Three Essential Ribonucleases—RNase Y, J1, and III—Control the Abundance of a Majority of Bacillus subtilis mRNAs

摘要

Bacillus subtilis possesses three essential enzymes thought to be involved in mRNA decay to varying degrees, namely RNase Y, RNase J1, and RNase III. Using recently developed high-resolution tiling arrays, we examined the effect of depletion of each of these enzymes on RNA abundance over the whole genome. The data are consistent with a model in which the degradation of a significant number of transcripts is dependent on endonucleolytic cleavage by RNase Y, followed by degradation of the downstream fragment by the 5′–3′ exoribonuclease RNase J1. However, many full-size transcripts also accumulate under conditions of RNase J1 insufficiency, compatible with a model whereby RNase J1 degrades transcripts either directly from the 5′ end or very close to it. Although the abundance of a large number of transcripts was altered by depletion of RNase III, this appears to result primarily from indirect transcriptional effects. Lastly, RNase depletion led to the stabilization of many low-abundance potential regulatory RNAs, both in intergenic regions and in the antisense orientation to known transcripts. Author Summary RNA turnover is an important way of controlling gene expression. While the characterization of the pathways and enzymes for RNA degradation are well-advanced in Escherichia coli and yeast, studies in Gram-positive bacteria have lagged behind. This tiling array study shows that two essential enzymes, the single-strand specific endonuclease RNase Y and the 5′–3′ exoribonuclease RNase J1, play central roles in the degradation of mRNAs in Bacillus subtilis . The double-strand specific enzyme RNase III plays a more minor role in RNA turnover, but has significant indirect effects on transcription. Depleting cells of these key enzymes led to the stabilization of many potentially regulatory RNAs, which were otherwise revealed only through testing a wide variety of experimental conditions. It is now possible to tell at a glance which of these three RNases is involved in the turnover of your favorite mRNA.