Role of actin-filament disassembly in lamellipodium protrusion in motile cells revealed using the drug jasplakinolide.
BACKGROUND
In motile cells, protrusion of the lamellipodium (a type of cell margin) requires assembly of actin monomers into actin filaments at the tip of the lamellipodium. The importance of actin-filament disassembly in this process is less well understood, and is assessed here using the actin drug jasplakinolide, which has two known activities - inhibition of filament disassembly and induction of an increase in actin polymer.


RESULTS
In cells the two activities of jasplakinolide were found to be separable; 1 microM jasplakinolide could permeate cells, bind cellular filamentous actin (F-actin) and inhibit filament disassembly within 3.5 minutes, but significant increase in actin polymer was not detected until 60 minutes of treatment. In live, permeabilised cells, jasplakinolide did not inhibit filament assembly from supplied, purified actin monomers. In migrating chick fibroblasts, lamellipodium protrusion was blocked within 1-5 minutes of treatment with 1 microM jasplakinolide, without any perturbation of actin organisation. In non-migrating chick fibroblasts, there was a delay in the onset of jasplakinolide-induced inhibition of lamellipodium protrusion, during which lamellipodium length increased linearly with no increase in protrusion rate. Motility of the bacterium Listeria in infected PtK2 cells was reduced 2.3-fold within 3 minutes of treatment with 1 microM jasplakinolide.


CONCLUSIONS
Actin-filament disassembly is tightly coupled to lamellipodium protrusion in migrating chick fibroblasts and motility of Listeria in PtK2 cells. One simple interpretation of these data is a situation whereby ongoing actin-filament assembly uses free actin monomer derived from filament disassembly, in preference to stored monomer.
Cramer
L P
LP
MRC-Laboratory Molecular Cell Biology University College London Gower Street, WC1E 6BT, The Randall Institute Kings College London, WC2B 5RL, UK. l.cramer@ucl.ac.uk.
D016475:3T3 Cells; D000199:Actins / Q000187:drug effects* / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D002642:Chick Embryo; D003599:Cytoskeleton / Q000187:drug effects; D047630:Depsipeptides; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects; D007700:Kinetics; D008089:Listeria monocytogenes / Q000166:cytology; D051379:Mice; D008856:Microscopy, Fluorescence; D010456:Peptides, Cyclic / Q000494:pharmacology; D013997:Time Factors
D000199:Actins; D047630:Depsipeptides; D010456:Peptides, Cyclic; C057531:jasplakinolide