In vitro analysis of the interaction between the small RNA SR1 and its primary target ahrC mRNA.

Abstract:

:Small regulatory RNAs (sRNAs) from bacterial chromosomes became the focus of research over the past five years. However, relatively little is known in terms of structural requirements, kinetics of interaction with their targets and degradation in contrast to well-studied plasmid-encoded antisense RNAs. Here, we present a detailed in vitro analysis of SR1, a sRNA of Bacillus subtilis that is involved in regulation of arginine catabolism by basepairing with its target, ahrC mRNA. The secondary structures of SR1 species of different lengths and of the SR1/ahrC RNA complex were determined and functional segments required for complex formation narrowed down. The initial contact between SR1 and its target was shown to involve the 5' part of the SR1 terminator stem and a region 100 bp downstream from the ahrC transcriptional start site. Toeprinting studies and secondary structure probing of the ahrC/SR1 complex indicated that SR1 inhibits translation initiation by inducing structural changes downstream from the ahrC RBS. Furthermore, it was demonstrated that Hfq, which binds both SR1 and ahrC RNA was not required to promote ahrC/SR1 complex formation but to enable the translation of ahrC mRNA. The intracellular concentrations of SR1 were calculated under different growth conditions.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Heidrich N,Moll I,Brantl S

doi

10.1093/nar/gkm439

subject

Has Abstract

pub_date

2007-01-01 00:00:00

pages

4331-46

issue

13

eissn

0305-1048

issn

1362-4962

pii

gkm439

journal_volume

35

pub_type

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