Ribonuclease E is a 5′-end-dependent endonuclease (original) (raw)

Nature volume 395, pages 720–724 (1998)Cite this article

Abstract

The selective degradation of messenger RNAs enables cells to regulate the levels of particular mRNAs in response to changes in the environment. Ribonuclease (RNase) E (ref. 1), a single-strand-specific endonuclease2,3,4 that is found in a multi-enzyme complex known as the ‘degradosome’5,6,7, initiates the degradation of many mRNAs in Escherichia coli3,8,9. Its relative lack of sequence specificity and the presence of many potential cleavage sites in mRNA substrates2,3 cannot explain why mRNA decay frequently proceeds in a net 5′-to-3′ direction9,10,11. I have prepared covalently closed circular derivatives of natural substrates, the rpsT mRNA encoding ribosomal protein S20 (ref. 2) and the 9S precursor to 5S ribosomal RNA1,12, and find that these derivatives are considerably more resistant to cleavage in vitro by RNase E than are linear molecules. Moreover, antisense oligo-deoxynucleotides complementary to the 5′ end of linear substrates significantly reduce the latter's susceptibility to attack by RNase E. Finally, natural substrates with terminal 5′-triphosphate groups are poorly cleaved by RNase E in vitro, whereas 5′ monophosphorylated substrates are strongly preferred (compare with ref. 13). These results show that RNase E has inherent vectorial properties, with its activity depending on the 5′ end of its substrates; this can account for the direction of mRNA decay in E. coli, the phenomenon of ‘all or none’ mRNA decay, and the stabilization provided by 5′ stem–loop structures14,15,16,17.

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Acknowledgements

I thank G. Coburn for preparing degradosomes, A. Prud'homme-Genereux for helping to prepare Fig. 4, members of my laboratory for comments, and the MRC of Canada for an operating grant.

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  1. Department of Biochemsitry & Molecular Biology, University of British Columbia, 2146 Health Sciences Mall, Vancouver, V6T 1Z3, British Columbia, Canada
    George A. Mackie

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Mackie, G. Ribonuclease E is a 5′-end-dependent endonuclease.Nature 395, 720–724 (1998). https://doi.org/10.1038/27246

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