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ORF005L from infectious spleen and kidney necrosis virus is located in the inner mitochondrial membrane and induces apoptosis

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Abstract

Infectious spleen and kidney necrosis virus (ISKNV) belongs to the genus Megalocytivirus in the family Iridoviridae. This virus is the etiological agent of a serious and pandemic disease in fish. Cells infected with ISKNV undergo apoptosis. In this study, the ISKNV ORF005L gene was characterized and functionally investigated. Bioinformatics analysis revealed that the ORF005L protein contains a region similar to the catalytic domain of CTD-like phosphatases. Real-time quantitative-PCR results showed the transcription profile of ORF005L in ISKNV-infected cells; in these cells, ORF005L was initially transcribed at 24 h post-infection. The green fluorescent protein-tagged ORF005L protein was localized in the mitochondria. Sub-mitochondrial fractions were subjected to Western blot, and the results showed that ORF005L was specifically located in the inner membrane of the mitochondria. The ORF005L in fathead minnow cells was transiently expressed, resulting in the decrease in mitochondrial transmembrane potential, which induced cell apoptosis. ORF005L was knocked down by specific dsRNA, thereby significantly reducing the apoptosis of mandarin fish fry-1 cells induced by ISKNV infection. These results indicated that the ORF005L of ISKNV could disrupt mitochondrial function and cause apoptosis. This study may provide novel insights into the pathogenesis of Megalocytivirus infection.

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Acknowledgments

This research was supported by the Natural Science Foundation of China (Grant No. 31330080), the National Basic Research Program of China (973 Program; Grant No. 2012CB114402),the National Natural Science Foundation of China (Young-General Continuous Support Program; Grant No. 31270048), the National Natural Science Foundation for Young Scholars (Grant No. 30901114), the China Postdoctoral Science Special Foundation (Grant No. 201003369), and the Fundamental Research Funds for the Central Universities (Grant No. 11lgpy24).

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Authors and Affiliations

  1. MOE Key Laboratory of Aquatic Product Safety / State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, 510275, People’s Republic of China

    Rui Wang, Yang Yi, Lihong Liu, Yanghong Lu, Shaoping Weng, Jianguo He & Xiaopeng Xu

  2. School of Marine Sciences, Sun Yat-sen University, Guangzhou, 510275, People’s Republic of China

    Jianguo He

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  1. Rui Wang
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  2. Yang Yi
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  3. Lihong Liu
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  4. Yanghong Lu
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  7. Xiaopeng Xu
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Correspondence to Jianguo He or Xiaopeng Xu.

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Rui Wang and Yang Li have contributed equally to this work.

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Fig. S1

Multiple sequence alignment (using Clustal Omega software) of the ISKNV ORF005L protein and its homologs from other iridoviruses. The identical amino acid residues were shaded in black and the similar residues in gray. Proteins analyzed list below: ISKNV ORF005L (Genbank Accession No. NP_612227); Red seabream iridovirus (RSIV) ORF385R (Accession No. BAK14278); Turbot reddish body iridovirus (TRBIV) ORF5 (Accession No. ADE34350); Rock bream iridovirus (RBIV) ORF006L (Accession No. AAT71821); Lymphocystis disease virus-isolate China (LCDV-C) ORF147 (Accession No. YP_073653); Lymphocystis disease virus-1 (LCDV-1) ORF64 (Accession No. NP_078678); Tiger frog virus (TFV) ORF040R (Accession No. ABB92302); and Invertebrate iridescent virus 6 (IIV-6) ORF355R (Accession No. NP_149818). Supplementary material 1 (TIFF 5643 kb)

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Wang, R., Yi, Y., Liu, L. et al. ORF005L from infectious spleen and kidney necrosis virus is located in the inner mitochondrial membrane and induces apoptosis. Virus Genes 49, 269–277 (2014). https://doi.org/10.1007/s11262-014-1088-2

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  • DOI: https://doi.org/10.1007/s11262-014-1088-2

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