Fig. 2: Phase separation of SARS-CoV-2 nsp8. | Communications Biology

Fig. 2: Phase separation of SARS-CoV-2 nsp8.

From: Multiscale characterization reveals oligomerization dependent phase separation of primer-independent RNA polymerase nsp8 from SARS-CoV-2

Fig. 2

a LLPS assay of nsp8 dimer at 1 mg/mL in the buffer of 20 mM HEPES pH 7.4, 50 or 100 mM NaCl. b LLPS assay of nsp8 dimer at 2 mg/mL in the buffer of 20 mM HEPES pH 7.4, 100 or 150 mM NaCl. c Liquid-like droplets formed by RED-tris-NTA labeled nsp8 dimers at 1 mg/mL in the buffer of 20 mM HEPES pH 7.4, 50 mM NaCl, can be dissolved upon increasing NaCl concentration to 275 mM by mixing nsp8 dimer (1 mg/mL) dissolved in 500 mM NaCl with volume ratio 1:1. d In vitro FRAP analysis of the condensates formed by RED-tris-NTA labeled nsp8 at 1 mg/mL in a buffer of 20 mM HEPES pH 7.4, 50 mM NaCl. Data were representative of three independent experiments; error bars represent standard deviation. Source data is available in Supplementary Data 1. e nsp8 tetramer at 0.25 mg/mL forms solid-like condensates in the buffer of 20 mM HEPES pH 7.4, 50 mM NaCl. The solid-like condensates can be dissolved upon increasing NaCl concentration. f Deletion of N-terminal 76 residues inhibits the phase separation of nsp8 dimer in a buffer of 20 mM HEPES pH 7.4, 100 mM NaCl. The concentration of wildtype or mutated nsp8 dimer was 2 mg/mL. g Deletion of N-terminal 76 residues reduces the aggregation of nsp8 tetramer in a buffer of 20 mM HEPES pH 7.4, 50 mM NaCl. The concentration of wildtype or mutated nsp8 tetramer was 0.25 mg/mL.

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