In vivo cleavage rules and target repertoire of RNase III in Escherichia coli.

Abstract:

:Bacterial RNase III plays important roles in the processing and degradation of RNA transcripts. A major goal is to identify the cleavage targets of this endoribonuclease at a transcriptome-wide scale and delineate its in vivo cleavage rules. Here we applied to Escherichia coli grown to either exponential or stationary phase a tailored RNA-seq-based technology, which allows transcriptome-wide mapping of RNase III cleavage sites at a nucleotide resolution. Our analysis of the large-scale in vivo cleavage data substantiated the established cleavage pattern of a double cleavage in an intra-molecular stem structure, leaving 2-nt-long 3' overhangs, and refined the base-pairing preferences in the cleavage site vicinity. Intriguingly, we observed that the two stem positions between the cleavage sites are highly base-paired, usually involving at least one G-C or C-G base pair. We present a clear distinction between intra-molecular stem structures that are RNase III substrates and intra-molecular stem structures randomly selected across the transcriptome, emphasizing the in vivo specificity of RNase III. Our study provides a comprehensive map of the cleavage sites in both intra-molecular and inter-molecular duplex substrates, providing novel insights into the involvement of RNase III in post-transcriptional regulation in the bacterial cell.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Altuvia Y,Bar A,Reiss N,Karavani E,Argaman L,Margalit H

doi

10.1093/nar/gky684

subject

Has Abstract

pub_date

2018-11-02 00:00:00

pages

10380-10394

issue

19

eissn

0305-1048

issn

1362-4962

pii

5064785

journal_volume

46

pub_type

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