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PMID:10764759

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Citation

Kwon, EJ, Park, HS, Kim, YS, Oh, EJ, Nishida, Y, Matsukage, A, Yoo, MA and Yamaguchi, M (2000) Transcriptional regulation of the Drosophila raf proto-oncogene by Drosophila STAT during development and in immune response. J. Biol. Chem. 275:19824-30

Abstract

The Drosophila raf (D-raf) gene promoter contains a recognition consensus sequence for Drosophila STAT (D-STAT). By band mobility shift assay, we detected a factor binding to the D-STAT-recognition sequence in extracts of cultured Drosophila cells treated with vanadate peroxide. UV-cross-linking analyses suggested the size of the binding factor to be almost same as that of D-STAT. Furthermore, the binding activity was increased in cells cotransfected with HOP and D-STAT expression plasmids. These results strongly suggest that D-STAT binds to the D-STAT recognition sequence in the D-raf gene promoter. Transient luciferase expression assay using Schneider 2 cells indicated that the D-raf gene promoter is activated by D-STAT through the D-STAT-binding site. Furthermore, analyses with transgenic flies carrying Draf-lacZ fusion genes with and without mutations in the D-STAT-binding site pointed to an important role in D-raf gene promoter activity throughout development. We also found that the D-STAT-binding site is required for injury-induced activation of the D-raf gene promoter. Here we propose that D-STAT can participate in regulation of the mitogen-activated protein kinase cascade through D-raf gene activation.

Links

PubMed Online version:10.1074/jbc.M001114200

Keywords

Animals; Animals, Genetically Modified; Cells, Cultured; Dose-Response Relationship, Drug; Drosophila/genetics; Drosophila/immunology; Drosophila Proteins; Female; Galactosides/metabolism; Gene Expression Regulation; Gene Expression Regulation, Developmental; Hydrogen Peroxide/pharmacology; Indoles/metabolism; Janus Kinases; Luciferases/metabolism; MAP Kinase Signaling System; Male; Models, Genetic; Plasmids; Promoter Regions, Genetic; Protein Binding; Protein-Tyrosine Kinases/metabolism; Proto-Oncogene Proteins c-raf/genetics; Proto-Oncogene Proteins c-raf/metabolism; Regulatory Sequences, Nucleic Acid; Time Factors; Transcription Factors; Transcription, Genetic; Transcriptional Activation; Transfection; Ultraviolet Rays; Vanadates/pharmacology; beta-Galactosidase/metabolism

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