title: Rapid cleavage of RNA by RNase E in the absence of 5' monophosphate stimulation.
abstract: The best characterized pathway for the initiation of mRNA degradation in Escherichia coli involves the removal of the 5'-terminal pyrophosphate to generate a monophosphate group that stimulates endonucleolytic cleavage by RNase E. We show here however, using well-characterized oligonucleotide substrates and mRNA transcripts, that RNase E can cleave certain RNAs rapidly without requiring a 5'-monophosphorylated end. Moreover, the minimum substrate requirement for this mode of cleavage, which can be categorized as 'direct' or 'internal' entry, appears to be multiple single-stranded segments in a conformational context that allows their simultaneous interaction with RNase E. While previous work has alluded to the existence of a 5' end-independent mechanism of mRNA degradation, the relative simplicity of the requirements identified here for direct entry suggests that it could represent a major means by which mRNA degradation is initiated in E. coli and other organisms that contain homologues of RNase E. Our results have implications for the interplay of translation and mRNA degradation and models of gene regulation by small non-coding RNAs.
authors.affiliation: Astbury Centre for Structural Molecular Biology, Faculty of Biological Sciences, University of Leeds, LS2 9JT, England, UK.
authors.forename: Louise
authors.initials: L
authors.lastname: Kime
authors.forename: Stefanie S
authors.initials: SS
authors.lastname: Jourdan
authors.forename: Jonathan A
authors.initials: JA
authors.lastname: Stead
authors.forename: Ana
authors.initials: A
authors.lastname: Hidalgo-Sastre
authors.forename: Kenneth J
authors.initials: KJ
authors.lastname: McDowall
mesh_terms: D001483:Base Sequence; D015226:Dinucleoside Phosphates / Q000378:metabolism*; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D020871:RNA Stability; D012329:RNA, Bacterial / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D015226:Dinucleoside Phosphates; D029968:Escherichia coli Proteins; D012329:RNA, Bacterial; D004722:Endoribonucleases; C022537:ribonuclease E