Abstract
The sRNA RprA is known to activate rpoS translation in E. coli in an osmolarity-dependent manner. We asked whether RprA stability contributes to osmolarity-dependent regulation and how the RNA binding protein Hfq and the major E. coli endonucleases contribute to this turn-over. The study reveals that osmolarity-dependent turn-over of RprA indeed contributes to its osmolarity-dependent abundance. RprA is stabilized by the RNA chaperone Hfq and in absence of Hfq its turn-over is no longer osmolarity-dependent. The stability of the RprA target mRNA rpoS shows a lower extent of osmolarity dependence, which differs from the profile observed for RprA. Thus, the effect of sucrose is specific for individual RNAs. We can attribute a role of the endoribonuclease RNase E in turn-over of RprA and an indirect effect of the endoribonuclease III in vivo. In addition, RprA is stabilized by the presence of rpoS suggesting that hybrid formation with its target may protect it against ribonucleases. In vitro RprA is cleaved by the RNase E containing degradosome and by RNase III and rpoS interferes with RNase III cleavage. We also show that temperature affects the stabilities of the sRNAs binding to rpoS and of rpoS mRNA itself differentially and that higher stability of DsrA with decreasing temperature may contribute to its high abundance at lower temperatures. This study demonstrates that environmental parameters can affect the stability of sRNAs and consequently their abundance.
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Acknowledgments
We thank Christian Lassek and Joachim Berk for assistance in performing the experiments. We also thank Prof. Poul Valentin-Hansen, University of Southern Denmark, for kindly providing us antibodies against Hfq. This work was supported by Deutsche Forschungsgemeinschaft (DFG Kl563/19-1) and a DAAD fellowship to RM.
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Communicated by D. Andersson.
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Madhugiri, R., Basineni, S.R. & Klug, G. Turn-over of the small non-coding RNA RprA in E. coli is influenced by osmolarity. Mol Genet Genomics 284, 307–318 (2010). https://doi.org/10.1007/s00438-010-0568-x
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DOI: https://doi.org/10.1007/s00438-010-0568-x