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. Author manuscript; available in PMC: 2021 May 6.
Published in final edited form as: Neuron. 2020 Mar 4;106(3):404–420.e8. doi: 10.1016/j.neuron.2020.01.042

Figure 5. DDX3X missense mutants exhibit disrupted helicase activity.

Figure 5.

A, Left, Diagram of DDX3X activity tested in this assay. ATP hydrolysis is necessary for initial binding and release of RNA, but not for RNA unwinding. Right, Non-denaturing gel depicting time course of helicase assay in which amounts of dsRNA (not unwound by DDX3X) and ssRNA (unwinding by DDX3X) are measured. B, Unwinding assay for WT, A233V, T323I, R326H, R376C, I415del, R475G, I514T, and T532 DDX3X. C, D, Graphs depicting Vmax (C) and Km (D). Note, I415del and T532M had unwinding curves which did not vary within the range of tested RNA concentrations (0–40 uM) so Km was not determined (n.d.). The majority of mutants had lower Km (indicating higher affinity for RNA) than WT, with the exception of I514T. Error bars=SD.