Additive effect of Sp1 and Sp3 in regulation of the ED-L1E promoter of the EBV LMP 1 gene in human epithelial cells

The ED-L1E promoter of the LMP 1 gene is a GC box-containing promoter. To test if Sp1/Sp3 are important for modulating ED-L1E promoter activity through the GC box, site-specific mutation and deletion constructs carrying a reporter gene were transfected into NPCTW076 and C33A cells. Results showed that deletion or mutation of the GC box abolished ED-L1E activity. Association of Sp1/Sp3 with the GC box was confirmed by electrophoretic mobility shift and supershift assays using Spl- and Sp3-specific antibodies. Transfection of Spl- and Sp3-expressing vectors into NPCTRN076 and Sp- deficient Drosophila SL2 cells activated ED-L1E in a dose-dependent fashion and showed an additive effect. Data suggest that both factors may function as transcriptional activators and regulate the ED-L1E promoter in human epithelial cells.