Published online 6 April 2005
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Experimental RNomics in Aquifex aeolicus: identification of small non-coding RNAs and the putative 6S RNA homolog
Philipps-Universität Marburg, Institut für Pharmazeutische Chemie Marbacher Weg 6, D-35037 Marburg, Germany 1Institute for Experimental Pathology, ZMBE, University of Münster Von-Esmarch-Strasse 56, D-48149 Münster, Germany 2Institute for Molecular Biology, Division of Functional Genomics and RNomics, Innsbruck Medical University A-6020 Innsbruck, Austria
*To whom correspondence should be addressed. Tel: +49 6421 28 25827; Fax: +49 6421 28 25854; Email: roland.hartmann{at}staff.uni-marburg.de
Received December 15, 2004. Revised March 16, 2005. Accepted March 16, 2005.
By an experimental RNomics approach, we have generated a cDNA library from small RNAs expressed from the genome of the hyperthermophilic bacterium Aquifex aeolicus. The library included RNAs that were antisense to mRNAs and tRNAs as well as RNAs encoded in intergenic regions. Substantial steady-state levels in A.aeolicus cells were confirmed for several of the cloned RNAs by northern blot analysis. The most abundant intergenic RNA of the library was identified as the 6S RNA homolog of A.aeolicus. Although shorter in size (150 nt) than its 
-proteobacterial homologs (
185 nt), it is predicted to have the most stable structure among known 6S RNAs. As in the -proteobacteria, the A.aeolicus 6S RNA gene (ssrS) is located immediately upstream of the ygfA gene encoding a widely conserved 5-formyltetrahydrofolate cyclo-ligase. We identifed novel 6S RNA candidates within the -proteobacteria but were unable to identify reasonable 6S RNA candidates in other bacterial branches, utilizing mfold analyses of the region immediately upstream of ygfA combined with 6S RNA blastn searches. By RACE experiments, we mapped the major transcription initiation site of A.aeolicus 6S RNA primary transcripts, located within the pheT gene preceding ygfA, as well as three processing sites.
Present address: Jens Minnerup, Universitätsklinikum Münster, Klinik und Poliklinik für Neurologie, Albert-Schweitzer-Strasse 33, D-48149 Münster, Germany
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