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. Author manuscript; available in PMC: 2012 Mar 18.
Published in final edited form as: J Mol Biol. 2011 Jan 12;407(1):21–34. doi: 10.1016/j.jmb.2011.01.006

Figure 3.

Figure 3

Upstream initiation from GUG codon(s) in the HP FMR1 context. (A) Sequence and presumed structure of HP. GUG sequence is indicated in outlined font. (B) Autoradiograph of [35S] methionine-labeled, in vitro-translated FL protein products separated by SDS-PAGE. FMR15′UTR(HP)-FL initiates translation early and runs high, while there is no product from either 5″HP-FMR15′UTR(30 CGG)-FL or 5′HP-synthetic 5′UTR(30 CGG)-FL constructs. Abbreviation: Syn, Synthetic. (C) Autoradiograph of [35S]-labeled proteins produced from FMR15′UTR(HP)-FL mRNAs with and without GUG codons. U-to-A changes in the GUG sequences within the hairpin results in normal-length protein product. (D) In vitro translation of FMR1- and synthetic-5′UTR mRNAs with internal HPs. Results are given in RLUs of FL expression and are not normalized to RL levels. GUG: FMR15′UTR(HP)-FL; GAG: FMR15′UTR(HP/GAG)-FL; GUG (stop): synthetic 5′UTR(HP)-FL; GUG (no stop): synthetic 5′UTR(HP/GUG)[no stop]-FL. All messages were translated at least in triplicate.