Comparative transcriptome and proteome analyses of Bacillus subtilis 6S-1 and 6S-2 RNAs deletion strainsстатья
Статья опубликована в высокорейтинговом журнале
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Дата последнего поиска статьи во внешних источниках: 16 февраля 2016 г.
Аннотация: 6S RNAs are small non-coding regulatory RNAs found in many bacterial species that control gene expression on the transcriptional level by direct binding to RNA polymerase (RNAP) holoenzymes. Most bacteria express a single 6S RNA, while several species, mostly Firmicutes, encode two different 6S RNA homologs. Recent in vitro investigations performed for 6S-1 and 6S-2 RNAs from Bacillus subtilis provided evidence that both RNAs have canonical 6S RNA features and inhibit transcription by interaction with RNAP holoenzyme. Nevertheless, the functional role of each 6S RNA, having different expression profiles during cell growth, was unknown.
To analyze the cellular roles of B. subtilis 6S RNAs in vivo we constructed derivative strains of B. subtilis PY79 that carry deletions of the 6S-1 and 6S-2 RNA genes. Differential transcriptome analysis (RNAseq) of total RNA extracted from the phase of cell growth, when both 6S RNAs are actively expressed, revealed hundreds of genes, whose transcriptional activity significantly changed in absence of 6S-1 and/or 6S-2 RNAs. Two-dimensional differential gel electrophoresis of total protein extracts followed by MALDI mass-spectrometry allowed identification of a set of proteins that were increasingly particularly during exponential growth and preferentially in mutant strains lacking 6S-2 RNA. Several of these proteins are involved in metabolism and stress responses.