title: TRAF2 facilitates vaccinia virus replication by promoting rapid virus entry.
abstract: Tumor necrosis factor receptor (TNFR)-associated factor 2 (TRAF2) is a pivotal intracellular mediator of signaling pathways downstream of TNFR1 and -2 with known pro- and antiviral effects. We investigated its role in the replication of the prototype poxvirus vaccinia virus (VACV). Loss of TRAF2 expression, either through small interfering RNA treatment of HeLa cells or through genetic knockout in murine embryonic fibroblasts (MEFs), led to significant reductions in VACV growth following low-multiplicity infection. In single-cycle infections, there was delayed production of both early and late VACV proteins as well as accelerated virus-induced alterations to cell morphology, indicating that TRAF2 influences early stages of virus replication. Consistent with an early role, uncoating assays showed normal virus attachment but delayed virus entry in the absence of TRAF2. Although alterations to c-Jun N-terminal kinase (JNK) signaling were apparent in VACV-infected TRAF2(-/-) MEFs, treatment of wild-type cells with a JNK inhibitor did not affect virus entry. Instead, treatment with an inhibitor of endosomal acidification greatly reduced virus entry into TRAF2(-/-) MEFs, suggesting that VACV is reliant on the endosomal route of entry in the absence of TRAF2. Thus, TRAF2 is a proviral factor for VACV that plays a role in promoting efficient viral entry, most likely via the plasma membrane.
  
  
  OBJECTIVE
  Tumor necrosis factor receptor-associated factors (TRAFs) are key facilitators of intracellular signaling with roles in innate and adaptive immunity and stress responses. We have discovered that TRAF2 is a proviral factor in vaccinia virus replication in both HeLa cells and mouse embryonic fibroblasts and that its influence is exercised through promotion of efficient virus entry.
authors.affiliation: The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Ismar R
authors.initials: IR
authors.lastname: Haga
authors.forename: Tali
authors.initials: T
authors.lastname: Pechenick Jowers
authors.forename: Samantha J
authors.initials: SJ
authors.lastname: Griffiths
authors.forename: Juergen
authors.initials: J
authors.lastname: Haas
authors.forename: Philippa M
authors.initials: PM
authors.lastname: Beard
mesh_terms: D000818:Animals; D002460:Cell Line; D004847:Epithelial Cells / Q000821:virology; D005347:Fibroblasts / Q000821:virology; D055785:Gene Knockdown Techniques; D055786:Gene Knockout Techniques; D054884:Host-Pathogen Interactions; D006801:Humans; D051379:Mice; D047992:TNF Receptor-Associated Factor 2 / Q000235:genetics / Q000378:metabolism*; D014616:Vaccinia virus / Q000502:physiology*; D053586:Virus Internalization
chemical_list: D047992:TNF Receptor-Associated Factor 2