Willkomm Dagmar K, Minnerup Jens, Hüttenhofer Alexander, Hartmann Roland K
Philipps-Universität Marburg, Institut für Pharmazeutische Chemie Marbacher Weg 6, D-35037 Marburg, Germany.
Nucleic Acids Res. 2005 Apr 6;33(6):1949-60. doi: 10.1093/nar/gki334. Print 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 gamma-proteobacterial homologs (approximately 185 nt), it is predicted to have the most stable structure among known 6S RNAs. As in the gamma-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 gamma-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.
通过一种实验性的RNA组学方法，我们从嗜热细菌嗜泉古菌（Aquifex aeolicus）基因组表达的小RNA中构建了一个cDNA文库。该文库包含与mRNA和tRNA反义的RNA以及基因间区域编码的RNA。通过Northern印迹分析证实了几种克隆RNA在嗜泉古菌细胞中的大量稳态水平。文库中最丰富的基因间RNA被鉴定为嗜泉古菌的6S RNA同源物。尽管其大小（150 nt）比其γ-变形菌同源物（约185 nt）短，但预计它在已知的6S RNA中具有最稳定的结构。与γ-变形菌一样，嗜泉古菌6S RNA基因（ssrS）位于编码广泛保守的5-甲酰四氢叶酸环化连接酶的ygfA基因的紧邻上游。我们在γ-变形菌中鉴定出了新的6S RNA候选物，但利用ygfA紧邻上游区域的mfold分析结合6S RNA blastn搜索，在其他细菌分支中未能鉴定出合理的6S RNA候选物。通过RACE实验，我们绘制了嗜泉古菌6S RNA初级转录本的主要转录起始位点，该位点位于ygfA之前的pheT基因内，以及三个加工位点。
Zotero 插件
