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J Struct Biol . Characterization of SARS-CoV-2 nucleocapsid protein oligomers

tetano

Editor, Senior Moderator
J Struct Biol


. 2024 Dec 13:108162.
doi: 10.1016/j.jsb.2024.108162. Online ahead of print. Characterization of SARS-CoV-2 nucleocapsid protein oligomers

Domenica Farci[SUP] 1 [/SUP], André T Graça[SUP] 2 [/SUP], Michael Hall[SUP] 2 [/SUP], Patrycja Haniewicz[SUP] 3 [/SUP], Sami Kereïche[SUP] 4 [/SUP], Peter Faull[SUP] 5 [/SUP], Joanna Kirkpatrick[SUP] 6 [/SUP], Enzo Tramontano[SUP] 7 [/SUP], Wolfgang P Schröder[SUP] 2 [/SUP], Dario Piano[SUP] 8 [/SUP]



Affiliations
Abstract

Oligomers of the SARS-CoV-2 nucleocapsid (N) protein are characterized by pronounced instability resulting in fast degradation. This property likely relates to two contrasting behaviors of the N protein: genome stabilization through a compact nucleocapsid during cell evasion and genome release by nucleocapsid disassembling during infection. In vivo, the N protein forms rounded complexes of high molecular mass from its interaction with the viral genome. To study the N protein and understand its instability, we analyzed degradation profiles under different conditions by size-exclusion chromatography and characterized samples by mass spectrometry and cryo-electron microscopy. We identified self-cleavage properties of the N protein based on specific Proprotein convertases activities, with Cl[SUP]-[/SUP] playing a key role in modulating stability and degradation. These findings allowed isolation of a stable oligomeric complex of N, for which we report the 3D structure at ∼6.8 Å resolution. Findings are discussed considering available knowledge about the coronaviruses' infection cycle.

Keywords: Covid-19; Cryo-electron microscopy; Furin; Nucleocapsid protein; Proprotein convertases; SARS-CoV-2; Self-cleavage.

 
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