Fig. 5: Intermolecular interactions between RNA and protein components tune the morphology of coexisting condensates. | Nature Communications

Fig. 5: Intermolecular interactions between RNA and protein components tune the morphology of coexisting condensates.

From: Sequence-encoded and composition-dependent protein-RNA interactions control multiphasic condensate morphologies

Fig. 5

a State diagram of PLP-KRP mixtures as a function of KRP-to-PLP ratio (mole: mole), showing that KRP, [KGKGG]5, does not affect PLP phase-separation (compare with Fig. 1a for RRP-PLP mixtures). b PLP partition coefficient (n = 60 droplets per sample) and diffusion rate (n = 3 droplets per sample) in PLP-KRP condensates as a function of KRP-to-PLP mixing ratio (mole: mole). The error bars represent the range of data (1–99%) for the bottom panel while the red line represents the mean value. Source data are provided as a Source Data file. c Fluorescence images showing that the morphology of coexisting PLP homotypic condensates and KRP-RNA condensates is non-engulfing and does not vary with RNA-to-KRP stoichiometry. Each type of droplet was separately prepared at initial concentrations of [FUSPLP] = 400 µM, [KGKGG]5 = 4 mg/ml, and variable poly(rU) RNA-to-KRP ratios (wt/wt), as indicated, and mixed (1:1 vol/vol). d A schematic diagram showing that due to insignificant KRP-PLP interfacial interactions, the PLP homotypic and KRP-RNA heterotypic condensates do not share any interface (non-engulfment) at both low and high RNA. e Domain architecture of FUSFL showing both PLP and RBD modules. f Fluorescence microscopy images and intensity profiles for coexisting homotypic FUSFL droplets (red) and heterotypic RRP-RNA condensates at different RNA-to-RRP ratio. Each type of droplet was separately prepared at initial concentrations of [FUSFL] = 21.3 µM, [FUSRGG3] = 1 mg/ml and variable poly(rU) RNA-to-RRP ratios (wt/wt), as indicated, and mixed (1:1 vol/vol). g Fluorescence micrographs and intensity profiles for Janus droplets formed by coexisting homotypic FUSFL droplets (red) and heterotypic KRP-RNA condensates (green). Each type of droplet was separately prepared at initial concentrations of [FUSFL] = 22 µM, [KGKGG5] = 4 mg/ml and poly(rU) = 3 mg/ml and mixed (1:1 vol/vol). All samples were made in a buffer containing 25 mM Tris-HCl, 150 mM NaCl, and 20 mM DTT. Scale bars represent 10 µm for (c, g) and 2 µm for (f).

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