Platelet-derived growth factor-induced H2O2 production requires the activation of phosphatidylinositol 3-kinase

Yun Soo Bae, Jee Young Sung, Ohn Soon Kim, Yeun Ju Kim, Kyu Chung Hur, Andrius Kazlauskas, Sue Goo Rhee

Research output: Contribution to journalArticlepeer-review

279 Scopus citations

Abstract

Autophosphorylation of the platelet-derived growth factor (PDGF) receptor triggers intracellular signaling cascades as a result of recruitment of Src homology 2 domain-containing enzymes, including phosphatidyl-inositol 3-kinase (PI3K), the GTPase-activating protein of Ras (GAP), the protein- tyrosine phosphatase SHP-2, and phospholipase C-γ1 (PLC-γ1), to specific phosphotyrosine residues. The roles of these various effectors in PDGF- induced generation of H2O2 have now been investigated in HepG2 cells expressing various PDGF receptor mutants. These mutants included a kinase- deficient receptor and receptors in which various combinations of the tyrosine residues required for the binding of PI3K (Tyr740 and Tyr751), GAP (Tyr771), SHP-2 (Tyr1009), or PLC-γ1 (Tyr1021) were mutated to Phe. PDGF failed to increase H2O2 production in cells expressing either the kinase-deficient mutant or a receptor in which the two Tyr residues required for the binding of PI3K were re- placed by Phe. In contrast, PDGF- induced H2O2 production in cells expressing a receptor in which the binding sites for GAP, SHP-2, and PLC-γ1 were all mutated was slightly greater than that in cells expressing the wild-type receptor. Only the PI3K binding site was alone sufficient for PDGF-induced H2O2 production. The effect of PDGF on H2O2 generation was blocked by the PI3K inhibitors LY294002 and wortmannin or by overexpression of a dominant negative mutant of Rac1. These results suggest that a product of PI3K is required for PDGF-induced production of H2O2 in nonphagocytic cells, and that Rac1 mediates signaling between the PI3K product and the putative NADPH oxidase.

Original languageEnglish
Pages (from-to)10527-10531
Number of pages5
JournalJournal of Biological Chemistry
Volume275
Issue number14
DOIs
StatePublished - 7 Apr 2000

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