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1 Venture Laboratory, Kyoto Institute of Technology, Sakyo-ku, Kyoto, Japan;
2 Department of Applied Biology, Kyoto Institute of Technology, Sakyo-ku, Kyoto, Japan;
3 Division of Biochemistry, Aichi Cancer Center Research Institute, Chikusa-ku, Nagoya, Japan
The DNA replication-related element binding factor (DREF) has been suggested as being involved in regulation of DNA replication- and proliferation-related genes in Drosophila. Recently, by searching the Drosophila genome database, we also found DRE-like sequences in the 5'-flanking regions of many genes with other functions. In addition, immunostaining of polytene chromosomes with an anti-DREF monoclonal antibody revealed that DREF can bind to a hundred regions of polytene chromosomes, suggesting regulation of multiple genes and multiple roles in vivo. When we over-expressed DREF protein or inverted repeat RNA of the DREF gene in wing imaginal discs using the GAL4-UAS targeted expression system in Drosophila, the results were veins of increased width and a loss of veins, respectively. With DREF over-expression, Rolled, a Drosophila MAPK homologue, was ectopically activated. Furthermore, half reduction of the D-raf gene dose suppressed this DREF-induced vein of increased width phenotype. In addition, when DREF transcripts were reduced by introducing double-stranded RNA of the DREF gene into S2 cells, the D-raf gene promoter activity was diminished to 4%. These data indicate that DREF is involved in regulation of vein formation through the activation of EGFR signalling in the Drosophila wing imaginal discs.
Present addresses:aLaboratory of Cell Biology, Department of Bioinformatics, Faculty of Engineering, Soka University, 1-236 Tangi-cho, Hachioji, Tokyo 192-8577, Japan; bLaboratory of Molecular Oncology, MGH Cancer Center Building 149, 13th Street, Charlestown, MA 02129-2000, USA; cDepartment of Life Science, Graduate School of Science, Himeji Institute of Technology, 3-2-1 Koto, Kamigori, Hyogo 678-1297, Japan. * Correspondence: Email: myamaguc{at}kit.ac.jp
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