Abstract
Rabbit hemorrhagic disease (RHD) is a highly contagious disease caused by rabbit hemorrhagic disease virus (RHDV). Previous research has shown that RHDV induces apoptosis in numerous cell types, although the molecular mechanisms underlying the apoptosis induced by RHDV are not well understood. One possible factor is non-structural protein 6 (NSP6), a 3C-like protease that plays an important role in processing viral polyprotein precursors into mature non-structural proteins. To fully establish a role for NSP6, the present study examined the effects of ectopic expression of the protein in rabbit (RK13) and human (HeLa and HepG2) cells. We found that NSP6 suppressed cell viability and promoted apoptosis in all three cell types in a dose-dependent manner. We also identified increased caspase-3, -8, and -9 activities in RK13 cell, and an increased Bax to Bcl2 mRNA ratio. Mechanistically, the ability of NSP6 to induce apoptosis was impaired by mutation of the catalytic His27 residue. Our study has shown that RHDV NSP6 can induce apoptosis in host cells and is likely an important contributor to RHDV-induced apoptosis and pathogenesis.
Highlights
Rabbit hemorrhagic disease (RHD) is a highly contagious disease characterized by acute liver damage and disseminated intravascular coagulation (DIC) (Alonso et al, 1998; Jung et al, 2000)
In trypan blue dye exclusion test, the viable cell rate of RK13 cells transfected with non-structural protein 6 (NSP6) reduced to 72, 66 and 65% at 24, 48 and 72 hpt (Figure 2B), so there is a slight discrepancy between the results of MTT assay and trypan blue dye exclusion test
rabbit hemorrhagic disease virus (RHDV) research was limited due to the lack of a stable cell system to culture the virus in vitro. It has remained unclear which viral components contribute to RHDV-induced apoptosis
Summary
Rabbit hemorrhagic disease (RHD) is a highly contagious disease characterized by acute liver damage and disseminated intravascular coagulation (DIC) (Alonso et al, 1998; Jung et al, 2000). The first outbreak of RHD was reported in 1984 in the Jiangsu Province of China (Zhu et al, 2016) and has quickly spread to most parts of the world (Calvete et al, 2018; Dalton et al, 2018). This has led to the deaths of millions of rabbits and hares, representing a serious threat to their populations. RHDV contains both genomic RNA (gRNA) and additional subgenomic RNA sequences (sgRNA). Of the non-structural proteins, NSP6 is known
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