v-Src accelerates spontaneous motility via phosphoinositide 3-kinase, phospholipase C and phospholipase D, but abrogates chemotaxis in Rat-1 and MDCK cells

Anna Platek, Marcel Mettlen, Isabelle Camby, Robert Kiss, Mustapha Amyere, Pierre J. Courtoy

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

In Rat-1 fibroblasts, v-Src causes a profound remodelling of cortical actin cytoskeleton. This transformation includes membrane ruffling, a hallmark of the leading edge in migrating cells, and results from activation of phosphoinositide 3-kinase (PI 3-kinase), phospholipase C (PLC) and phospholipase D (PLD). We therefore re-exammed whether motility is constitutively triggered by v-Src and studied whether this response is controlled by the same signalling pathway. The study was performed using Rat-1/tsLA29 and AMCK/tsLA31 cells, each harbouring a different thermosensitive v-Sre kinase, active at 34°C but inactivated at 40°C. In both cell lines, overnight v-Src activation induced transformation and accelerated spontaneous motility by approximately twofold, as evidenced by wound-healing assay and by single-cell track, time-lapse recording in Dunn chambers. Inhibitors of PI 3-kinase, PLC and PLD selectively abrogated acceleration of motility by v-Src. Since mechanisms that co-ordinate spontaneous, as distinct from oriented, cell migration are separable, we further analysed in Dunn chambers chemotactic response of Rat-1/tsLA29 cells to PDGF and of NMCK /tsLA31 cells to EGF. In both cases, v-Src decreased the steady-state level of growth factor receptors at the cell surface twofold, and abrogated movement directionality at comparable level of occupancy as in non-transformed cells. The burst of pinocytosis in response to growth factors was also abolished by v-Src. Altogether, these results indicate that v-Src triggers motility in a PI 3-kinase-, PLC-and PLD-dependent manner, but abrogates directionality by suppressing polarised signalling downstream of Prowth factor receptors.

Original languageEnglish (US)
Pages (from-to)4849-4861
Number of pages13
JournalJournal of cell science
Volume117
Issue number20
DOIs
StatePublished - Sep 15 2004

Keywords

  • Cell migration
  • Chemotaxis
  • PI 3-kinase
  • PLC
  • PLD
  • v-Src

ASJC Scopus subject areas

  • Cell Biology

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