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Identification of RNA regions that determine temperature sensitivities in betanodaviruses

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Abstract

Betanodaviruses, the causative agents of viral nervous necrosis in marine fish, have bipartite positive-sense RNA genomes. The larger genomic segment, RNA1 (~3.1 kb), encodes an RNA-dependent RNA polymerase (protein A), and the smaller genomic segment RNA2 (~1.4 kb) codes for the coat protein. These viruses can be classified into four genotypes, designated striped jack nervous necrosis virus (SJNNV), redspotted grouper nervous necrosis virus (RGNNV), tiger puffer nervous necrosis virus (TPNNV), and barfin flounder nervous necrosis virus (BFNNV), based on similarities in their partial RNA2 sequences. The optimal temperatures for the growth of these viruses are 20–25°C (SJNNV), 25–30°C (RGNNV), 20°C (TPNNV), and 15–20°C (BFNNV). However, little is known about the mechanisms underlying the temperature sensitivity of these viruses. We first constructed two reassortants between SJNNV and RGNNV to test their temperature sensitivity. The levels of viral growth and RNA replication of these reassortants and parental viruses in cultured fish cells were similar at 25°C. However, the levels of all of the viruses but RGNNV were markedly reduced at 30°C. These results indicate that both RNA1 and RNA2 control the temperature sensitivity of betanodaviruses by modulating RNA replication or earlier viral growth processes. We then constructed ten mutated RGNNVs, the RNA1 segments of which were chimeric between SJNNV and RGNNV, and showed that only chimeric viruses bearing the RGNNV RNA1 region, encoding amino acid residues 1–445, grew similarly to the parental RGNNV at 30°C. This portion of protein A is known to serve as a mitochondrial-targeting signal rather than functioning as an enzymatic domain.

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Acknowledgments

This work was supported in part by grants-in-aid for Scientific Research (18580185, 20380111) from the Ministry of Education, Culture, Sports, Science and Technology, Japan and a grant-in-aid for Scientific Research (18076) from the Ministry of Agriculture, Forestry and Fisheries of Japan.

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Authors and Affiliations

  1. Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima, 739-8528, Japan

    Naomi Hata, Yasushi Okinaka, Yasuhiko Kawato & Toshihiro Nakai

  2. Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, PE, C1A 4P3, Canada

    Tokinori Iwamoto

  3. National Research Institute of Aquaculture, Fisheries Research Agency, Saiki, 879-2602, Japan

    Koh-Ichiro Mori

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  1. Naomi Hata
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  2. Yasushi Okinaka
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Correspondence to Yasushi Okinaka.

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Hata, N., Okinaka, Y., Iwamoto, T. et al. Identification of RNA regions that determine temperature sensitivities in betanodaviruses. Arch Virol 155, 1597–1606 (2010). https://doi.org/10.1007/s00705-010-0736-7

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