Effect of EGF-receptor tyrosine kinase inhibitor on Rab5 function during endocytosis

Ivan Jozic, Samar C. Saliba, M. Alejandro Barbieri

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Tyrosine autophosphorylation within the cytoplasmic tail of EGF-receptor is a key event, which in turn recruits several factors including Shc, Grb2 and Rin1 that are essential activities for receptor-mediated endocytosis and signaling. In this study, we demonstrated that treatment with AG1478, an EGF-receptor kinase inhibitor, blocked the formation of Rab5-positive endosomes as well as the activation of Rab5 upon addition of EGF. We also found that EGF-receptor catalytically inactive mutant failed to activate Rab5 upon EGF stimulation. Additionally, endosomal co-localization of Rab5 and EGF-receptor was inhibited by AG1478. Interestingly, AG1478 inhibitor did not block the formation of enlarged Rab5-positive endosomes in cells expressing Rab5 GTP hydrolysis defective mutant (Rab5:Q79L). AG1478 inhibitor also blocked the in vitro endosome fusion in a concentration-dependent manner, and more importantly, Rab5:Q79L mutant rescued it. Furthermore, addition of Rin1, a Rab5 guanine nucleotide exchange factor, partially restored endosome fusion in the presence of AG1478 inhibitor. Consistent with these observations, we also observed that Rin1 was unable to localize to membranes upon EGF-stimulation in the presence of AG1478 inhibitor. These results constitute first evidence that the enzymatic activity of a tyrosine kinase receptor is required endosome fusion via the activation of Rab5.

Original languageEnglish (US)
Pages (from-to)16-24
Number of pages9
JournalArchives of Biochemistry and Biophysics
Volume525
Issue number1
DOIs
StatePublished - Sep 1 2012

Keywords

  • Endosome fusion
  • Kinase inhibitors
  • Receptor tyrosine kinase
  • Receptor-mediated encoytosis
  • Small GTPase

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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