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

Vipul Madahar; Runrui Dang; Quanqing Zhang; Chuchu Liu; Victor G J Rodgers; Jiayu Liao

doi:10.3390/v15071600

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

Viruses. 2023 Jul 21;15(7):1600. doi: 10.3390/v15071600.

Authors

Vipul Madahar¹, Runrui Dang¹, Quanqing Zhang^{2

3}, Chuchu Liu¹, Victor G J Rodgers^{1

4}, Jiayu Liao^{1

2

4}

Affiliations

¹ Department of Bioengineering, College of Engineering, Bourns College of Engineering, University of California at Riverside, Riverside, CA 92521, USA.
² Institute for Integrative Genome Biology, University of California at Riverside, Riverside, CA 92521, USA.
³ Department of Botany, College of Natural & Agricultural Sciences, University of California at Riverside, Riverside, CA 92521, USA.
⁴ Biomedical Science, School of Medicine, University of California at Riverside, Riverside, CA 92521, USA.

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.

Keywords: SARS-CoV-2 N protein; SUMOylation; nuclear translocation; protein interaction dissociation constant KD; qFRET.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

COVID-19*
Humans
Protein Processing, Post-Translational
SARS-CoV-2
Sumoylation*
Virus Replication

Grants and funding

This work was partially funded by the UCR Academic Senate Research Grant and Attaisina Gift grant to J.L.