原创力文档an upstream hfq binding site in the fhla mrna leader region facilitates the oxys-fhla interaction上游hfq的结合位点fhla mrna领袖地区促进oxys-fhla交互.pdfVIP
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an upstream hfq binding site in the fhla mrna leader region facilitates the oxys-fhla interaction上游hfq的结合位点fhla mrna领袖地区促进oxys-fhla交互
An Upstream Hfq Binding Site in the fhlA mRNA Leader
Region Facilitates the OxyS-fhlA Interaction
Nilshad N. Salim, Andrew L. Feig*
Department of Chemistry, Wayne State University, Detroit, Michigan, United States of America
Abstract
Background: To survive, bacteria must be able to adapt to environmental stresses. Small regulatory RNAs have been
implicated as intermediates in a variety of stress-response pathways allowing dynamic gene regulation. The RNA binding
protein Hfq facilitates this process in many cases, helping sRNAs base pair with their target mRNAs and initiate gene
regulation. Although Hfq has been identified as a critical component in many RNPs, the manner by which Hfq controls these
interactions is not known.
Methodology/Principal Findings: To test the requirement of Hfq in these mRNA-sRNA complexes, the OxyS-fhlA system
was used as a model. OxyS is induced in response to oxidative stress and down regulates the translation of fhlA, a gene
encoding a transcriptional activator for formate metabolism. Biophysical characterization of this system previously used a
minimal construct of the fhlA mRNA which inadvertently removed a critical element within the leader sequence of this
mRNA that effected thermodynamics and kinetics for the interaction with Hfq.
Conclusions/Significance: Herein, we report thermodynamic, kinetic and structural mapping studies during binary and
ternary complex formation between Hfq, OxyS and fhlA mRNA. Hfq binds fhlA mRNA using both the proximal and distal
surfaces and stimulates association kinetics between the sRNA and mRNA but remains bound to fhlA forming a ternary
complex. The upstream Hfq binding element within fhlA is similar to (ARN)x elements recently identified in other mRNAs
regulated by Hfq. This work leads to a kinetic model fo
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