FIG. 4.

Analysis of ORF70-K4 intergenic region. (A) Full-length (FL) and truncated (Δ1, Δ2, Δ3, Δ4, and Δ5) versions of ORF70hp-luc were tested for a response to RTA in HeLa cells by transient transfection. Assays were performed in triplicate and the luciferase values expressed as the mean and standard deviation. For each reporter, fold induction was calculated from luciferase activity measured in the presence and absence of the RTA expression plasmid. (B) Annotated sequence of the ORF70hp fragment (nucleotides 21,105 to 21,842 in the M-type reference sequence). Overlap with ORFK4 (encoding vMIP2) is shown in lowercase, as are the first few codons of ORF70, encoding thymidylate synthase. Also shown are the end points of the truncations used in panel A (filled flags), the four candidate CSL sites, ORF70.1, ORF70.2, ORF70.3, and ORF70.4 (filled boxes), and TSSs mapped by 5′-RACE (filled or open lollipops). The principal TSS (filled lollipop) is labeled +1 and corresponds to position 21,150. A putative TATA box and consensus NF-κB site are indicated with an open box.