EGF stimulates an increase in actin nucleation and filament number at the leading edge of the lamellipod in mammary adenocarcinoma cells

Amanda Y. Chan, Steven Raft, Maryse Bailly, Jeffrey B. Wyckoff, Jeffrey E. Segall, John S. Condeelis

Research output: Contribution to journalArticle

177 Citations (Scopus)

Abstract

Stimulation of metastatic MTLn3 cells with EGF causes the rapid extension of lamellipods, which contain a zone of F-actin at the leading edge. In order to establish the mechanism for accumulation of F-actin at the leading edge and its relationship to lamellipod extension in response to EGF, we have studied the kinetics and location of EGF-induced actin nucleation activity in MTLn3 cells and characterized the actin dynamics at the leading edge by measuring the changes at the pointed and barbed ends of actin filaments upon EGF stimulation of MTLn3 cells. The major result of this study is that stimulation of MTLn3 cells with EGF causes a transient increase in actin nucleation activity resulting from the appearance of free barbed ends very close to the leading edge of extending lamellipods. In addition, cytochalasin D causes a significant decrease in the total F-actin content in EGF-stimulated cells, indicating that both actin polymerization and depolymerization are stimulated by EGF. Pointed end incorporation of rhodamine-labeled actin by the EGF stimulated cells is 2.12 ± 0.47 times higher than that of control cells. Since EGF stimulation causes an increase in both barbed and pointed end incorporation of rhodamine-labeled actin in the same location, the EGF-stimulated nucleation sites are more likely due either to severing of pre-existing filaments or de novo nucleation of filaments at the leading edge thereby creating new barbed and pointed ends. The timing and location of EGF-induced actin nucleation activity in MTLn3 cells can account for the observed accumulation of F-actin at the leading edge and demonstrate that this F-actin rich zone is the primary actin polymerization zone after stimulation.

Original languageEnglish (US)
Pages (from-to)199-211
Number of pages13
JournalJournal of Cell Science
Volume111
Issue number2
StatePublished - 1998

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Actin Cytoskeleton
Epidermal Growth Factor
Actins
Adenocarcinoma
Breast
Rhodamines
Polymerization
Cytochalasin D

Keywords

  • Actin nucleation
  • EGF
  • Metastasis
  • Motility

ASJC Scopus subject areas

  • Cell Biology

Cite this

EGF stimulates an increase in actin nucleation and filament number at the leading edge of the lamellipod in mammary adenocarcinoma cells. / Chan, Amanda Y.; Raft, Steven; Bailly, Maryse; Wyckoff, Jeffrey B.; Segall, Jeffrey E.; Condeelis, John S.

In: Journal of Cell Science, Vol. 111, No. 2, 1998, p. 199-211.

Research output: Contribution to journalArticle

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AU - Segall, Jeffrey E.

AU - Condeelis, John S.

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