Traction forces of fibroblasts are regulated by the Rho-dependent kinase but not by the myosin light chain kinase

被引:99
作者
Beningo, Karen A.
Hamao, Kozue
Dembo, Micah
Wang, Yu-li [1 ]
Hosoya, Hiroshi
机构
[1] Univ Massachusetts, Sch Med, Dept Physiol, Worcester, MA 01605 USA
[2] Wayne State Univ, Dept Biol Sci, Detroit, MI 48202 USA
[3] Boston Univ, Dept Biomed Engn, Boston, MA 02215 USA
[4] Hiroshima Univ, Dept Biol Sci, Higashihiroshima 7398256, Japan
关键词
cell migration; myosin; myosin light chain kinase; rho; small GTPases; focal adhesions; mechanotransduction;
D O I
10.1016/j.abb.2006.09.025
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Adhesive cells show complex mechanical interactions with the substrate, however the exact mechanism of such interactions, termed traction forces, is still unclear. To address this question we have measured traction forces of fibroblasts treated with agents that affect the myosin II-dependent contractile mechanism. Using the potent myosin II inhibitor blebbistatin, we demonstrate that traction forces are strongly dependent on a functional myosin II heavy chain. Since myosin II is regulated by both the myosin light chain kinase (NILCK) and, directly or indirectly, the Rho-associated kinase (ROCK), we examined the effects of inhibitors against these kinases. Interestingly, inhibition of the myosin light chain kinase had no detectable effect, while inhibition of the Rho-dependent kinase caused strong inhibition of traction forces. Our results indicate that ROCK and NILCK play non-redundant roles in regulating myosin II functions, and that a subset of myosin II, regulated by the Rho small GTPase, may be responsible for the regulation of traction forces in migrating fibroblasts. (c) 2006 Elsevier Inc. All rights reserved.
引用
收藏
页码:224 / 231
页数:8
相关论文
共 49 条
[1]   Different molecular motors mediate platelet-derived growth factor and lysophosphatidic acid-stimulated floating collagen matrix contraction [J].
Abe, M ;
Ho, CH ;
Kamm, KE ;
Grinnell, F .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (48) :47707-47712
[2]   The structural basis of blebbistatin inhibition and specificity for myosin II [J].
Allingham, JS ;
Smith, R ;
Rayment, I .
NATURE STRUCTURAL & MOLECULAR BIOLOGY, 2005, 12 (04) :378-379
[3]   Regulation and functions of Rho-associated kinase [J].
Amano, M ;
Fukata, Y ;
Kaibuchi, K .
EXPERIMENTAL CELL RESEARCH, 2000, 261 (01) :44-51
[4]   Phosphorylation and activation of myosin by Rho-associated kinase (Rho-kinase) [J].
Amano, M ;
Ito, M ;
Kimura, K ;
Fukata, Y ;
Chihara, K ;
Nakano, T ;
Matsuura, Y ;
Kaibuchi, K .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (34) :20246-20249
[5]   WORTMANNIN IS A POTENT PHOSPHATIDYLINOSITOL 3-KINASE INHIBITOR - THE ROLE OF PHOSPHATIDYLINOSITOL 3,4,5-TRISPHOSPHATE IN NEUTROPHIL RESPONSES [J].
ARCARO, A ;
WYMANN, MP .
BIOCHEMICAL JOURNAL, 1993, 296 :297-301
[6]   Flexible polyacrylamide substrata for the analysis of mechanical interactions at cell-substratum adhesions [J].
Beningo, KA ;
Lo, CM ;
Wang, YL .
METHODS IN CELL-MATRIX ADHESION, 2002, 69 :325-339
[7]   Flexible substrata for the detection of cellular traction forces [J].
Beningo, KA ;
Wang, YL .
TRENDS IN CELL BIOLOGY, 2002, 12 (02) :79-84
[8]   Nascent focal adhesions are responsible for the generation of strong propulsive forces in migrating fibroblasts [J].
Beningo, KA ;
Dembo, M ;
Kaverina, I ;
Small, JV ;
Wang, YL .
JOURNAL OF CELL BIOLOGY, 2001, 153 (04) :881-887
[9]   Adhesion-dependent cell mechanosensitivity [J].
Bershadsky, AD ;
Balaban, NQ ;
Geiger, B .
ANNUAL REVIEW OF CELL AND DEVELOPMENTAL BIOLOGY, 2003, 19 :677-695
[10]   A fluorescent resonant energy transfer-based biosensor reveals transient and regional myosin light chain kinase activation in lamella and cleavage furrows [J].
Chew, TL ;
Wolf, WA ;
Gallagher, PJ ;
Matsumura, F ;
Chisholm, RL .
JOURNAL OF CELL BIOLOGY, 2002, 156 (03) :543-553