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.

We also used RIG-I exclusion and viral protein/RNA markers to examine the cellular location of viral protein production. A spatial separation of viral RNA replication and translation has been proposed based on evidence that various positive-strand RNA virus replication complexes are composed of ribosome-free membranes and observations that these processes interfere with one another [6, 11-13, 48, 57, 58] . Here, we show that bulk ribosomes have a similar subcellular localization to that of RIG-I in HCV-infected cells, supporting the view that a physical separation exists between translation and genome replication/assembly (Fig 5 and S3 Fig) . Consistent with this, a portion of the viral positive-strand RNA is found in regions containing RIG-I and ribosomes (Fig 3B) . These observations led us to conclude that viral RNA destined for translation may exit the MW. Such a mechanism would require that some fraction of newly synthesized viral RNA be exported from the MW for translation, while another pool is retained in the MW and targeted to viral assembly centers. As discussed below, we envisage that components of the nuclear transport machinery may contribute to these targeting events.