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. 2002 Aug 1;16(15):1977–1989. doi: 10.1101/gad.996502

Figure 3.

Figure 3

Figure 3

Figure 3

RTA/RBP-Jκ interactions in authentic viral promoters. (A) Promoters of MTA (ORF57) and SSB contain RBPJκ recognition sites. The sequences of MTA and SSB promoters are shown, with the RBPJκ recognition site outlined by an empty box (orientation: reverse in MTA promoter and forward in SSB promoter). The previously identified 52-bp RTA-response element (50RE) between −106 and −54 nt of MTA promoter is underlined. (B) RTA transactivation of MTA or SSB promoters (WT or RBPJκ site-mutated) in OT13 (WT) or OT11 (RBPJκ−/−) cell line. OT13 or OT11 cells were cotransfected with 0 to 0.5 μg of pcDNA3-Flc50 and 0.5 μg of luciferase reporter constructs driven by MTA promoter, MTA promoter with mutations at RBPJκ recognition site (see Materials and Methods) (MTAmut), SSB promoter, or SSB promoter with mutations at RBPJκ recognition site (SSBmut). In OT11 cells, RTA transactivation was tested in the absence (−) or presence (+) of 1 μg of cotransfected pcDNA3.1-RBPJκ. (C) Schematic representation of mutants of PAN promoter and their responsiveness to RTA transactivation in SLK and OT11 cells. The RTA-response element (50RE) identified by Song et al. (2001) extends from −69 to −38 bp upstream of RNA start site. Mutation on 50RE, represented by a hatched box, was created by replacing the 12-nt sequence (−67 to −56) with linker sequence TATCATATGATA. RBPJκ recognition site was from −818 to −824 in reverse direction. Mutation of RBPJκ recognition site (changes of TTCCCACGG to TTCCAAGCC) is represented by the gray box. RTA transactivation on the WT and mutant PAN promoters in SLK and OT11 cells was measured in the same way as in (B).