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Single-molecule visualization of a formin-capping protein ‘decision complex' at the actin filament barbed end

Precise control of actin filament length is essential to many cellular processes. Formins processively elongate filaments, whereas capping protein (CP) binds to barbed ends and arrests polymerization. While genetic and biochemical evidence has indicated that these two proteins function antagonistica...

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Autores principales: Bombardier, Jeffrey P., Eskin, Julian A., Jaiswal, Richa, Corrêa, Ivan R., Xu, Ming-Qun, Goode, Bruce L., Gelles, Jeff
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Pub. Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4660045/
https://www.ncbi.nlm.nih.gov/pubmed/26566078
http://dx.doi.org/10.1038/ncomms9707
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author Bombardier, Jeffrey P.
Eskin, Julian A.
Jaiswal, Richa
Corrêa, Ivan R.
Xu, Ming-Qun
Goode, Bruce L.
Gelles, Jeff
author_facet Bombardier, Jeffrey P.
Eskin, Julian A.
Jaiswal, Richa
Corrêa, Ivan R.
Xu, Ming-Qun
Goode, Bruce L.
Gelles, Jeff
author_sort Bombardier, Jeffrey P.
collection PubMed
description Precise control of actin filament length is essential to many cellular processes. Formins processively elongate filaments, whereas capping protein (CP) binds to barbed ends and arrests polymerization. While genetic and biochemical evidence has indicated that these two proteins function antagonistically, the mechanism underlying the antagonism has remained unresolved. Here we use multi-wavelength single-molecule fluorescence microscopy to observe the fully reversible formation of a long-lived ‘decision complex' in which a CP dimer and a dimer of the formin mDia1 simultaneously bind the barbed end. Further, mDia1 displaced from the barbed end by CP can randomly slide along the filament and later return to the barbed end to re-form the complex. Quantitative kinetic analysis reveals that the CP-mDia1 antagonism that we observe in vitro occurs through the decision complex. Our observations suggest new molecular mechanisms for the control of actin filament length and for the capture of filament barbed ends in cells.
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spelling pubmed-46600452015-12-04 Single-molecule visualization of a formin-capping protein ‘decision complex' at the actin filament barbed end Bombardier, Jeffrey P. Eskin, Julian A. Jaiswal, Richa Corrêa, Ivan R. Xu, Ming-Qun Goode, Bruce L. Gelles, Jeff Nat Commun Article Precise control of actin filament length is essential to many cellular processes. Formins processively elongate filaments, whereas capping protein (CP) binds to barbed ends and arrests polymerization. While genetic and biochemical evidence has indicated that these two proteins function antagonistically, the mechanism underlying the antagonism has remained unresolved. Here we use multi-wavelength single-molecule fluorescence microscopy to observe the fully reversible formation of a long-lived ‘decision complex' in which a CP dimer and a dimer of the formin mDia1 simultaneously bind the barbed end. Further, mDia1 displaced from the barbed end by CP can randomly slide along the filament and later return to the barbed end to re-form the complex. Quantitative kinetic analysis reveals that the CP-mDia1 antagonism that we observe in vitro occurs through the decision complex. Our observations suggest new molecular mechanisms for the control of actin filament length and for the capture of filament barbed ends in cells. Nature Pub. Group 2015-11-13 /pmc/articles/PMC4660045/ /pubmed/26566078 http://dx.doi.org/10.1038/ncomms9707 Text en Copyright © 2015, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Bombardier, Jeffrey P.
Eskin, Julian A.
Jaiswal, Richa
Corrêa, Ivan R.
Xu, Ming-Qun
Goode, Bruce L.
Gelles, Jeff
Single-molecule visualization of a formin-capping protein ‘decision complex' at the actin filament barbed end
title Single-molecule visualization of a formin-capping protein ‘decision complex' at the actin filament barbed end
title_full Single-molecule visualization of a formin-capping protein ‘decision complex' at the actin filament barbed end
title_fullStr Single-molecule visualization of a formin-capping protein ‘decision complex' at the actin filament barbed end
title_full_unstemmed Single-molecule visualization of a formin-capping protein ‘decision complex' at the actin filament barbed end
title_short Single-molecule visualization of a formin-capping protein ‘decision complex' at the actin filament barbed end
title_sort single-molecule visualization of a formin-capping protein ‘decision complex' at the actin filament barbed end
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4660045/
https://www.ncbi.nlm.nih.gov/pubmed/26566078
http://dx.doi.org/10.1038/ncomms9707
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