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. 2017 Nov 30;6:e31476. doi: 10.7554/eLife.31476

Figure 1. ATP hydrolysis by eIF4A stimulates recruitment of natural mRNA RPL41A as well as a synthetic 50-mer made up largely of CAA-repeats, presumed to be unstructured, with an AUG start codon 23 nucleotides from the 5'-end (CAA 50-mer).

The concentration of ATP and analogs was 2 mM. (A) Percentage of RPL41A recruited to the PIC versus time. (B) Percentage of CAA 50-mer recruited to the PIC versus time. The larger plot shows the time-course up to 20 min. The inset shows the first 1.5 min for clarity. With the CAA 50-mer in the presence of ATP and eIF4A, reaction rates are at the limit of our ability to measure by hand but the results provide an estimate of the value. (C) Observed rate constants (kobs) and reaction endpoints from the data in panels A and B. All data in the figure are mean values (n ≥ 2) and error bars represent average deviation of the mean. With RPL41A, rates with ADPNP, ADPCP, ADP, no nucleotide, and no eIF4A could not be measured accurately due to low endpoints (‘ND’).

Figure 1—source data 1. Individual measurements of the percent of RPL41A and CAA 50-mer recruited to the PIC over time.
elife-31476-fig1-data1.xlsx (43.7KB, xlsx)
DOI: 10.7554/eLife.31476.004

Figure 1.

Figure 1—figure supplement 1. RPL41A can form structural elements throughout its length whereas the CAA 50-mer has a low propensity to form structures.

Figure 1—figure supplement 1.

RNA folds, predicted by mfold (Zuker, 2003), that are representative of the population of conformations adopted by (A–B) CAA 50-mer, ∆G > −1 kcal/mol; (C) RPL41A, ∆G = −46 kcal/mol; (D) chimeric RNA with CAA-repeat 5’-UTR and RPL41A sequence 3’ of the AUG codon (RNA 7; Figure 3A), ∆G = −51 kcal/mol.