Stretching actin filaments within cells enhances their affinity for the myosin II motor domain.
To test the hypothesis that the myosin II motor domain (S1) preferentially binds to specific subsets of actin filaments in vivo, we expressed GFP-fused S1 with mutations that enhanced its affinity for actin in Dictyostelium cells. Consistent with the hypothesis, the GFP-S1 mutants were localized alo...
Main Authors: | , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Public Library of Science (PLoS)
2011-01-01
|
Series: | PLoS ONE |
Online Access: | http://europepmc.org/articles/PMC3192770?pdf=render |
id |
doaj-7b5b8dbb8c1043bbad6527b7501f1281 |
---|---|
record_format |
Article |
spelling |
doaj-7b5b8dbb8c1043bbad6527b7501f12812020-11-25T01:44:56ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-01610e2620010.1371/journal.pone.0026200Stretching actin filaments within cells enhances their affinity for the myosin II motor domain.Taro Q P UyedaYoshiaki IwadateNobuhisa UmekiAkira NagasakiShigehiko YumuraTo test the hypothesis that the myosin II motor domain (S1) preferentially binds to specific subsets of actin filaments in vivo, we expressed GFP-fused S1 with mutations that enhanced its affinity for actin in Dictyostelium cells. Consistent with the hypothesis, the GFP-S1 mutants were localized along specific portions of the cell cortex. Comparison with rhodamine-phalloidin staining in fixed cells demonstrated that the GFP-S1 probes preferentially bound to actin filaments in the rear cortex and cleavage furrows, where actin filaments are stretched by interaction with endogenous myosin II filaments. The GFP-S1 probes were similarly enriched in the cortex stretched passively by traction forces in the absence of myosin II or by external forces using a microcapillary. The preferential binding of GFP-S1 mutants to stretched actin filaments did not depend on cortexillin I or PTEN, two proteins previously implicated in the recruitment of myosin II filaments to stretched cortex. These results suggested that it is the stretching of the actin filaments itself that increases their affinity for the myosin II motor domain. In contrast, the GFP-fused myosin I motor domain did not localize to stretched actin filaments, which suggests different preferences of the motor domains for different structures of actin filaments play a role in distinct intracellular localizations of myosin I and II. We propose a scheme in which the stretching of actin filaments, the preferential binding of myosin II filaments to stretched actin filaments, and myosin II-dependent contraction form a positive feedback loop that contributes to the stabilization of cell polarity and to the responsiveness of the cells to external mechanical stimuli.http://europepmc.org/articles/PMC3192770?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Taro Q P Uyeda Yoshiaki Iwadate Nobuhisa Umeki Akira Nagasaki Shigehiko Yumura |
spellingShingle |
Taro Q P Uyeda Yoshiaki Iwadate Nobuhisa Umeki Akira Nagasaki Shigehiko Yumura Stretching actin filaments within cells enhances their affinity for the myosin II motor domain. PLoS ONE |
author_facet |
Taro Q P Uyeda Yoshiaki Iwadate Nobuhisa Umeki Akira Nagasaki Shigehiko Yumura |
author_sort |
Taro Q P Uyeda |
title |
Stretching actin filaments within cells enhances their affinity for the myosin II motor domain. |
title_short |
Stretching actin filaments within cells enhances their affinity for the myosin II motor domain. |
title_full |
Stretching actin filaments within cells enhances their affinity for the myosin II motor domain. |
title_fullStr |
Stretching actin filaments within cells enhances their affinity for the myosin II motor domain. |
title_full_unstemmed |
Stretching actin filaments within cells enhances their affinity for the myosin II motor domain. |
title_sort |
stretching actin filaments within cells enhances their affinity for the myosin ii motor domain. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2011-01-01 |
description |
To test the hypothesis that the myosin II motor domain (S1) preferentially binds to specific subsets of actin filaments in vivo, we expressed GFP-fused S1 with mutations that enhanced its affinity for actin in Dictyostelium cells. Consistent with the hypothesis, the GFP-S1 mutants were localized along specific portions of the cell cortex. Comparison with rhodamine-phalloidin staining in fixed cells demonstrated that the GFP-S1 probes preferentially bound to actin filaments in the rear cortex and cleavage furrows, where actin filaments are stretched by interaction with endogenous myosin II filaments. The GFP-S1 probes were similarly enriched in the cortex stretched passively by traction forces in the absence of myosin II or by external forces using a microcapillary. The preferential binding of GFP-S1 mutants to stretched actin filaments did not depend on cortexillin I or PTEN, two proteins previously implicated in the recruitment of myosin II filaments to stretched cortex. These results suggested that it is the stretching of the actin filaments itself that increases their affinity for the myosin II motor domain. In contrast, the GFP-fused myosin I motor domain did not localize to stretched actin filaments, which suggests different preferences of the motor domains for different structures of actin filaments play a role in distinct intracellular localizations of myosin I and II. We propose a scheme in which the stretching of actin filaments, the preferential binding of myosin II filaments to stretched actin filaments, and myosin II-dependent contraction form a positive feedback loop that contributes to the stabilization of cell polarity and to the responsiveness of the cells to external mechanical stimuli. |
url |
http://europepmc.org/articles/PMC3192770?pdf=render |
work_keys_str_mv |
AT taroqpuyeda stretchingactinfilamentswithincellsenhancestheiraffinityforthemyosiniimotordomain AT yoshiakiiwadate stretchingactinfilamentswithincellsenhancestheiraffinityforthemyosiniimotordomain AT nobuhisaumeki stretchingactinfilamentswithincellsenhancestheiraffinityforthemyosiniimotordomain AT akiranagasaki stretchingactinfilamentswithincellsenhancestheiraffinityforthemyosiniimotordomain AT shigehikoyumura stretchingactinfilamentswithincellsenhancestheiraffinityforthemyosiniimotordomain |
_version_ |
1725026133753200640 |