Impaired systemic nucleocapsid antigen clearance in severe COVID‐19

Bauer, Christian; Mack, Elisabeth; Hefter, Véronique; Fischer, Alexandra; Volland, Kirsten; Skevaki, Chrysanthi; Neubauer, Andreas; Gress, Thomas M.; Becker, Stephan; Keller, Christian

doi:10.1002/jmv.29032

Cited by 1 publication

(2 citation statements)

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“…(21) Commensurate with these accessory roles, impaired clearance of N is associated with worse patient outcomes. (22, 23)…”

Section: Introductionmentioning

confidence: 99%

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A narrow ratio of nucleic acid to SARS-CoV-2 N-protein enables phase separation

Laughlin,

Young,

Gonzalez-Gutierrez

et al. 2024

Preprint

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SARS-CoV-2 Nucleocapsid protein (N) is a viral structural protein that packages the 30kb genomic RNA inside virions and forms condensates within infected cells through liquid-liquid phase separation (LLPS). N, in both soluble and condensed forms, has accessory roles in the viral life cycle including genome replication and immunosuppression. The ability to perform these tasks depends on phase separation and its reversibility. The conditions that stabilize and destabilize N condensates and the role of N-N interactions are poorly understood. We have investigated LLPS formation and dissolution in a minimalist system comprised of N protein and an ssDNA oligomer just long enough to support assembly. The short oligo allows us to focus on the role of N-N interaction. We have developed a sensitive FRET assay to interrogate LLPS assembly reactions from the perspective of the oligonucleotide. We find that N alone can form oligomers but that oligonucleotide enables their assembly into a three-dimensional phase. At a ~1:1 ratio of N to oligonucleotide LLPS formation is maximal. We find that a modest excess of N or of nucleic acid causes the LLPS to break down catastrophically. Under the conditions examined here assembly has a critical concentration of about 1 micromolar. The responsiveness of N condensates to their environment may have biological consequences. A better understanding of how nucleic acid modulates N-N association will shed light on condensate activity and could inform antiviral strategies targeting LLPS.

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“…(21) Commensurate with these accessory roles, impaired clearance of N is associated with worse patient outcomes. (22, 23)…”

Section: Introductionmentioning

confidence: 99%

“…(21) Commensurate with these accessory roles, impaired clearance of N is associated with worse patient outcomes. (22,23) N is comprised of two ordered domains flanked by three intrinsically disordered domains (IDR's; Figure 1A). The C-terminal ordered domain (CTD) forms a dimerization interface and also binds nucleic acid.…”

Section: Introductionmentioning

confidence: 99%