Two Tandem RNase III Cleavage Sites Determine betT mRNA Stability in Response to Osmotic Stress in Escherichia coli

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While identifying genes regulated by ribonuclease III (RNase III) in Escherichia coli, we observed that steady-state levels of betT mRNA, which encodes a transporter mediating the influx of choline, are dependent on cellular concentrations of RNase III. In the present study, we also observed that steady-state levels of betT mRNA are dependent on RNase III activity upon exposure to osmotic stress, indicating the presence of cis-acting elements controlled by RNase III in betT mRNA. Primer extension analyses of betT mRNA revealed two tandem RNase III cleavage sites in its stem-loop region, which were biochemically confirmed via in vitro cleavage assays. Analyses of cleavage sites suggested the stochastic selection of cleavage sites by RNase III, and mutational analyses indicated that RNase III cleavage at either site individually is insufficient for efficient betT mRNA degradation. In addition, both the half-life and abundance of betT mRNA were significantly increased in association with decreased RNase III activity under hyper-osmotic stress conditions. Our findings demonstrate that betT mRNA stability is controlled by RNase III at the post-transcriptional level under conditions of osmotic stress.
Publisher
PUBLIC LIBRARY SCIENCE
Issue Date
2014-06
Language
English
Article Type
Article
Keywords

GLYCINE BETAINE PATHWAY; SALMONELLA-TYPHIMURIUM; RIBONUCLEASE-III; TRANSPORT-SYSTEM; RIBOSOMAL-RNA; CHOLINE; DEGRADATION; EXPRESSION; GENES; PRODUCTS

Citation

PLOS ONE, v.9, no.6

ISSN
1932-6203
DOI
10.1371/journal.pone.0100520
URI
http://hdl.handle.net/10203/190068
Appears in Collection
CH-Journal Papers(저널논문)
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