Transcriptional regulation of eater gene expression in Drosophila blood cells

• Published in: Genesis (New York, N.Y. : 2000) N.Y. : 2000), 2012-01, Vol.50 (1), p.41-49
• Main Authors: Kroeger Jr, Paul T., Tokusumi, Tsuyoshi, Schulz, Robert A.
• Format: Article
• Language: English
• Subjects: Animals; Binding Sites - genetics; Blood Cells - metabolism; Cell Differentiation; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Drosophila; Drosophila - cytology; Drosophila - embryology; Drosophila - genetics; Drosophila Proteins - genetics; Drosophila Proteins - metabolism; Electrophoretic Mobility Shift Assay - methods; enhancer analysis; Enhancer Elements, Genetic; GATA Transcription Factors - genetics; GATA Transcription Factors - metabolism; Gene Expression Regulation, Developmental; Hematopoiesis; Hemocytes - cytology; Hemocytes - metabolism; Membrane Proteins - genetics; Membrane Proteins - metabolism; Multigene Family; Mutagenesis, Site-Directed; plasmatocyte; Receptors, Cell Surface - genetics; Receptors, Cell Surface - metabolism; RNA Interference; Serpent; Transcription, Genetic
• ISSN: 1526-954X; 1526-968X
• DOI: 10.1002/dvg.20787

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. genesis 50:41–49, 2012. © 2011 Wiley Periodicals, Inc.