in cell phase separation assay -NP
(Iserman 2020 BioRxiv PMID: 32587965)
N-protein phase separates in mammalian cells and can be disrupted by small molecules
co-transfected HEK293 cells with N-protein fused to GFPspark and with H2B:mCherry
Cells with higher levels of transfection were more likely to form spherical droplets in the cytoplasm - N-protein condensation is concentration dependent
N-protein droplets readily underwent fusion and recovered quickly following FRAP - liquid-like condensates
cellular localisation
N-protein signal was generally excluded from the nucleus
disruption of condensates in cells by the known candidates
1,6-hexanediol
The aminoglycoside kanamycin binds promiscuously to nucleic acids via electrostatic interactions and, is an interesting candidate to disrupt LLPS due to its conformational adaptability and thus low RNA target specificity and was implicated as antiviral in HIV-1 by preventing RNA-protein interactions (21).