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Published on: August 20, 2014
Metastable Folding of Bacillus subtilis glmS Ribozyme Modulates Turnover by RNase J1
1T.C. Jenkins Department of Biophysics, Johns Hopkins University, 3400 N. Charles St., Baltimore, MD 21218, USA.
Abstract:
Rapid turnover of glmS mRNA in Bacillus subtilis by 5'-3' exoribonuclease RNase J is essential for feedback regulation of glucosamine-6-phosphate (GlcN6P) synthase expression, upon self-cleavage of a GlcN6P-activated ribozyme in the glmS 5' UTR. We used biochemical assays and single molecule fluorescence microscopy to show that initiation of RNase J decay is inefficient and requires approximately 15 5' unpaired nucleotides to form a processive exonuclease complex that is insensitive to downstream RNA structure. When stably folded, the cleaved glmS ribozyme blocks RNase J initiation. However, co-transcriptional ribozyme cleavage and physiological Mg2+ levels increase decay by weakening the ribozyme structure. At 22 °C, the processive velocity of RNase J, 23 ± 8 nt/s, is equal to or faster than transcription, indicating that RNase J has the potential to catch elongating polymerases. The results show how the folding stability of 5' mRNA structure contributes to RNase J recognition and the control of mRNA half-life.
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