Kroeger Paul T, Tokusumi Tsuyoshi, Schulz Robert A
Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA.
Genesis. 2012 Jan;50(1):41-9. doi: 10.1002/dvg.20787. Epub 2011 Oct 18.
Eater is a transmembrane protein that mediates phagocytosis in Drosophila. eater was identified in a microarray analysis of genes downregulated in S2 cells, in which Serpent had been knocked down by RNAi. The gene was shown to be expressed predominantly in plasmatocytes after embryonic development. We have extensively analyzed the transcriptional enhancer controlling eater expression with the following findings: the enhancer reproduces the plasmatocyte expression pattern of the gene as verified by anti-P1 antibody staining and a 526-basepair DNA region is active in lymph gland and hemolymph plasmatocytes. This DNA contains several GATA elements that serve as putative-binding sites for Serpent. Site-directed mutagenesis of two of these GATA sites abolishes eater expression in both lymph gland and hemolymph plasmatocytes. This suggests that Serpent regulates eater expression by binding these GATA sites, which was confirmed by gel shift analysis. These analyses allowed us to use eater-Gal4 to force plasmatocyte to lamellocyte differentiation.
Eater是一种跨膜蛋白,在果蝇中介导吞噬作用。eater是在对S2细胞中下调基因进行微阵列分析时发现的,在这些细胞中,Serpent已通过RNA干扰被敲低。该基因在胚胎发育后主要在浆血细胞中表达。我们广泛分析了控制eater表达的转录增强子,有以下发现:通过抗P1抗体染色验证,该增强子重现了该基因的浆血细胞表达模式,并且一个526碱基对的DNA区域在淋巴腺和血淋巴浆血细胞中具有活性。该DNA包含几个GATA元件,作为Serpent的假定结合位点。对其中两个GATA位点进行定点诱变可消除淋巴腺和血淋巴浆血细胞中的eater表达。这表明Serpent通过结合这些GATA位点来调节eater表达,凝胶迁移分析证实了这一点。这些分析使我们能够利用eater - Gal4促使浆血细胞向片状细胞分化。