Rapid Phosphorylation of Ets-2 Accompanies Mitogen-Activated Protein Kinase Activation and the Induction of Heparin-Binding Epidermal Growth Factor Gene Expression by Oncogenic Raf-1

• DOI: 10.1128/mcb.17.5.2401
• Publication Date: 2015-10-09T22:07:41Z
• Authors: Sean A. McCarthy; Dan Chen; Beom-Seok Yang; Jose J. Garcia Ra...; Holly Cherwinski; Xiao-Rong Chen; Michael Klagsbrun; Craig A. Hauser; Michael C. Ostrowski; Martin McMahon
• Keywords: c-Raf

Heparin-binding epidermal growth factor (HB-EGF) gene transcription is rapidly activated in NIH 3T3 cells transformed by oncogenic Ras and Raf mediates the autocrine activation of c-Jun N-terminal kinases (JNKs) observed these cells.A 1.7-kb fragment promoter murine HB-EGF linked to a luciferase reporter was strongly induced following ⌬Raf-1:ER, conditionally active form human Raf-1.Promoter ⌬Raf-1:ER required composite AP-1/Ets binding site located between bp ؊974 ؊988 upstream translation initiation site.In vivo genomic footprinting indicated that basal level occupancy this element increased activation.Cotransfection Ets-2 p44 mitogen-activated protein (MAP) kinase expression vectors potentiated response ⌬Raf-1:ER.Potentiated both MAP catalytic activity threonine 72 Pointed domain Ets-2.Biochemical assays demonstrated ability p42 phosphorylate on 72.Importantly, intact cells, kinetics phosphorylation residue closely mirror onset activation.These data firmly establish as direct target Raf-MEK-MAP signaling pathway implicate regulation expression.