Evidence that AGAUA and CCAGA initiate translation in the same mRNA in region E3 of adenovirus

Gene delivery from the E3 region of replicating human adenovirus: evaluation of the 6.7 K/gp19 K region

“…While both the 6.7 K and gp19 K genes are encoded by the same mRNA, the initial timing of transgene expression would be predicted to most closely mimic that of the gp19 K gene. 16 In the wild-type Ad5 infection, synthesis of gp19 K was detected 8 h p.i. by Western blot (Figure 2a).…”

“…Primary antibodies reacting against H-ras (mouse monoclonal F235; Santa Cruz (Santa Cruz, CA, USA) used at 0.1 mg/ml), p16 (rabbit polyclonal C-20; Santa Cruz used at 0.1 mg/ml), mTNF (goat polyclonal; R&D systems (Minneapolis, MN, USA) used at 0.1 g/ml), E3-gp19 K, 16 E3-ADP (Hawkins and Wold, unpublished data), E3-RID␤, 52 E3-14.7 K 53 (all E3 antibodies are rabbit polyclonals obtained from Dr WSM Wold, used at a 1:500 dilution), or against the Ad5 virion (American Qualex, Los Angeles, CA, USA; used at 1:1000) were incubated for 1-2 h in blocking solution TNT (50 mm Tris, pH 8.0, 150 mm NaCl, and 0.1% Tween-20) plus 5% non-fat dry milk. The membranes were incubated with secondary anti-mouse or anti-rabbit IgG antibodies conjugated to horseradish peroxidase (Amersham, used at 1:1000 dilution) for 1 h at 25°C and the signal was visualized using either the ECL or ECL plus system (Amersham).…”

“…These two genes were taken out together because they are adjacent to one another in the viral genome, are encoded by the same E3 mRNA, and have overlapping open reading frames. 16 By deleting both genes, the capacity of the viral genome to package a foreign gene is extended. The E3 6.7 K protein functions in concert with the RID complex to reduce TRAIL receptor-1 and -2 cell surface expression, 17 the gp19 K protein is known to bind MHC class I molecules in the endoplasmic reticulum (ER), inhibiting their transport to the infected cell surface (reviewed in Ref.…”

Three Adenovirus E3 Proteins Cooperate to Evade Apoptosis by Tumor Necrosis Factor-related Apoptosis-inducing Ligand Receptor-1 and -2

“…All viruses used in these experiments have been described in detail previously (27,28) and were kind gifts of W. Wold (St. Louis University, St. Louis, MO). Briefly, rec700 is a recombinant Ad5 subtype virus containing the Ad2 10.4K protein (EcoRI-D fragment, map position 73-86) and the Ad5 14.5K and 14.7K proteins.…”

Down-regulation of human adenovirus E1a by E3 gene products: evidence for translational control of E1a by E3 14{middle dot}5K and/or E3 10{middle dot}4K products

“…Over many experiments the relative levels of E l a proteins vary considerably but range from about fivefold to greater than 12-fold more E l a in mutant compared to wt Ad 5-infected cells. Similar results were obtained when E l a levels in wt Ad 2-infected cells were compared with the Ad 2 E3 deletion mutant dl 801 (Ad 2 is another member of the group C adenoviruses), but not when compared with the hybrid virus rec 700 (Zhang et at., 1992) in which the E3 region from Ad 2 is cloned into an Ad 5 genome lacking its own E3 region (Wold et al, 1986). The reason for this apparent serotype specificity is unclear at this time).…”