Nucleic Acids Research, 2003, Vol. 31, No. 22 6435-6443
© 2003 Oxford University Press
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RNomics in Escherichia coli detects new sRNA species and indicates parallel transcriptional output in bacteria
1 Institute of Cell and Molecular Biology, Biomedical Center, Uppsala University, Box 596, 75124 Uppsala, Sweden, 2 Department of Molecular Genetics and Biotechnology, The Hebrew University-Hadassah Medical School, PO Box 12272, 91120 Jerusalem, Israel, 3 Institut für Experimentelle Pathologie/Molekulare Neurobiologie, Universität Münster, Von-Esmarch-Str. 56, D-48149 Münster, Germany, 4 Department of Medical Microbiology and Parasitology, School of Medical Sciences, 16150 Kubang Kerian, Kelantan, Malaysia and 5 Institut für Molekularbiologie, Abt. Funktionelle Genomik, Universität Innsbruck, Peter-Mayr-Str. 4b, 6020 Innsbruck, Austria
*To whom correspondence should be addressed Tel: +46 18 471 4579; Fax: +46 18 530 396; Email: joerg.vogel{at}icm.uu.se
Correspondence may also be addressed to Alexander Hüttenhofer. Tel: +43 512 507 3630; Fax: +43 512 507 9880; Email: alexander.huettenhofer{at}uibk.ac.at
Recent bioinformatics-aided searches have identified many new small RNAs (sRNAs) in the intergenic regions of the bacterium Escherichia coli. Here, a shot-gun cloning approach (RNomics) was used to generate cDNA libraries of small sized RNAs. Besides many of the known sRNAs, we found new species that were not predicted previously. The present work brings the number of sRNAs in E.coli to 62. Experimental transcription start site mapping showed that some sRNAs were encoded from independent genes, while others were processed from mRNA leaders or trailers, indicative of a parallel transcriptional output generating sRNAs co-expressed with mRNAs. Two of these RNAs (SroA and SroG) consist of known (THI and RFN) riboswitch elements. We also show that two recently identified sRNAs (RyeB and SraC/RyeA) interact, resulting in RNase III-dependent cleavage. To the best of our knowledge, this represents the first case of two non-coding RNAs interacting by a putative antisense mechanism. In addition, intracellular metabolic stabilities of sRNAs were determined, including ones from previous screens. The wide range of half-lives (<2 to >32 min) indicates that sRNAs cannot generally be assumed to be metabolically stable. The experimental characterization of sRNAs analyzed here suggests that the definition of an sRNA is more complex than previously assumed.
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