Fig. 7: eIF4A1/2 destabilizes RNA condensates. | Nature Communications

Fig. 7: eIF4A1/2 destabilizes RNA condensates.

From: The DEAD-box helicase eIF4A1/2 acts as RNA chaperone during mitotic exit enabling chromatin decondensation

Fig. 7

A In vitro phase separation of 10 µM recombinant EGFP-DDX18, EGFP-DDX21, or EGFP-DDX27 in the presence or absence of 1 mg/ml RNA. Confocal images are representative of three independent experiments. B In vitro phase separation of 10 µM recombinant EGFP-DDX18, EGFP-DDX21, or EGFP-DDX27 (green in the presence of 1 mg/ml RNA and 0.5 µM mCherry-eIF4A1. Confocal images are representative of three independent experiments. Single RNA condensates are marked and shown as a close-up in the EGFP (green), mCherry (red), and merged channel (yellow as an overlay of red and green). On the right, line scans through a condensate is shown (EGFP signal in green, mCherry signal in red). Scale bars: 10 µm. C In vitro phase separation of 10 µM recombinant EGFP-DDX18 or EGFP-DDX21 and mCherry-DDX27 in the presence of 1 mg/ml RNA. Confocal images are representative of three independent experiments. Single RNA condensates are marked and shown as a close-up in the EGFP (green), mCherry (red), and merged channel (yellow as an overlay of red and green). On the right line, scans through a condensate is shown (EGFP signal in green, mCherry signal in red). D In vitro phase separation in the presence of RNA of the experiments as in (A) with increasing concentration of recombinant eIF4A1. Confocal images are representative of three independent experiments. E In vitro phase separation of 10 µM recombinant EGFP-DDX18 in the presence of 1 mg/ml RNA with increasing concentration of recombinant eIF4A1, eIF4A2, or eIF4A3. Confocal images are representative of three independent experiments. F In vitro phase separation of 10 µM recombinant EGFP-DDX18 in the presence of 1 mg/ml RNA with increasing concentration of the recombinant ATPase deficient E183Q or RNA-binding deficient R362/365Q mutant of eIF4A1. Confocal images are representative of three independent experiments. G Working model of eIF4A1/2 function as RNA mobilizers: As cytosolic RNA chaperone, eIF4A1 /2 facilitates the formation and/or maintenance of the perichromatin layer by enhancing RNA dynamics, thereby promoting their integration into the perichromatin layer. Scale bars: 10 µm. Source data are provided as a Source Data file.

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