Title

Ligand-independent phosphorylation of Y869 (Y845) links mutant EGFR signaling to stat-mediated gene expression

Authors

Authors

S. Yanga; K. Parka; J. Turkson;C. L. Arteaga

Comments

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Abbreviated Journal Title

Exp. Cell Res.

Keywords

EGFR; Src; lung cancer; STAT; phosphorylation; GROWTH-FACTOR RECEPTOR; TYROSINE KINASE INHIBITOR; CELL LUNG-CANCER; C-SRC; INCREASED SENSITIVITY; DOMAIN MUTATIONS; DOWN-REGULATION; GEFITINIB; PROTEINS; TRANSFORMATION; Oncology; Cell Biology

Abstract

Activating mutants of EGFR have been identified in a subset of non-small-cell lung cancers. To investigate mutant-driven signaling, we focused on Y869, a residue in the same activation loop where the L858R and L861Q mutations are located. We observed ligand-independent phosphorylation of Y869 in 32D cells EGFR(L858R) and EGFR(L861Q). The EGFR tyrosine kinase inhibitor (TKI) erlotinib inhibited Y869 P-EGFR in intact cells as well as in a cell-free kinase reaction. Expression of kinase domain of EGFR(L858R) and EGFR(L861Q) exhibited auto-phosphorylation of Y869; this was inhibited by EGFR TKIs but not by Src kinase inhibitor. P-Y859 of EGFR-mediated downstream component, STAT5, was also analyzed. Y694 P-STAT5 was eliminated by erlotinib treatment. Analysis of immune-complexes showed constitutive association of mutant EGFRs with STAT5 and Src which was unaffected by erlotinib or PP1. On the other hand, 32D-EGFR(WT) exhibited constitutive STAT5 phosphorylation and association of EGFR with JAK2. In these cells, a JAK2 inhibitor abrogated P-STAT5 whereas mutant EGFRs did not associate with JAK2. Expression of c-myc was regulated by EGFR/STAT5 signaling in cells expressing EGFR(L858R) and EGFR(L861Q). Our results suggest that ligand-independent and Src activity-independent phosphorylation of Y869 in mutant EGFR regulates STAT5 activation and c-myc expression. (C) 2007 Elsevier Inc. All rights reserved.

Journal Title

Experimental Cell Research

Volume

314

Issue/Number

2

Publication Date

1-1-2008

Document Type

Article

Language

English

First Page

413

Last Page

419

WOS Identifier

WOS:000251891300018

ISSN

0014-4827

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