Transcriptional regulation of the Drosophila raf proto-oncogene by Drosophila STAT during development and in immune response.
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.
Kwon
E J
EJ
Laboratory of Cell Biology, Aichi Cancer Center Research Institute, Chikusa-ku, Nagoya, 464-8681, Japan.
Park
H S
HS
Kim
Y S
YS
Oh
E J
EJ
Nishida
Y
Y
Matsukage
A
A
Yoo
M A
MA
Yamaguchi
M
M
D000818:Animals; D030801:Animals, Genetically Modified; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004330:Drosophila / Q000235:genetics* / Q000276:immunology; D029721:Drosophila Proteins; D005260:Female; D005697:Galactosides / Q000378:metabolism; D005786:Gene Expression Regulation; D018507:Gene Expression Regulation, Developmental; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007211:Indoles / Q000378:metabolism; D053612:Janus Kinases; D008156:Luciferases / Q000378:metabolism; D020935:MAP Kinase Signaling System; D008297:Male; D008957:Models, Genetic; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D019908:Proto-Oncogene Proteins c-raf / Q000235:genetics / Q000378:metabolism*; D012045:Regulatory Sequences, Nucleic Acid; D013997:Time Factors; D014157:Transcription Factors; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014162:Transfection; D014466:Ultraviolet Rays; D014638:Vanadates / Q000494:pharmacology; D001616:beta-Galactosidase / Q000378:metabolism
D029721:Drosophila Proteins; D005697:Galactosides; D007211:Indoles; D014157:Transcription Factors; D014638:Vanadates; D006861:Hydrogen Peroxide; D008156:Luciferases; D011505:Protein-Tyrosine Kinases; D053612:Janus Kinases; C085845:hop protein, Drosophila; D019908:Proto-Oncogene Proteins c-raf; D001616:beta-Galactosidase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside