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.     

Proliferation of both somatic and germ cells is affected in the Drosophila mutants of raf proto-oncogene.

The genomic and cDNA fragments of Drosophila melanogaster, homologous to human c-raf-1, were cloned. The nucleotide sequence predicted the primary structure of a polypeptide of 666 amino acid residues with a highly conserved Ser-Thr kinase domain on its carboxy terminal half. Draf-1 was mapped to the 2F region of the X chromosome. Two newly induced recessive lethals belonging to a complementation group in this region were identified to be defective in Draf-1 by P element-mediated rescue experiments. The mutants die at larval/pupal stages. The mutant larvae are apparently normal, but they harbor serious defects in the organs containing proliferating cells of both somatic and germ line origins. Maternal effects on embryogenesis indicated that Draf-1 is also required in early larval development.