Mechanistic Insight into the Host Transcription Inhibition Function of Rift Valley Fever Virus NSs and Its Importance in Virulence

Rift Valley fever virus (RVFV), a member of the genus Phlebovirus within the family Bunyaviridae, causes periodic outbreaks in livestocks and humans in countries of the African continent and Middle East. RVFV NSs protein, a nonstructural protein, is a major virulence factor that exhibits several important biological properties. These include suppression of general transcription, inhibition of IFN-β promoter induction and degradation of double-stranded RNA-dependent protein kinase R. Although each of these biological functions of NSs are considered important for countering the antiviral response in the host, the individual contributions of these functions towards RVFV virulence remains unclear. To examine this, we generated two RVFV MP-12 strain-derived mutant viruses. Each carried mutations in NSs that specifically targeted its general transcription inhibition function without affecting its ability to degrade PKR and inhibit IFN-β promoter induction, through its interaction with Sin3-associated protein 30, a part of the repressor complex at the IFN-β promoter. Using these mutant viruses, we have dissected the transcription inhibition function of NSs and examined its importance in RVFV virulence. Both NSs mutant viruses exhibited a differentially impaired ability to inhibit host transcription when compared with MP-12. It has been reported that NSs suppresses general transcription by interfering with the formation of the transcription factor IIH complex, through the degradation of the p62 subunit and sequestration of the p44 subunit. Our study results lead us to suggest that the ability of NSs to induce p62 degradation is the major contributor to its general transcription inhibition property, whereas its interaction with p44 may not play a significant role in this function. Importantly, RVFV MP-12-NSs mutant viruses with an impaired general transcription inhibition function showed a reduced cytotoxicity in cell culture and attenuated virulence in young mice, compared with its parental virus MP-12, highlighting the contribution of NSs-mediated general transcription inhibition towards RVFV virulence.
Author Summary Rift Valley fever virus (RVFV) has a significant impact on the livestock industry because of its high mortality rate in young ruminants and causation of a high abortion rate in pregnant animals. Human RVFV infections generally manifest as self-limiting and non-fatal illnesses. However, a small percentage of patients develop encephalitis, vision loss and hemorrhagic fever with a high mortality rate. Currently, there is no commercially available vaccine for human use or effective antiviral drug for RVFV treatment. The non-structural protein NSs is a major virulence factor of RVFV, which mediates suppression of host general transcription, inhibition of IFN-β transcription and degradation of PKR, to block host antiviral responses. To examine the contribution of host transcription inhibition to RVFV virulence, we generated RVFV MP-12 strain-derived mutants that have attenuated inhibitory activity on host transcription due to amino acid mutations in NSs. The mutant viruses showed attenuated cytotoxicity in cell culture and attenuated virulence in young mice, demonstrating the contribution of NSs-mediated host transcription inhibition to the virulence of RVFV.