FIG. 5.

EBNA1 recruits BGLF4 to oriP. (A) EMSA showing that EBNA1 DNA binding activity is not inhibited by BGLF4. EBNA1 nuclear extract (ext) was incubated with biotin-labeled DNA probe with (+) or without (−) preincubation with GST-BGLF4, as indicated. (B) ChIP assay examining the recruitment of BGLF4 to oriP. HeLa cells were transfected with an oriP-containing plasmid, pCEP4, and either V5-EBNA1c or V5-EBNA1c plus Flag-BGLF4. In cotransfected cells, pCEP4 oriP sequences were ChIPed by both anti-Flag (α-Flag) and α-V5 antibodies, indicating recruitment of Flag-BGLF4 to oriP in the presence of V5-EBNA1c. mIgG, control nonspecific mouse antibody. (C) ChIP assays performed on uninduced (left) and lytically induced (right) Akata cells. BGLF4 was recruited to oriP after lytic induction. mIgG served as the control antibody. FR and DS primer pairs, but not the BBLF1 and BOLF1 primer pairs, amplify the oriP region of the EBV genome.