An order of magnitude faster AIP1-associated actin disruption than nucleation by the Arp2/3 complex in lamellipodia.

The mechanism of lamellipod actin turnover is still under debate. To clarify the intracellular behavior of the recently-identified actin disruption mechanism, we examined kinetics of AIP1 using fluorescent single-molecule speckle microscopy. AIP1 is thought to cap cofilin-generated actin barbed ends...

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Autores principales:	Takahiro Tsuji, Takushi Miyoshi, Chiharu Higashida, Shuh Narumiya, Naoki Watanabe
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2009
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e2eea70d5a774e5bb6207fb832ad9729
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spelling	oai:doaj.org-article:e2eea70d5a774e5bb6207fb832ad97292021-11-25T06:16:39ZAn order of magnitude faster AIP1-associated actin disruption than nucleation by the Arp2/3 complex in lamellipodia.1932-620310.1371/journal.pone.0004921https://doaj.org/article/e2eea70d5a774e5bb6207fb832ad97292009-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19290054/?tool=EBIhttps://doaj.org/toc/1932-6203The mechanism of lamellipod actin turnover is still under debate. To clarify the intracellular behavior of the recently-identified actin disruption mechanism, we examined kinetics of AIP1 using fluorescent single-molecule speckle microscopy. AIP1 is thought to cap cofilin-generated actin barbed ends. Here we demonstrate a reduction in actin-associated AIP1 in lamellipodia of cells overexpressing LIM-kinase. Moreover, actin-associated AIP1 was rapidly abolished by jasplakinolide, which concurrently blocked the F-actin-cofilin interaction. Jasplakinolide also slowed dissociation of AIP1, which is analogous to the effect of this drug on capping protein. These findings provide in vivo evidence of the association of AIP1 with barbed ends generated by cofilin-catalyzed filament disruption. Single-molecule observation found distribution of F-actin-associated AIP1 throughout lamellipodia, and revealed even faster dissociation of AIP1 than capping protein. The estimated overall AIP1-associated actin disruption rate, 1.8 microM/s, was one order of magnitude faster than Arp2/3 complex-catalyzed actin nucleation in lamellipodia. This rate does not suffice the filament severing rate predicted in our previous high frequency filament severing-annealing hypothesis. Our data together with recent biochemical studies imply barbed end-preferred frequent filament disruption. Frequent generation of AIP1-associated barbed ends and subsequent release of AIP1 may be the mechanism that facilitates previously observed ubiquitous actin polymerization throughout lamellipodia.Takahiro TsujiTakushi MiyoshiChiharu HigashidaShuh NarumiyaNaoki WatanabePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 4, Iss 3, p e4921 (2009)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Takahiro Tsuji Takushi Miyoshi Chiharu Higashida Shuh Narumiya Naoki Watanabe An order of magnitude faster AIP1-associated actin disruption than nucleation by the Arp2/3 complex in lamellipodia.
description	The mechanism of lamellipod actin turnover is still under debate. To clarify the intracellular behavior of the recently-identified actin disruption mechanism, we examined kinetics of AIP1 using fluorescent single-molecule speckle microscopy. AIP1 is thought to cap cofilin-generated actin barbed ends. Here we demonstrate a reduction in actin-associated AIP1 in lamellipodia of cells overexpressing LIM-kinase. Moreover, actin-associated AIP1 was rapidly abolished by jasplakinolide, which concurrently blocked the F-actin-cofilin interaction. Jasplakinolide also slowed dissociation of AIP1, which is analogous to the effect of this drug on capping protein. These findings provide in vivo evidence of the association of AIP1 with barbed ends generated by cofilin-catalyzed filament disruption. Single-molecule observation found distribution of F-actin-associated AIP1 throughout lamellipodia, and revealed even faster dissociation of AIP1 than capping protein. The estimated overall AIP1-associated actin disruption rate, 1.8 microM/s, was one order of magnitude faster than Arp2/3 complex-catalyzed actin nucleation in lamellipodia. This rate does not suffice the filament severing rate predicted in our previous high frequency filament severing-annealing hypothesis. Our data together with recent biochemical studies imply barbed end-preferred frequent filament disruption. Frequent generation of AIP1-associated barbed ends and subsequent release of AIP1 may be the mechanism that facilitates previously observed ubiquitous actin polymerization throughout lamellipodia.
format	article
author	Takahiro Tsuji Takushi Miyoshi Chiharu Higashida Shuh Narumiya Naoki Watanabe
author_facet	Takahiro Tsuji Takushi Miyoshi Chiharu Higashida Shuh Narumiya Naoki Watanabe
author_sort	Takahiro Tsuji
title	An order of magnitude faster AIP1-associated actin disruption than nucleation by the Arp2/3 complex in lamellipodia.
title_short	An order of magnitude faster AIP1-associated actin disruption than nucleation by the Arp2/3 complex in lamellipodia.
title_full	An order of magnitude faster AIP1-associated actin disruption than nucleation by the Arp2/3 complex in lamellipodia.
title_fullStr	An order of magnitude faster AIP1-associated actin disruption than nucleation by the Arp2/3 complex in lamellipodia.
title_full_unstemmed	An order of magnitude faster AIP1-associated actin disruption than nucleation by the Arp2/3 complex in lamellipodia.
title_sort	order of magnitude faster aip1-associated actin disruption than nucleation by the arp2/3 complex in lamellipodia.
publisher	Public Library of Science (PLoS)
publishDate	2009
url	https://doaj.org/article/e2eea70d5a774e5bb6207fb832ad9729
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An order of magnitude faster AIP1-associated actin disruption than nucleation by the Arp2/3 complex in lamellipodia.

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