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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 7. DDX3X missense mutations alter translation of select targets.

Figure 7.

A, Western blots depicting DDX3X levels (WT, R326H, or R376C) with simultaneous knockdown of endogenous DDX3X. Endogenous and FLAG-tagged DDX3X is detected. B, C, Translation of in vitro transcribed reporter RNAs in lysates expressing either DDX3X WT, R326H (B) or R376C (C) Signal was normalized to WT. Predicted DDX3X sensitive reporters (ATF5, CCNE1, RPLP1, PRKRA and RAC1) show robust decreases in translation, while control transcripts (SIKE1, SRSF5) right of dotted line demonstrate either no change or modest increases. Box and whisker plot with bars indicating data range, box at upper and lower quartiles, and line at the median. D, Proposed model for mechanism of DDX3X mutants, based upon in vitro biochemical and cell biology studies. Mild missense or DDX3X LoF are impair translation of some targets but do not induce granule formation. Severe DDX3X missense mutations show impaired RNA release and helicase activity, and altered translation of DDX3X-dependent targets. This results in sequestration of RNAs and RNA binding proteins and formation of aberrant RNP granules. Created with BioRender.com.