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Conserved nucleotides in the terminus of the 3′ UTR region are important for the replication and infectivity of porcine reproductive and respiratory syndrome virus

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Abstract

The 3′ untranslated region (3′ UTR), including the poly (A) tail, reportedly plays an important role in arterivirus replication, but the roles of the cis-acting elements present in the 3′ UTR of porcine reproductive and respiratory syndrome virus (PRRSV) remain largely unknown. In the present study, PCR-based mutagenic analysis was conducted on the 3′ UTR of PRRSV infectious full-length cDNA clone pAPRRS to investigate the structure and function of the conserved terminal nucleotides between the poly (A) tail and the 3′ UTR region. Our findings indicated that the conservation of the primary sequence of the 3′ terminal nucleotides, rather than the surrounding secondary structure, was vital for viral replication and infectivity. Four nucleotides (nt) (5′-15517AAUU15520-3′) at the 3′ proximal end of the 3′ UTR and the dinucleotide 5′-AU-3′ exerted an important regulatory effect on viral viability. Of the five 3′-terminal nucleotides of the 3′ UTR (5′-15503AACCA15507-3′), at least three, including the last dinucleotide (5′-CA-3′), were essential for maintaining viral infectivity. Taken together, the 3′-terminal conserved sequence plays a critical role in PRRSV replication and may function as a contact site for specific assembly of the replication complex.

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Acknowledgements

This study was partially supported by a grant from the Natural Sciences Foundation of China (30972204, 30901078 and 31100121), the Basic Research Project grant (2013JB02), the EU Frame 7 Program Project (245141), National High Tech Plan (863 Plan) (2011AA10A208-2), NSFC-Guangdong Joint Foundation (U0931003), International Sci & Tech Cooperation Program (2010DFB33920). We thank Dr. Ying Fang of South Dakota State University for generously providing the monoclonal antibody against the PRRSV N protein.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Swine Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No. 518, Ziyue Road, Minhang District, Shanghai, 200241, People’s Republic of China

    Yang Yin, Changlong Liu, Ping Liu, Zuzhang Wei, Jiaqi Lu, Guangzhi Tong, Fei Gao & Shishan Yuan

  2. Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen, Guangdong Province, 518045, People’s Republic of China

    Yang Yin

  3. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province, 210095, People’s Republic of China

    Yang Yin, Ping Liu & Huochun Yao

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  1. Yang Yin
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  2. Changlong Liu
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Correspondence to Guangzhi Tong or Fei Gao.

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Yin, Y., Liu, C., Liu, P. et al. Conserved nucleotides in the terminus of the 3′ UTR region are important for the replication and infectivity of porcine reproductive and respiratory syndrome virus. Arch Virol 158, 1719–1732 (2013). https://doi.org/10.1007/s00705-013-1661-3

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