The Hepatitis C Virus-Induced Membranous Web and Associated Nuclear Transport Machinery Limit Access of Pattern Recognition Receptors to Viral Replication Sites

Neufeldt, Christopher J.; Joyce, Michael A.; Van Buuren, Nicholas; Levin, Aviad; Kirkegaard, Karla; Gale Jr., Michael; Tyrrell, D. Lorne J.; Wozniak, Richard W.

In addition to its role in establishing distinct compartments for viral replication and assembly within its boundaries, the MW has also been suggested to function more broadly as physical barrier between these processes and the surrounding cytoplasm. This physical separation has been proposed to suppress the innate immune response, at least in part, by limiting access of RLRs to viral RNA (reviewed in [24, 72] ). In this study, we provide evidence that cytoplasmic RIG-I and MDA5 are restricted from entering regions of the cytoplasm occupied by various HCV proteins that serve as molecular markers for the MW (Fig 1 and S1B Fig) . Moreover, probes specific for positive-or negative-strand viral RNA reveal that RIG-I is excluded from regions of the MW containing both replication and assembly complexes (Figs 1 and 3 and S1A Fig) . These results provided direct experimental evidence that the MW can function to limit access of innate immune receptors to viral RNA.