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

Go Totsukawa, Yue Wu, Yasuharu Sasaki, David J. Hartshorne, Yoshihiko Yamakita, Shigeko Yamashiro, Fumio Matsumura

Research output: Contribution to journalArticle

291 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)427-439
Number of pages13
JournalJournal of Cell Biology
Volume164
Issue number3
DOIs
StatePublished - Feb 2 2004

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Myosin-Light-Chain Kinase
rho-Associated Kinases
Myosin Light Chains
Focal Adhesions
Cell Movement
Fibroblasts
Phosphorylation
Zyxin
Membranes
Myosin Type II
Myosin-Light-Chain Phosphatase

All Science Journal Classification (ASJC) codes

  • Cell Biology

Keywords

  • Cell migration
  • Cell polarity
  • MLCK
  • Membrane protrusion
  • Myosin phosphorylation

Cite this

Totsukawa, Go ; Wu, Yue ; Sasaki, Yasuharu ; Hartshorne, David J. ; Yamakita, Yoshihiko ; Yamashiro, Shigeko ; Matsumura, Fumio. / Distinct roles of MLCK and ROCK in the regulation of membrane protrusions and focal adhesion dynamics during cell migration of fibroblasts. In: Journal of Cell Biology. 2004 ; Vol. 164, No. 3. pp. 427-439.
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Distinct roles of MLCK and ROCK in the regulation of membrane protrusions and focal adhesion dynamics during cell migration of fibroblasts. / Totsukawa, Go; Wu, Yue; Sasaki, Yasuharu; Hartshorne, David J.; Yamakita, Yoshihiko; Yamashiro, Shigeko; Matsumura, Fumio.

In: Journal of Cell Biology, Vol. 164, No. 3, 02.02.2004, p. 427-439.

Research output: Contribution to journalArticle

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AU - Totsukawa, Go

AU - Wu, Yue

AU - Sasaki, Yasuharu

AU - Hartshorne, David J.

AU - Yamakita, Yoshihiko

AU - Yamashiro, Shigeko

AU - Matsumura, Fumio

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N2 - 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.

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