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. 2001 Nov 1;29(21):4341–4351. doi: 10.1093/nar/29.21.4341

Figure 6.

Figure 6

Translational repression by the Chop uORF is not dependent on the intercistronic region. Expression constructs containing the 5′UTR either in a wild-type form (5) or mutated on the uAUGs (6) were used to mutate the intercistronic region. The arrows represent point mutation of the uAUGs. When present the uORF is boxed. These constructs were transfected in HeLa cells then, 48 h after transfection, cells were harvested. The relative LUC activities and mRNA levels were analyzed as described above. In plasmid ICmt, the CHOP intercistronic region was replaced by a LacZ sequence which does not contain any AUG [(3) and (4)]. The initiation context of the LUC AUG was not changed. Plasmids mutated on the AUG#3 are given as a control (1) (2). A spacer of 47 bp [(7) and (8)] or 140 bp [(9) and (10)] that does not contain AUG codon was introduced into the intercistronic region of the pCMV–5′UTR–LUC plasmid mutated or not on the uAUGs. In the constructs ΔIC, the intercistronic region was deleted [(11) and (12)]. The mutations do not affect the Kozak consensus sequence of the LUC AUG.