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PMID: 17151359 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Regulation of myosin II dynamics by phosphorylation and dephosphorylation of its light chain in epithelial cells.

Molecular biology of the cell ·Vol. 18 ·No. 2 ·2007-02-00 ·Pages 605-16

Watanabe T, Hosoya H, Yonemura S

Abstract

Nonmuscle myosin II, an actin-based motor protein, plays an essential role in actin cytoskeleton organization and cellular motility. Although phosphorylation of its regulatory light chain (MRLC) is known to be involved in myosin II filament assembly and motor activity in vitro, it remains unclear exactly how MRLC phosphorylation regulates myosin II dynamics in vivo. We established clones of Madin Darby canine kidney II epithelial cells expressing MRLC-enhanced green fluorescent protein or its mutants. Time-lapse imaging revealed that both phosphorylation and dephosphorylation are required for proper dynamics of myosin II. Inhibitors affecting myosin phosphorylation and MRLC mutants indicated that monophosphorylation of MRLC is required and sufficient for maintenance of stress fibers. Diphosphorylated MRLC stabilized myosin II filaments and was distributed locally in regions of stress fibers where contraction occurs, suggesting that diphosphorylation is involved in the spatial regulation of myosin II assembly and contraction. We further found that myosin phosphatase or Zipper-interacting protein kinase localizes to stress fibers depending on the activity of myosin II ATPase.

MeSH Terms
Adenosine Triphosphatases/analysis,metabolism Animals Apoptosis Regulatory Proteins Calcium-Calmodulin-Dependent Protein Kinases Cells, Cultured Death-Associated Protein Kinases Dogs Epithelial Cells/chemistry,metabolism,ultrastructure Mutation Myosin Light Chains/analysis,genetics,metabolism Myosin Type II/analysis,genetics,metabolism Myosin-Light-Chain Phosphatase/analysis,metabolism Phosphorylation Protein Serine-Threonine Kinases/analysis,metabolism Stress Fibers/enzymology,metabolism,ultrastructure
Chemicals
Apoptosis Regulatory Proteins Myosin Light Chains Death-Associated Protein Kinases Protein Serine-Threonine Kinases Calcium-Calmodulin-Dependent Protein Kinases Myosin-Light-Chain Phosphatase Adenosine Triphosphatases Myosin Type II
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Watanabe Toshiyuki
RIKEN, Center for Developmental Biology, Kobe 650-0047, Japan.
Hosoya Hiroshi
Yonemura Shigenobu
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e06-07-0590
Published
2007-02-00
Epub
2006-00-06
Pages
605-16
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1783795
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.e06-07-0590 PMC Full Text View journal details
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