Human Post-Translational SUMOylation Modification of SARS-CoV-2 Nucleocapsid Protein Enhances Its Interaction Affinity with Itself and Plays a Critical Role in Its Nuclear Translocation

Abstract

Viruses, such as Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), infect hosts and take advantage of host cellular machinery for genome replication and new virion production. Identifying and elucidating host pathways for viral infection is critical for understanding the development of the viral life cycle and novel therapeutics. The SARS-CoV-2 N protein is critical for viral RNA (vRNA) genome packaging in new virion formation. Using our quantitative Förster energy transfer/Mass spectrometry (qFRET/MS) coupled method and immunofluorescence imaging, we identified three SUMOylation sites of the SARS-CoV-2 N protein. We found that (1) Small Ubiquitin-like modifier (SUMO) modification in Nucleocapsid (N) protein interaction affinity increased, leading to enhanced oligomerization of the N protein; (2) one of the identified SUMOylation sites, K65, is critical for its nuclear translocation. These results suggest that the host human SUMOylation pathway may be critical for N protein functions in viral replication and pathology in vivo. Thus, blocking essential host pathways could provide a novel strategy for future anti-viral therapeutics development, such as for SARS-CoV-2 and other viruses.

A viral assembly inhibitor blocks SARS-CoV-2 replication in airway epithelial cells

“…SARS-CoV-2 infection increased the expression of the PARP9, PARP14, and PARP10 genes, but SARS-CoV-2 infection in the presence of PAV-104 treatment down-regulated the expression of these genes (as compared to untreated/uninfected control). Beyond host factors defined by Reactome as members of the maturation of nucleoprotein pathway, PIAS1, and SRPK3, which have been reported to play roles in SUMOylating 42 and phosphorylating SARS-CoV-2 N 43 , respectively, were significantly down-regulated in the SARS-CoV-2 infection with PAV-104 treatment group (Supplementary Fig. 2c ).…”