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J. Vet. Sci. 2017; 18(S1): 307-313  
Effect of the fourth nucleotide at the 3' end of neuraminidase and matrix viral genomic RNA on the pathogenicity of influenza virus A/PR/8/34
Chung-Young Lee1,†, Hyuk-Joon Kwon2,3,4,†, Thanh Trung Nguyen1, Ilhwan Kim5, Hyung-Kwan Jang6, Jae-Hong Kim1,3,*
1Laboratory of Avian Diseases, 2Laboratory of Poultry Production Medicine, and 3Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul 08826, Korea
4Farm Animal Clinical Training and Research Center, Institutes of Green Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea
5Division of Antimicrobial Resistance, Center for Infectious Diseases, National Research Institute of Health, Korea Centers for Disease Control & Prevention (KCDC), Cheongju 28159, Korea
6Department of Infectious Diseases & Avian Diseases, College of Veterinary Medicine, Chonbuk National University, Iksan 54596, Korea
Correspondence to: Tel: +82-2-880-1250; Fax: +82-2-885-6614; E-mail: kimhong@snu.ac.kr
Received: May 8, 2017; Revised: July 10, 2017; Accepted: July 26, 2017; Published online: August 31, 2017.
Twelve nucleotides located at the 3′ end of viral genomic RNA (vRNA) are conserved among influenza A viruses (IAV) and have a promoter function. Hoffmann’s 8-plasmid reverse genetics vector system introduced mutations at position 4, C nucleotide (C4) to U nucleotide (U4), of the 3′ ends of neuraminidase (NA) and matrix (M) vRNAs of wild-type A/PR/8/34 (PR8). This resulted in a constellation of C4 and U4 vRNAs coding for low (polymerases) and relatively high (all others) copy number proteins, respectively. U4 has been reported to increase promoter activity in comparison to C4, but the constellation effect on the replication efficiency and pathogenicity of reverse genetics PR8 (rgPR8) has not been fully elucidated. In the present study, we generated 3 recombinant viruses with C4 in the NA and/or M vRNAs and rgPR8 by using reverse genetics and compared their pathobiological traits. The mutant viruses showed lower replication efficiency than rgPR8 due to the low transcription levels of NA and/or M genes. Furthermore, C4 in the NA and/or M vRNAs induced lower PR8 virus pathogenicity in BALB/c mice. The results suggest that the constellation of C4 and U4 among vRNAs may be one of the multigenic determinants of IAV pathogenicity.
Keywords: influenza A virus, pathogenicity, promoter, reverse genetics, viral replication

© 2017 The Korean Society of Veterinary Science.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.