Furthermore, BsrF transcription was increased 1.5- to 2-fold by glucose in the presence of branched-chain amino acids, and this increase was independent of the known glucose-dependent regulators. BsrF is the second target for which transcriptional activation by CodY has been discovered. |
| Databáze: | OpenAIRE |
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