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A positive charge region of Salmonella FliI is required for ATPase formation and efficient flagellar protein export

The FliH(2)FliI complex is thought to pilot flagellar subunit proteins from the cytoplasm to the transmembrane export gate complex for flagellar assembly in Salmonella enterica. FliI also forms a homo-hexamer to hydrolyze ATP, thereby activating the export gate complex to become an active protein tr...

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Autores principales: Kinoshita, Miki, Namba, Keiichi, Minamino, Tohru
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8041783/
https://www.ncbi.nlm.nih.gov/pubmed/33846530
http://dx.doi.org/10.1038/s42003-021-01980-y
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author Kinoshita, Miki
Namba, Keiichi
Minamino, Tohru
author_facet Kinoshita, Miki
Namba, Keiichi
Minamino, Tohru
author_sort Kinoshita, Miki
collection PubMed
description The FliH(2)FliI complex is thought to pilot flagellar subunit proteins from the cytoplasm to the transmembrane export gate complex for flagellar assembly in Salmonella enterica. FliI also forms a homo-hexamer to hydrolyze ATP, thereby activating the export gate complex to become an active protein transporter. However, it remains unknown how this activation occurs. Here we report the role of a positively charged cluster formed by Arg-26, Arg-27, Arg-33, Arg-76 and Arg-93 of FliI in flagellar protein export. We show that Arg-33 and Arg-76 are involved in FliI ring formation and that the fliI(R26A/R27A/R33A/R76A/R93A) mutant requires the presence of FliH to fully exert its export function. We observed that gain-of-function mutations in FlhB increased the probability of substrate entry into the export gate complex, thereby restoring the export function of the ∆fliH fliI(R26A/R27A/R33A/R76A/R93A) mutant. We suggest that the positive charge cluster of FliI is responsible not only for well-regulated hexamer assembly but also for substrate entry into the gate complex.
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spelling pubmed-80417832021-04-28 A positive charge region of Salmonella FliI is required for ATPase formation and efficient flagellar protein export Kinoshita, Miki Namba, Keiichi Minamino, Tohru Commun Biol Article The FliH(2)FliI complex is thought to pilot flagellar subunit proteins from the cytoplasm to the transmembrane export gate complex for flagellar assembly in Salmonella enterica. FliI also forms a homo-hexamer to hydrolyze ATP, thereby activating the export gate complex to become an active protein transporter. However, it remains unknown how this activation occurs. Here we report the role of a positively charged cluster formed by Arg-26, Arg-27, Arg-33, Arg-76 and Arg-93 of FliI in flagellar protein export. We show that Arg-33 and Arg-76 are involved in FliI ring formation and that the fliI(R26A/R27A/R33A/R76A/R93A) mutant requires the presence of FliH to fully exert its export function. We observed that gain-of-function mutations in FlhB increased the probability of substrate entry into the export gate complex, thereby restoring the export function of the ∆fliH fliI(R26A/R27A/R33A/R76A/R93A) mutant. We suggest that the positive charge cluster of FliI is responsible not only for well-regulated hexamer assembly but also for substrate entry into the gate complex. Nature Publishing Group UK 2021-04-12 /pmc/articles/PMC8041783/ /pubmed/33846530 http://dx.doi.org/10.1038/s42003-021-01980-y Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Kinoshita, Miki
Namba, Keiichi
Minamino, Tohru
A positive charge region of Salmonella FliI is required for ATPase formation and efficient flagellar protein export
title A positive charge region of Salmonella FliI is required for ATPase formation and efficient flagellar protein export
title_full A positive charge region of Salmonella FliI is required for ATPase formation and efficient flagellar protein export
title_fullStr A positive charge region of Salmonella FliI is required for ATPase formation and efficient flagellar protein export
title_full_unstemmed A positive charge region of Salmonella FliI is required for ATPase formation and efficient flagellar protein export
title_short A positive charge region of Salmonella FliI is required for ATPase formation and efficient flagellar protein export
title_sort positive charge region of salmonella flii is required for atpase formation and efficient flagellar protein export
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8041783/
https://www.ncbi.nlm.nih.gov/pubmed/33846530
http://dx.doi.org/10.1038/s42003-021-01980-y
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