Skip to main content
. 1998 Apr;72(4):3146–3154. doi: 10.1128/jvi.72.4.3146-3154.1998

FIG. 8.

FIG. 8

(A) In vitro transcription templates. p5BSE4CAT contains five p53 binding sites upstream of the Ad E4 TATA box and CAT gene. This template is responsive to activation by p53. The primer extensions of RNAs transcribed from this template are 160 nucleotides (160 n) in length. pG5E1BCAT contains five Gal4 binding sites upstream of the Ad E1B TATA box and CAT gene. This template is responsive to activation by Gal4-AH. Primer extensions of RNAs transcribed from this template are 110 nucleotides (110 n) in length. (B) e55K specifically represses p53-mediated activation at the transcriptional level. In vitro transcription reaction mixtures contained HeLa nuclear extract, template DNAs, and ep53 (20 ng [lanes 3 to 8 and 11 to 16]), Gal4-AH (10 ng [lanes 2, 4 to 8, and 10, and 12 to 16]) and increasing concentrations of e55K (10 ng/μl [lanes 5 to 8, 2, 4, 8, and 16 μl, respectively]) or increasing concentrations of the control BP6 protein fraction (lanes 13 to 16, 2, 4, 8, and 16 μl, respectively). Primer extension products (160 n and 110 n) indicate levels of transcription arising from pB5E4CAT and pG5E1BCAT, respectively and their positions are indicated by arrows. (C) In vitro transcription with templates p5BSCAT and pG5E1BCAT; HeLa nuclear extract; and 40 ng of ep53 (lanes 2 to 5); 10 ng of Gal4-AH; and 30, 70, and 140 ng of e55K (lanes 3, 4, and 5, respectively).