Rapid actin transport during cell protrusion

Science. 2003 Apr 4;300(5616):142-5. doi: 10.1126/science.1082026.

Abstract

Transformed rat fibroblasts expressing two variants of green fluorescent protein, each fused to beta-actin, were used to study actin dynamics during cell protrusion. The recently developed FLAP (fluorescence localization after photobleaching) method permits the tracking of one fluorophore after localized photobleaching by using the other as a colocalized reference. Here, by visualizing the ratio of bleached to total molecules, we found that actin was delivered to protruding zones of the leading edge of the cell at speeds that exceeded 5 micrometers per second. Monte Carlo modeling confirmed that this flow cannot be explained by diffusion and may involve active transport.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism*
  • Amides / pharmacology
  • Animals
  • Azepines / pharmacology
  • Bacterial Proteins / metabolism
  • Biological Transport, Active
  • Biopolymers
  • Cell Line, Transformed
  • Cell Movement
  • Depsipeptides*
  • Diffusion
  • Enzyme Inhibitors / pharmacology
  • Fluorescence
  • Fluorescence Recovery After Photobleaching
  • Fluorometry
  • Green Fluorescent Proteins
  • Image Processing, Computer-Assisted
  • Intracellular Signaling Peptides and Proteins
  • Luminescent Proteins / metabolism
  • Microscopy, Confocal
  • Monte Carlo Method
  • Myosin-Light-Chain Kinase / antagonists & inhibitors
  • Myosin-Light-Chain Kinase / metabolism
  • Naphthalenes / pharmacology
  • Nocodazole / pharmacology
  • Peptides, Cyclic / pharmacology
  • Photobleaching
  • Protein Serine-Threonine Kinases / antagonists & inhibitors
  • Protein Serine-Threonine Kinases / metabolism
  • Protein Transport / drug effects
  • Pseudopodia / drug effects
  • Pseudopodia / physiology*
  • Pseudopodia / ultrastructure*
  • Pyridines / pharmacology
  • Rats
  • Recombinant Fusion Proteins / metabolism
  • Tumor Cells, Cultured
  • rho-Associated Kinases

Substances

  • Actins
  • Amides
  • Azepines
  • Bacterial Proteins
  • Biopolymers
  • Depsipeptides
  • Enzyme Inhibitors
  • Intracellular Signaling Peptides and Proteins
  • Luminescent Proteins
  • Naphthalenes
  • Peptides, Cyclic
  • Pyridines
  • Recombinant Fusion Proteins
  • yellow fluorescent protein, Bacteria
  • jasplakinolide
  • ML 7
  • Y 27632
  • Green Fluorescent Proteins
  • Protein Serine-Threonine Kinases
  • rho-Associated Kinases
  • Myosin-Light-Chain Kinase
  • Nocodazole