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Article

Construction of an Influenza D Virus with an Eight-Segmented Genome

Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan
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Academic Editor: Stacey Schultz-Cherry
Viruses 2021, 13(11), 2166; https://doi.org/10.3390/v13112166
Received: 29 September 2021 / Revised: 22 October 2021 / Accepted: 25 October 2021 / Published: 27 October 2021
(This article belongs to the Section Animal Viruses)
Influenza D virus (IDV) may cause the bovine respiratory disease complex, which is the most common and costly disease affecting the cattle industry. Previously, we revealed that eight segments could be actively packaged in its single virion, suggesting that IDV with the seven-segmented genome shows an agnostic genome packaging mechanism. Herein, we engineered an eight-segmented recombinant IDV in which the NS1 or NS2 genes were separated from NS segment into independent segments (NS1 or NS2 segments, respectively), leading to monocistronic translation of each NS protein. We constructed two plasmids: one for the viral RNA (vRNA)-synthesis of the NS1 segment with a silent mutation at the splicing acceptor site, which controls NS2 transcription in the NS segment; and another for the RNA synthesis of the NS2 segment, with deletion of the intron in the NS segment. These plasmids and six other vRNA-synthesis plasmids were used to fabricate an infectious eight-segmented IDV via reverse genetics. This system enables analysis of the functions of NS1 or NS2. We tested the requirement of the N-terminal overlapping region (NOR) in these proteins for viral infectivity. We rescued a virus with NOR-deleted NS2 protein, which displayed a growth rate equivalent to that of the eight-segmented virus with intact NS2. Thus, the NOR may not influence viral growth. In contrast, a virus with NOR-deleted NS1 protein could not be rescued. These results indicate that the eight-segmented rescue system of IDV may provide an alternative method to analyze viral proteins at the molecular level. View Full-Text
Keywords: bovine respiratory disease complex; influenza D virus; mutant; recombinant virus; reverse genetics; nonstructural protein bovine respiratory disease complex; influenza D virus; mutant; recombinant virus; reverse genetics; nonstructural protein
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MDPI and ACS Style

Ishida, H.; Murakami, S.; Kamiki, H.; Matsugo, H.; Katayama, M.; Sekine, W.; Ohira, K.; Takenaka-Uema, A.; Horimoto, T. Construction of an Influenza D Virus with an Eight-Segmented Genome. Viruses 2021, 13, 2166. https://doi.org/10.3390/v13112166

AMA Style

Ishida H, Murakami S, Kamiki H, Matsugo H, Katayama M, Sekine W, Ohira K, Takenaka-Uema A, Horimoto T. Construction of an Influenza D Virus with an Eight-Segmented Genome. Viruses. 2021; 13(11):2166. https://doi.org/10.3390/v13112166

Chicago/Turabian Style

Ishida, Hiroho, Shin Murakami, Haruhiko Kamiki, Hiromichi Matsugo, Misa Katayama, Wataru Sekine, Kosuke Ohira, Akiko Takenaka-Uema, and Taisuke Horimoto. 2021. "Construction of an Influenza D Virus with an Eight-Segmented Genome" Viruses 13, no. 11: 2166. https://doi.org/10.3390/v13112166

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