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PMID: 14757754 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Distinct roles of MLCK and ROCK in the regulation of membrane protrusions and focal adhesion dynamics during cell migration of fibroblasts.

The Journal of cell biology ·Vol. 164 ·No. 3 ·2004-02-02 ·Pages 427-39

Totsukawa G, Wu Y, Sasaki Y, Hartshorne DJ, Yamakita Y, Yamashiro S, Matsumura F

Abstract

We examined the role of regulatory myosin light chain (MLC) phosphorylation of myosin II in cell migration of fibroblasts. Myosin light chain kinase (MLCK) inhibition blocked MLC phosphorylation at the cell periphery, but not in the center. MLCK-inhibited cells did not assemble zyxin-containing adhesions at the periphery, but maintained focal adhesions in the center. They generated membrane protrusions all around the cell, turned more frequently, and migrated less effectively. In contrast, Rho-associated kinase (ROCK) inhibition blocked MLC phosphorylation in the center, but not at the periphery. ROCK-inhibited cells assembled zyxin-containing adhesions at the periphery, but not focal adhesions in the center. They moved faster and more straight. On the other hand, inhibition of myosin phosphatase increased MLC phosphorylation and blocked peripheral membrane ruffling, as well as turnover of focal adhesions and cell migration. Our results suggest that myosin II activated by MLCK at the cell periphery controls membrane ruffling, and that the spatial regulation of MLC phosphorylation plays critical roles in controlling cell migration of fibroblasts.

MeSH Terms
Actins/metabolism Animals Cell Adhesion/physiology Cell Movement/physiology Cell Size Cell Surface Extensions/metabolism Cells, Cultured Enzyme Inhibitors/metabolism Fibroblasts/cytology,metabolism Fibroma Focal Adhesions/metabolism Gerbillinae Humans Intracellular Signaling Peptides and Proteins Microscopy, Video Myosin-Light-Chain Kinase/metabolism Myosin-Light-Chain Phosphatase/antagonists & inhibitors,metabolism Phosphorylation Protein Serine-Threonine Kinases/metabolism rho-Associated Kinases
Chemicals
Actins Enzyme Inhibitors Intracellular Signaling Peptides and Proteins Protein Serine-Threonine Kinases rho-Associated Kinases Myosin-Light-Chain Kinase Myosin-Light-Chain Phosphatase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Totsukawa Go
Department of Molecular Biology and Biochemistry, Rutgers University, Nelson Labs, Rm. A323, 604 Allison Rd., Piscataway, NJ 08855, USA.
Wu Yue
Sasaki Yasuharu
Hartshorne David J
Yamakita Yoshihiko
Yamashiro Shigeko
Matsumura Fumio
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200306172
Published
2004-02-02
Pages
427-39
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2172229
Subset
IM
Grants
NHLBI NIH HHS · HL23615 · United States
NCI NIH HHS · R37 CA042742 · United States
NCI NIH HHS · R01 CA042742 · United States
NCI NIH HHS · CA42742 · United States
NHLBI NIH HHS · R01 HL023615 · United States
External Links
PubMed source DOI: 10.1083/jcb.200306172 PMC Full Text View journal details
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