Natural mutations in key NLS amino acids regulate nucleoplasmic shuttling and replication efficiency in PRRSV
Highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) is a mutant strain of the classic porcine reproductive and respiratory syndrome virus (PRRSV) characterized by high morbidity and mortality rates. Epidemiological analysis revealed a natural mutation and stable inherita...
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Frontiers Media S.A.
2025-07-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1587634/full |
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| author | Xianchang Zhu Yang Xia Qian Lei Yu Gan Shenghai Jiang Lian Huang Qihu Wen Wei Fu Bo Zhang Yi Zhang Shanshan Xie Jida Li Jida Li |
| author_facet | Xianchang Zhu Yang Xia Qian Lei Yu Gan Shenghai Jiang Lian Huang Qihu Wen Wei Fu Bo Zhang Yi Zhang Shanshan Xie Jida Li Jida Li |
| author_sort | Xianchang Zhu |
| collection | DOAJ |
| description | Highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) is a mutant strain of the classic porcine reproductive and respiratory syndrome virus (PRRSV) characterized by high morbidity and mortality rates. Epidemiological analysis revealed a natural mutation and stable inheritance of amino acid 46 (41-PGKKNKK-47 mutated to 41-PGKKNRK-47) in the nuclear localization signal or sequence (NLS) region of the N protein of HP-PRRSV. In this study, we showed that the nucleoplasmic shuttling of the HP-PRRSV N protein was associated with a higher efficiency of viral replication than that of the classical PRRSV. The nuclear transporter receptors KPNB1, KPNA1, KPNA2, KPNA6, and KPNA7 were involved in the nuclear import of the N protein. Additionally, the mRNA expression levels of KPNB1 and KPNA1 differed between the two strains after infecting the Marc-145 cells with these strains. The viral replication efficiency also decreased when expression levels of KPNA1 and/or KPNB1 were lowered. Finally, protein binding simulation and kinetic assay showed that the mutation of key amino acid 46 in the NLS region altered the binding mode and kinetics of the N proteins to KPNA1 and KPNB1. This study elucidates, for the first time, the reasons for the enhanced nucleoplasmic shuttling and replication efficiency of HP-PRRSV from the perspective of protein entry into the nucleus. It also provides a foundational reference for the prevention and control of PRRSV. |
| format | Article |
| id | doaj-art-c3311fc5c7874447b7f0b4cd8c5652cd |
| institution | DOAJ |
| issn | 1664-302X |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Microbiology |
| spelling | doaj-art-c3311fc5c7874447b7f0b4cd8c5652cd2025-08-20T02:44:27ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-07-011610.3389/fmicb.2025.15876341587634Natural mutations in key NLS amino acids regulate nucleoplasmic shuttling and replication efficiency in PRRSVXianchang Zhu0Yang Xia1Qian Lei2Yu Gan3Shenghai Jiang4Lian Huang5Qihu Wen6Wei Fu7Bo Zhang8Yi Zhang9Shanshan Xie10Jida Li11Jida Li12Institute of Zoonosis, College of Public Health, Zunyi Medical University, Zunyi, Guizhou, ChinaSouthwest Guizhou Vocational and Technical College for Nationalities, Xingyi, Guizhou, ChinaInstitute of Zoonosis, College of Public Health, Zunyi Medical University, Zunyi, Guizhou, ChinaInstitute of Zoonosis, College of Public Health, Zunyi Medical University, Zunyi, Guizhou, ChinaDepartment of Preventive Health Care, Yunan District People's Hospital, Yunfu, Guangdong, ChinaInstitute of Zoonosis, College of Public Health, Zunyi Medical University, Zunyi, Guizhou, ChinaInstitute of Zoonosis, College of Public Health, Zunyi Medical University, Zunyi, Guizhou, ChinaInstitute of Zoonosis, College of Public Health, Zunyi Medical University, Zunyi, Guizhou, ChinaCollege of Basic Medicine, Zunyi Medical University, Zunyi, Guizhou, ChinaInstitute of Zoonosis, College of Public Health, Zunyi Medical University, Zunyi, Guizhou, ChinaCollege of Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan, ChinaInstitute of Zoonosis, College of Public Health, Zunyi Medical University, Zunyi, Guizhou, ChinaKey Laboratory of Maternal & Child Health and Exposure Science of Guizhou Higher Education Institutes, Zunyi, Guizhou, ChinaHighly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) is a mutant strain of the classic porcine reproductive and respiratory syndrome virus (PRRSV) characterized by high morbidity and mortality rates. Epidemiological analysis revealed a natural mutation and stable inheritance of amino acid 46 (41-PGKKNKK-47 mutated to 41-PGKKNRK-47) in the nuclear localization signal or sequence (NLS) region of the N protein of HP-PRRSV. In this study, we showed that the nucleoplasmic shuttling of the HP-PRRSV N protein was associated with a higher efficiency of viral replication than that of the classical PRRSV. The nuclear transporter receptors KPNB1, KPNA1, KPNA2, KPNA6, and KPNA7 were involved in the nuclear import of the N protein. Additionally, the mRNA expression levels of KPNB1 and KPNA1 differed between the two strains after infecting the Marc-145 cells with these strains. The viral replication efficiency also decreased when expression levels of KPNA1 and/or KPNB1 were lowered. Finally, protein binding simulation and kinetic assay showed that the mutation of key amino acid 46 in the NLS region altered the binding mode and kinetics of the N proteins to KPNA1 and KPNB1. This study elucidates, for the first time, the reasons for the enhanced nucleoplasmic shuttling and replication efficiency of HP-PRRSV from the perspective of protein entry into the nucleus. It also provides a foundational reference for the prevention and control of PRRSV.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1587634/fullPRRSVnucleocapsid proteinNLSNTRnuclear-mass shuttle |
| spellingShingle | Xianchang Zhu Yang Xia Qian Lei Yu Gan Shenghai Jiang Lian Huang Qihu Wen Wei Fu Bo Zhang Yi Zhang Shanshan Xie Jida Li Jida Li Natural mutations in key NLS amino acids regulate nucleoplasmic shuttling and replication efficiency in PRRSV Frontiers in Microbiology PRRSV nucleocapsid protein NLS NTR nuclear-mass shuttle |
| title | Natural mutations in key NLS amino acids regulate nucleoplasmic shuttling and replication efficiency in PRRSV |
| title_full | Natural mutations in key NLS amino acids regulate nucleoplasmic shuttling and replication efficiency in PRRSV |
| title_fullStr | Natural mutations in key NLS amino acids regulate nucleoplasmic shuttling and replication efficiency in PRRSV |
| title_full_unstemmed | Natural mutations in key NLS amino acids regulate nucleoplasmic shuttling and replication efficiency in PRRSV |
| title_short | Natural mutations in key NLS amino acids regulate nucleoplasmic shuttling and replication efficiency in PRRSV |
| title_sort | natural mutations in key nls amino acids regulate nucleoplasmic shuttling and replication efficiency in prrsv |
| topic | PRRSV nucleocapsid protein NLS NTR nuclear-mass shuttle |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1587634/full |
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