SARS-CoV-2 nucleocapsid protein interacts with immunoregulators and stress granules and phase separates to form liquid droplets.
Syam Prakash SomasekharanMartin GleavePublished in: FEBS letters (2021)
The current work investigated SARS-CoV-2 Nucleocapsid (NCAP or N protein) interactors in A549 human lung cancer cells using a SILAC-based mass spectrometry approach. NCAP interactors included proteins of the stress granule (SG) machinery and immunoregulators. NCAP showed specific interaction with the SG proteins G3BP1, G3BP2, YTHDF3, USP10 and PKR, and translocated to SGs following oxidative stress and heat shock. Treatment of recombinant NCAP with RNA isolated from A549 cells exposed to oxidative stress-stimulated NCAP to undergo liquid-liquid phase separation (LLPS). RNA degradation using RNase A treatment completely blocked the LLPS property of NCAP as well as its SG association. The RNA intercalator mitoxantrone also disrupted NCAP assembly in vitro and in cells. This study provides insight into the biological processes and biophysical properties of the SARS-CoV-2 NCAP.
Keyphrases
- sars cov
- induced apoptosis
- respiratory syndrome coronavirus
- oxidative stress
- heat shock
- mass spectrometry
- endoplasmic reticulum stress
- cell cycle arrest
- endothelial cells
- dna damage
- liquid chromatography
- binding protein
- high resolution
- combination therapy
- ionic liquid
- amino acid
- stress induced
- cell free
- cell proliferation
- atomic force microscopy
- capillary electrophoresis