Bacillus subtilis RNase Y activity in vivo analysed by tiling microarrays

PLoS One. 2013;8(1):e54062. doi: 10.1371/journal.pone.0054062. Epub 2013 Jan 10.

Abstract

RNase Y is a key endoribonuclease affecting global mRNA stability in Bacillus subtilis. Its characterization provided the first evidence that endonucleolytic cleavage plays a major role in the mRNA metabolism of this organism. RNase Y shares important functional features with the RNA decay initiating RNase E from Escherichia coli, notably a similar cleavage specificity and a preference for 5' monophosphorylated substrates. We used high-resolution tiling arrays to analyze the effect of RNase Y depletion on RNA abundance covering the entire genome. The data confirm that this endoribonuclease plays a key role in initiating the decay of a large number of mRNAs as well as non coding RNAs. The downstream cleavage products are likely to be degraded by the 5' exonucleolytic activity of RNases J1/J2 as we show for a specific case. Comparison of the data with that of two other recent studies revealed very significant differences. About two thirds of the mRNAs upregulated following RNase Y depletion were different when compared to either one of these studies and only about 10% were in common in all three studies. This highlights that experimental conditions and data analysis play an important role in identifying RNase Y substrates by global transcriptional profiling. Our data confirmed already known RNase Y substrates and due to the precision and reproducibility of the profiles allow an exceptionally detailed view of the turnover of hundreds of new RNA substrates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus subtilis / enzymology
  • Bacillus subtilis / genetics
  • Bacillus subtilis / metabolism*
  • Endoribonucleases / metabolism
  • Escherichia coli / enzymology
  • Gene Expression Regulation, Bacterial
  • Genome, Bacterial
  • RNA Stability / genetics*
  • RNA, Messenger* / genetics
  • RNA, Messenger* / metabolism
  • RNA, Untranslated / genetics
  • RNA, Untranslated / metabolism
  • Ribonucleases* / genetics
  • Ribonucleases* / metabolism

Substances

  • RNA, Messenger
  • RNA, Untranslated
  • Endoribonucleases
  • Ribonucleases
  • ribonuclease E

Grants and funding

This work was supported by funds from the CNRS (UPR 9073), Univ Paris Diderot, Sorbonne Paris Cité and the Agence Nationale de la Recherche (http://www.agence-nationale-recherche.fr/) ANR grant RNAJAY. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.