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. 2004 Sep;78(17):9431–9445. doi: 10.1128/JVI.78.17.9431-9445.2004

FIG. 7.

FIG. 7.

Analysis of the effects of zipper and basic region mutations on the ability of EBNA 3C to repress EBNA 2 activation of the C promoter in vivo. (A) Amino acid sequence of the EBNA 3C bZIP region, showing the locations of the mutations introduced. (B) Effect of leucine-to-proline substitutions in the zipper region on the repression activity of EBNA 3C. DG75 cells were transiently transfected with the indicated amounts of plasmids pSG52A and pSG5EBNA3C in addition to 2 μg of C promoter reporter (pCp1425-GL2) and 2 μg of the control plasmid pRL-CMV. The Renilla luciferase activity (pRL-CMV) was used to correct for transfection efficiency. Results are expressed as transactivation by EBNA 2 relative to the level of transcription obtained in the absence of EBNA 2 and EBNA 3C. The bar charts show the mean ± standard deviation of three independent experiments. Samples of cell lysates (adjusted for transfection efficiency) were separated by SDS-PAGE and Western blotted, and EBNA 3C expression was detected with the EBNA 3C monoclonal antibody E3CA10. (C) Effect of basic domain mutations on the repression activity of EBNA 3C. (D) Effect of mutating the RBP-Jκ binding site (TFGC to AAAA at positions 209 to 212) on the repression activity of EBNA 3C.