SARS-CoV-2 nsp15 enhances viral virulence by subverting host antiviral defenses

A Allen Caobi (Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine) C Chia-Ming Su (Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine) C Christian M. Beusch D Devin Kenney (Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine) T Tamarand L. Darling (Department of Medicine, Washington University School of Medicine) S Shuchen Feng (Department of Microbiology and Immunology, Loyola University Chicago Stritch School of Medicine) M Marc Semaan (Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine) A Alan Wacquiez (Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine) N Nathan L. Sanders (The Pulmonary Center, Boston University Chobanian and Avedisian School of Medicine) E Ena S. Tully (Department of Biochemistry, Institute for Molecular Virology, Center for Quantitative Cell Imaging, University of Wisconsin-Madison) D Da-Yuan Chen (Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine) M Monika Evdokimova (Department of Microbiology and Immunology, Loyola University Chicago Stritch School of Medicine) Z Zhen Ding (Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine) D Dakota Jones K Konstantinos-Dionysios Alysandratos J Joseph P. Mizgerd (National Emerging Infectious Diseases Laboratories, Boston University) R Robert N. Kirchdoerfer D Darrell N. Kotton F Florian Douam (National Emerging Infectious Diseases Laboratories, Boston University) N Nicholas A. Crossland (National Emerging Infectious Diseases Laboratories, Boston University) A Adrianus C. M. Boon (Department of Medicine, Washington University School of Medicine) D David E. Gordon S Susan C. Baker (Department of Microbiology and Immunology, Loyola University Chicago Stritch School of Medicine) M Mohsan Saeed (Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine)

Abstract

SARS-CoV-2 encodes numerous virulence factors, yet their precise mechanisms of action remain unknown. We provide evidence that the SARS-CoV-2 nonstructural protein 15 (nsp15) enhances viral virulence by suppressing the production of viral double-stranded (dsRNA), a potent inducer of antiviral signaling. The viral variants lacking nsp15 endoribonuclease activity elicited higher innate immune responses and exhibited reduced replication in human stem cell–derived lung alveolar type II epithelial cells, as well as in the lungs of infected hamsters. Consistently, these variants caused significantly less weight loss and mortality compared to wild-type (WT) virus in K18-hACE2 mice. Mechanistically, the cells infected with nsp15 mutants accumulated more viral dsRNA, causing enhanced stimulation of the interferon pathway. Chemical inhibition of interferon signaling dampened immune responses to nsp15 mutants and restored their replication to levels similar to the WT virus. These findings indicate that the endoribonuclease activity of nsp15 contributes to viral virulence by limiting the accumulation of viral dsRNA, thereby allowing robust replication with reduced activation of the host innate immune response.

Article Details

Volume / Issue Vol. 122, Issue 24
Published June 17, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (24)

A

Allen Caobi

Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine

C

Chia-Ming Su

Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine

C

Christian M. Beusch

D

Devin Kenney

Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine

T

Tamarand L. Darling

Department of Medicine, Washington University School of Medicine

S

Shuchen Feng

Department of Microbiology and Immunology, Loyola University Chicago Stritch School of Medicine

M

Marc Semaan

Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine

A

Alan Wacquiez

Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine

N

Nathan L. Sanders

The Pulmonary Center, Boston University Chobanian and Avedisian School of Medicine

E

Ena S. Tully

Department of Biochemistry, Institute for Molecular Virology, Center for Quantitative Cell Imaging, University of Wisconsin-Madison

D

Da-Yuan Chen

Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine

M

Monika Evdokimova

Department of Microbiology and Immunology, Loyola University Chicago Stritch School of Medicine

Z

Zhen Ding

Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine

D

Dakota Jones

K

Konstantinos-Dionysios Alysandratos

J

Joseph P. Mizgerd

National Emerging Infectious Diseases Laboratories, Boston University

R

Robert N. Kirchdoerfer

D

Darrell N. Kotton

F

Florian Douam

National Emerging Infectious Diseases Laboratories, Boston University

N

Nicholas A. Crossland

National Emerging Infectious Diseases Laboratories, Boston University

A

Adrianus C. M. Boon

Department of Medicine, Washington University School of Medicine

D

David E. Gordon

S

Susan C. Baker

Department of Microbiology and Immunology, Loyola University Chicago Stritch School of Medicine

M

Mohsan Saeed

Department of Biochemistry and Cell Biology, Boston University Chobanian and Avedisian School of Medicine