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Binding of Campylobacter jejuni FliW Adjacent to the CsrA RNA-Binding Pockets Modulates CsrA Regulatory Activity

Campylobacter jejuni CsrA is an mRNA-binding, post-transcriptional regulator that controls many metabolic- and virulence-related characteristics of this important pathogen. In contrast to E. coli CsrA, whose activity is modulated by binding to small non-coding RNAs (sRNAs), C. jejuni CsrA activity i...

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Autores principales: Bogacz, Marek, El Abbar, Faiha M., Cox, Claudia A., Li, Jiaqi, Fiedler, Jarred S., Tran, Lynn K. H., Tran, Paul M. H., Daugherty, C. Luke, Blake, Kate H., Wang, Zhirui, Azadi, Parastoo, Thompson, Stuart A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7829508/
https://www.ncbi.nlm.nih.gov/pubmed/33505360
http://dx.doi.org/10.3389/fmicb.2020.531596
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author Bogacz, Marek
El Abbar, Faiha M.
Cox, Claudia A.
Li, Jiaqi
Fiedler, Jarred S.
Tran, Lynn K. H.
Tran, Paul M. H.
Daugherty, C. Luke
Blake, Kate H.
Wang, Zhirui
Azadi, Parastoo
Thompson, Stuart A.
author_facet Bogacz, Marek
El Abbar, Faiha M.
Cox, Claudia A.
Li, Jiaqi
Fiedler, Jarred S.
Tran, Lynn K. H.
Tran, Paul M. H.
Daugherty, C. Luke
Blake, Kate H.
Wang, Zhirui
Azadi, Parastoo
Thompson, Stuart A.
author_sort Bogacz, Marek
collection PubMed
description Campylobacter jejuni CsrA is an mRNA-binding, post-transcriptional regulator that controls many metabolic- and virulence-related characteristics of this important pathogen. In contrast to E. coli CsrA, whose activity is modulated by binding to small non-coding RNAs (sRNAs), C. jejuni CsrA activity is controlled by binding to the CsrA antagonist FliW. In this study, we identified the FliW binding site on CsrA. Deletion of the C-terminus of C. jejuni CsrA, which is extended relative to sRNA-binding CsrA proteins, abrogated FliW binding. Bacterial two-hybrid experiments were used to assess the interaction of FliW with wild-type CsrA and mutants thereof, in which every amino acid was individually mutated. Two CsrA mutations (V51A and N55A) resulted in a significant decrease in FliW binding. The V51A and N55A mutants also showed a decrease in CsrA-FliW complex formation, as assessed by size-exclusion chromatography and surface plasmon resonance. These residues were highly conserved in bacterial species containing CsrA orthologs whose activities are predicted to be regulated by FliW. The location of FliW binding was immediately adjacent to the two RNA-binding sites of the CsrA homodimer, suggesting the model that FliW binding to CsrA modulates its ability to bind to its mRNA targets either by steric hindrance, electrostatic repulsion, or by altering the overall structure of the RNA-binding sites.
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spelling pubmed-78295082021-01-26 Binding of Campylobacter jejuni FliW Adjacent to the CsrA RNA-Binding Pockets Modulates CsrA Regulatory Activity Bogacz, Marek El Abbar, Faiha M. Cox, Claudia A. Li, Jiaqi Fiedler, Jarred S. Tran, Lynn K. H. Tran, Paul M. H. Daugherty, C. Luke Blake, Kate H. Wang, Zhirui Azadi, Parastoo Thompson, Stuart A. Front Microbiol Microbiology Campylobacter jejuni CsrA is an mRNA-binding, post-transcriptional regulator that controls many metabolic- and virulence-related characteristics of this important pathogen. In contrast to E. coli CsrA, whose activity is modulated by binding to small non-coding RNAs (sRNAs), C. jejuni CsrA activity is controlled by binding to the CsrA antagonist FliW. In this study, we identified the FliW binding site on CsrA. Deletion of the C-terminus of C. jejuni CsrA, which is extended relative to sRNA-binding CsrA proteins, abrogated FliW binding. Bacterial two-hybrid experiments were used to assess the interaction of FliW with wild-type CsrA and mutants thereof, in which every amino acid was individually mutated. Two CsrA mutations (V51A and N55A) resulted in a significant decrease in FliW binding. The V51A and N55A mutants also showed a decrease in CsrA-FliW complex formation, as assessed by size-exclusion chromatography and surface plasmon resonance. These residues were highly conserved in bacterial species containing CsrA orthologs whose activities are predicted to be regulated by FliW. The location of FliW binding was immediately adjacent to the two RNA-binding sites of the CsrA homodimer, suggesting the model that FliW binding to CsrA modulates its ability to bind to its mRNA targets either by steric hindrance, electrostatic repulsion, or by altering the overall structure of the RNA-binding sites. Frontiers Media S.A. 2021-01-11 /pmc/articles/PMC7829508/ /pubmed/33505360 http://dx.doi.org/10.3389/fmicb.2020.531596 Text en Copyright © 2021 Bogacz, El Abbar, Cox, Li, Fiedler, Tran, Tran, Daugherty, Blake, Wang, Azadi and Thompson. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Bogacz, Marek
El Abbar, Faiha M.
Cox, Claudia A.
Li, Jiaqi
Fiedler, Jarred S.
Tran, Lynn K. H.
Tran, Paul M. H.
Daugherty, C. Luke
Blake, Kate H.
Wang, Zhirui
Azadi, Parastoo
Thompson, Stuart A.
Binding of Campylobacter jejuni FliW Adjacent to the CsrA RNA-Binding Pockets Modulates CsrA Regulatory Activity
title Binding of Campylobacter jejuni FliW Adjacent to the CsrA RNA-Binding Pockets Modulates CsrA Regulatory Activity
title_full Binding of Campylobacter jejuni FliW Adjacent to the CsrA RNA-Binding Pockets Modulates CsrA Regulatory Activity
title_fullStr Binding of Campylobacter jejuni FliW Adjacent to the CsrA RNA-Binding Pockets Modulates CsrA Regulatory Activity
title_full_unstemmed Binding of Campylobacter jejuni FliW Adjacent to the CsrA RNA-Binding Pockets Modulates CsrA Regulatory Activity
title_short Binding of Campylobacter jejuni FliW Adjacent to the CsrA RNA-Binding Pockets Modulates CsrA Regulatory Activity
title_sort binding of campylobacter jejuni fliw adjacent to the csra rna-binding pockets modulates csra regulatory activity
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7829508/
https://www.ncbi.nlm.nih.gov/pubmed/33505360
http://dx.doi.org/10.3389/fmicb.2020.531596
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