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The bacterial enzyme RppH triggers messenger RNA degradation by 5′ pyrophosphate removal

Nature volume 451, pages 355–358 (2008)Cite this article

Abstract

The long-standing assumption that messenger RNA (mRNA) degradation in Escherichia coli begins with endonucleolytic cleavage has been challenged by the recent discovery that RNA decay can be triggered by a prior non-nucleolytic event that marks transcripts for rapid turnover: the rate-determining conversion of the 5′ terminus from a triphosphate to a monophosphate1. This modification creates better substrates for the endonuclease RNase E, whose cleavage activity at internal sites is greatly enhanced when the RNA 5′ end is monophosphorylated2,3. Moreover, it suggests an explanation for the influence of 5′ termini on the endonucleolytic cleavage of primary transcripts, which are triphosphorylated4,5,6,7,8. However, no enzyme capable of removing pyrophosphate from RNA 5′ ends has been identified in any bacterial species. Here we show that the E. coli protein RppH (formerly NudH/YgdP) is the RNA pyrophosphohydrolase that initiates mRNA decay by this 5′-end-dependent pathway. In vitro, RppH efficiently removes pyrophosphate from the 5′ end of triphosphorylated RNA, irrespective of the identity of the 5′-terminal nucleotide. In vivo, it accelerates the degradation of hundreds of E. coli transcripts by converting their triphosphorylated 5′ ends to a more labile monophosphorylated state that can stimulate subsequent ribonuclease cleavage. That the action of the pyrophosphohydrolase is impeded when the 5′ end is structurally sequestered by a stem-loop helps to explain the stabilizing influence of 5′-terminal base pairing on mRNA lifetimes. Together, these findings suggest a possible basis for the effect of RppH and its orthologues on the invasiveness of bacterial pathogens. Interestingly, this master regulator of 5′-end-dependent mRNA degradation in E. coli not only catalyses a process functionally reminiscent of eukaryotic mRNA decapping but also bears an evolutionary relationship to the eukaryotic decapping enzyme Dcp2.

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Figure 1: RNA pyrophosphohydrolase activity of purified RppH.
Figure 2: Triphosphate-to-monophosphate conversion by purified RppH.
Figure 3: RNA pyrophosphohydrolase activity of RppH in E. coli.

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Acknowledgements

We are grateful to D. Guttman for his assistance in the discovery that purified RppH has RNA pyrophosphohydrolase activity. This research was supported by a grant to J.G.B. from the National Institutes of Health.

Author Contributions A.D., H.C. and J.G.B. planned the studies, interpreted the data and wrote the manuscript. A.D. and H.C. performed the experiments.

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Authors and Affiliations

  1. and Department of Microbiology, Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, New York, New York 10016, USA,

    Atilio Deana, Helena Celesnik & Joel G. Belasco

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  1. Atilio Deana
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  3. Joel G. Belasco
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Correspondence to Joel G. Belasco.

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Deana, A., Celesnik, H. & Belasco, J. The bacterial enzyme RppH triggers messenger RNA degradation by 5′ pyrophosphate removal. Nature 451, 355–358 (2008). https://doi.org/10.1038/nature06475

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