Characterization of Gene Expression Suppression by Bovine Coronavirus Non-Structural Protein 1

Ohkami, Takehiro; Kitashin, Ichika; Kawashima, Riko; Yoshida, Aimi; Saito, Taizo; Takashima, Yasuhiro; Kamitani, Wataru; Nakagawa, Keisuke

doi:10.3390/v17070978

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Open AccessArticle

Characterization of Gene Expression Suppression by Bovine Coronavirus Non-Structural Protein 1

by

Takehiro Ohkami

^1,†,

Ichika Kitashin

¹,

Riko Kawashima

¹,

Aimi Yoshida

²,

Taizo Saito

^3,4,

Yasuhiro Takashima

^2,4,5,6,

Wataru Kamitani

⁷ and

Keisuke Nakagawa

^{1,2,4,6,*,†}

¹

Laboratory of Veterinary Microbiology, Joint Department of Veterinary Medicine, Gifu University, Yanagido, Gifu 501-1193, Japan

²

Joint Graduate School of Veterinary Sciences, Gifu University, Yanagido, Gifu 501-1193, Japan

³

Laboratory of Infectious Diseases, Joint Department of Veterinary Medicine, Gifu University, Yanagido, Gifu 501-1193, Japan

⁴

Education and Research Center for Food Animal Health, Gifu University, Yanagido, Gifu 501-1193, Japan

⁵

Laboratory of Veterinary Parasitology, Joint Department of Veterinary Medicine, Gifu University, Yanagido, Gifu 501-1193, Japan

⁶

Center for One Medicine Innovative Translational Research, Gifu University, Yanagido, Gifu 501-1193, Japan

⁷

Department of Infectious Diseases and Host Defense, Graduate School of Medicine, Gunma University, Gunma 371-8511, Japan

^*

Author to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Viruses 2025, 17(7), 978; https://doi.org/10.3390/v17070978 (registering DOI)

Submission received: 11 May 2025 / Revised: 11 July 2025 / Accepted: 12 July 2025 / Published: 13 July 2025

(This article belongs to the Special Issue Viral Strategies and Cellular Countermeasures That Regulate mRNA Access to the Translation Apparatus: 2nd Edition)

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Abstract

Coronavirus non-structural protein 1 (nsp1) is a pathogenic determinant of Betacoronaviruses. Previous studies demonstrated that the nsp1 of various coronaviruses induces host shutoff through a variety of mechanisms; however, there is little information on the function of bovine coronavirus (BCoV) nsp1. We aimed to characterize the host gene expression suppression function of BCoV nsp1. We first confirmed that the expression of BCoV nsp1 in MAC-T cells, a bovine mammary epithelial cell line, suppressed host and reporter gene expression. Subsequently, lysine and phenylalanine at amino acid positions 232 and 233, respectively, were identified as key residues required for this suppressive effect. Expression levels of housekeeping genes are comparable in cells expressing wild-type BCoV nsp1 and a mutant with alanine substitutions at positions 232 and 233 (BCoV nsp1-KF). Wild-type BCoV nsp1 localized to both the cytoplasm and nucleus; however, BCoV nsp1-KF exhibited prominent nuclear accumulation with dot-like structures. Using confocal microscopy and co-sedimentation analysis, we identified an association between wild-type BCoV nsp1, but not BCoV nsp1-KF, and ribosomes, suggesting that ribosome binding is required for BCoV nsp1-mediated suppression of host gene expression. This is the first study of the characterization of host gene expression suppression by BCoV nsp1.

Keywords: bovine coronavirus; non-structural protein 1; shutoff function; ribosome,

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MDPI and ACS Style

Ohkami, T.; Kitashin, I.; Kawashima, R.; Yoshida, A.; Saito, T.; Takashima, Y.; Kamitani, W.; Nakagawa, K. Characterization of Gene Expression Suppression by Bovine Coronavirus Non-Structural Protein 1. Viruses 2025, 17, 978. https://doi.org/10.3390/v17070978

AMA Style

Ohkami T, Kitashin I, Kawashima R, Yoshida A, Saito T, Takashima Y, Kamitani W, Nakagawa K. Characterization of Gene Expression Suppression by Bovine Coronavirus Non-Structural Protein 1. Viruses. 2025; 17(7):978. https://doi.org/10.3390/v17070978

Chicago/Turabian Style

Ohkami, Takehiro, Ichika Kitashin, Riko Kawashima, Aimi Yoshida, Taizo Saito, Yasuhiro Takashima, Wataru Kamitani, and Keisuke Nakagawa. 2025. "Characterization of Gene Expression Suppression by Bovine Coronavirus Non-Structural Protein 1" Viruses 17, no. 7: 978. https://doi.org/10.3390/v17070978

APA Style

Ohkami, T., Kitashin, I., Kawashima, R., Yoshida, A., Saito, T., Takashima, Y., Kamitani, W., & Nakagawa, K. (2025). Characterization of Gene Expression Suppression by Bovine Coronavirus Non-Structural Protein 1. Viruses, 17(7), 978. https://doi.org/10.3390/v17070978

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

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Characterization of Gene Expression Suppression by Bovine Coronavirus Non-Structural Protein 1

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