Specific tyrosine phosphorylation sites on cortactin regulate Nck1-dependent actin polymerization in invadopodia

Matthew Oser, Christopher C. Mader, Hava Gil-Henn, Marco Magalhaes, Jose Javier Bravo-Cordero, Anthony J. Koleske, John S. Condeelis

Research output: Contribution to journalArticle

93 Citations (Scopus)

Abstract

Invadopodia are matrix-degrading membrane protrusions in invasive carcinoma cells enriched in proteins that regulate actin polymerization. The on-off regulatory switch that initiates actin polymerization in invadopodia requires phosphorylation of tyrosine residues 421, 466, and 482 on cortactin. However, it is unknown which of these cortactin tyrosine phosphorylation sites control actin polymerization. We investigated the contribution of individual tyrosine phosphorylation sites (421, 466, and 482) on cortactin to the regulation of actin polymerization in invadopodia. We provide evidence that the phosphorylation of tyrosines 421 and 466, but not 482, is required for the generation of free actin barbed ends in invadopodia. In addition, these same phosphotyrosines are important for Nck1 recruitment to invadopodia via its SH2 domain, for the direct binding of Nck1 to cortactin in vitro, and for the FRET interaction between Nck1 and cortactin in invadopodia. Furthermore, matrix proteolysis-dependent tumor cell invasion is dramatically inhibited in cells expressing a mutation in phosphotyrosine 421 or 466. Together, these results identify phosphorylation of tyrosines 421 and 466 on cortactin as the crucial residues that regulate Nck1-dependent actin polymerization in invadopodia and tumor cell invasion, and suggest that specifically blocking either tyrosine 421 or 466 phosphorylation might be effective at inhibiting tumor cell invasion in vivo.

Original languageEnglish (US)
Pages (from-to)3662-3673
Number of pages12
JournalJournal of Cell Science
Volume123
Issue number21
DOIs
StatePublished - Nov 1 2010

Fingerprint

Cortactin
Polymerization
Tyrosine
Actins
Phosphorylation
Phosphotyrosine
Neoplasms
src Homology Domains
Podosomes
Proteolysis
Carcinoma
Mutation
Membranes

Keywords

  • Cancer invasion
  • Cortactin
  • Invadopodia
  • Nck1
  • Phosphorylation
  • SH2

ASJC Scopus subject areas

  • Cell Biology

Cite this

Specific tyrosine phosphorylation sites on cortactin regulate Nck1-dependent actin polymerization in invadopodia. / Oser, Matthew; Mader, Christopher C.; Gil-Henn, Hava; Magalhaes, Marco; Bravo-Cordero, Jose Javier; Koleske, Anthony J.; Condeelis, John S.

In: Journal of Cell Science, Vol. 123, No. 21, 01.11.2010, p. 3662-3673.

Research output: Contribution to journalArticle

Oser M, Mader CC, Gil-Henn H, Magalhaes M, Bravo-Cordero JJ, Koleske AJ et al. Specific tyrosine phosphorylation sites on cortactin regulate Nck1-dependent actin polymerization in invadopodia. Journal of Cell Science. 2010 Nov 1;123(21):3662-3673. https://doi.org/10.1242/jcs.068163
Oser, Matthew ; Mader, Christopher C. ; Gil-Henn, Hava ; Magalhaes, Marco ; Bravo-Cordero, Jose Javier ; Koleske, Anthony J. ; Condeelis, John S. / Specific tyrosine phosphorylation sites on cortactin regulate Nck1-dependent actin polymerization in invadopodia. In: Journal of Cell Science. 2010 ; Vol. 123, No. 21. pp. 3662-3673.
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