In Vivo Cleavage Map Illuminates the Central Role of RNase E in Coding and Non-coding RNA Pathways
Förstner, Konrad U
Luisi, Bonaventura Francesco
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Chao, Y., Li, L., Girodat, D., Förstner, K. U., Said, N., Corcoran, C., Śmiga, M., et al. (2017). In Vivo Cleavage Map Illuminates the Central Role of RNase E in Coding and Non-coding RNA Pathways. Molecular Cell, 65 (1), 39-51. https://doi.org/10.1016/j.molcel.2016.11.002
Understanding RNA processing and turnover requires knowledge of cleavages by major endoribonucleases within a living cell. We have employed TIER-seq (transiently inactivating an endoribonuclease followed by RNA-seq) to profile cleavage products of the essential endoribonuclease RNase E in Salmonella enterica. A dominating cleavage signature is the location of a uridine two nucleotides downstream in a single-stranded segment, which we rationalize structurally as a key recognition determinant that may favor RNase E catalysis. Our results suggest a prominent biogenesis pathway for bacterial regulatory small RNAs whereby RNase E acts together with the RNA chaperone Hfq to liberate stable 3′ fragments from various precursor RNAs. Recapitulating this process in vitro, Hfq guides RNase E cleavage of a representative small-RNA precursor for interaction with a mRNA target. In vivo, the processing is required for target regulation. Our findings reveal a general maturation mechanism for a major class of post-transcriptional regulators.
This study was funded by DFG ( Vo875/14-1 ) and BioSysNet grants. B.F.L. is supported by the Wellcome Trust. K.P. was supported by the Human Frontiers Science Program (CDA00024/2016-C) .
External DOI: https://doi.org/10.1016/j.molcel.2016.11.002
This record's URL: https://www.repository.cam.ac.uk/handle/1810/270005
Attribution 4.0 International
Licence URL: http://creativecommons.org/licenses/by/4.0/
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