A fluorescent resonant energy transfer-based biosensor reveals transient and regional myosin light chain kinase activation in lamella and cleavage furrows

Teng Leong Chew, Wendy A. Wolf, Patricia J. Gallagher, Fumio Matsumura, Rex L. Chisholm

Research output: Contribution to journalArticle

100 Citations (Scopus)

Abstract

Approaches with high spatial and temporal resolution are required to understand the regulation of nonmuscle myosin II in vivo. Using fluorescence resonance energy transfer we have produced a novel biosensor allowing simultaneous determination of myosin light chain kinase (MLCK) localization and its [Ca2+]4/calmodulin-binding state in living cells. We observe transient recruitment of diffuse MLCK to stress fibers and its in situ activation before contraction. MLCK is highly active in the lamella of migrating cells, but not at the retracting tail. This unexpected result highlights a potential role for MLCK-mediated myosin contractility in the lamella as a driving force for migration. During cytokinesis, MLCK was enriched at the spindle equator during late metaphase, and was maximally activated just before cleavage furrow constriction. As furrow contraction was completed, active MLCK was redistributed to the poles of the daughter cells. These results show MLCK is a myosin regulator in the lamella and contractile ring, and pinpoints sites where myosin function may be mediated by other kinases.

Original languageEnglish (US)
Pages (from-to)543-553
Number of pages11
JournalJournal of Cell Biology
Volume156
Issue number3
DOIs
StatePublished - Feb 4 2002

Fingerprint

Myosin-Light-Chain Kinase
Energy Transfer
Biosensing Techniques
Myosins
Myosin Type II
Stress Fibers
Fluorescence Resonance Energy Transfer
Cytokinesis
Calmodulin
Metaphase
Constriction
Phosphotransferases

Keywords

  • Cell division
  • FRET
  • Myosin light chain kinase
  • Myosin light chains
  • Phosphorylation

ASJC Scopus subject areas

  • Cell Biology

Cite this

A fluorescent resonant energy transfer-based biosensor reveals transient and regional myosin light chain kinase activation in lamella and cleavage furrows. / Chew, Teng Leong; Wolf, Wendy A.; Gallagher, Patricia J.; Matsumura, Fumio; Chisholm, Rex L.

In: Journal of Cell Biology, Vol. 156, No. 3, 04.02.2002, p. 543-553.

Research output: Contribution to journalArticle

Chew, Teng Leong ; Wolf, Wendy A. ; Gallagher, Patricia J. ; Matsumura, Fumio ; Chisholm, Rex L. / A fluorescent resonant energy transfer-based biosensor reveals transient and regional myosin light chain kinase activation in lamella and cleavage furrows. In: Journal of Cell Biology. 2002 ; Vol. 156, No. 3. pp. 543-553.
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