Fig. 5: Upstream ATG, uORF, and Kozak sequences.
From: Redefining GBA gene structure unveils the ability of Cap-independent, IRES-dependent gene regulation
a Positions of uATGs, uORFs, and their Kozak sequences within the region from exon 1 to exon 5. The ATGs are numbered from the 5’ ends of the P1 promoter-controlled transcripts. Two reported start sites (#8 and #10) are indicated in red, and the columns are highlighted in yellow9,20. The numbers in the table are the predicted sizes of the ORFs for the indicated ATGs on the top line. b Schematic of three kinds of v3 constructs inserted into expression vectors. FLAG tags were added to all constructs at the 3’ end. Top line, CDS only; second line (C), CDS with the original 5’ UTR containing a weak Kozak sequence (V3); third line, CDS with a partial 5’ UTR containing an adequate Kozak sequence (A); fourth line, CDS with a partial 5’ UTR containing a strong Kozak sequence (S). c FLAG-tagged GCase expression with or without glycosylation in HEK293 cells. GCase protein expression was detected by western blot analysis with anti-FLAG antibodies. C, V3, A, and S indicate the same as in b. pCI, empty vector; H, HEK293 cells without transfection. The molecular sizes are indicated on the right side. The experiment was performed in triplicate and the representative data was shown.
