title: A gammaherpesvirus complement regulatory protein promotes initiation of infection by activation of protein kinase Akt/PKB.
abstract: BACKGROUND
  Viruses have evolved to evade the host's complement system. The open reading frames 4 (ORF4) of gammaherpesviruses encode homologs of regulators of complement activation (RCA) proteins, which inhibit complement activation at the level of C3 and C4 deposition. Besides complement regulation, these proteins are involved in heparan sulfate and glycosaminoglycan binding, and in case of MHV-68, also in viral DNA synthesis in macrophages.
  
  
  RESULTS
  Here, we made use of MHV-68 to study the role of ORF4 during infection of fibroblasts. While attachment and penetration of virions lacking the RCA protein were not affected, we observed a delayed delivery of the viral genome to the nucleus of infected cells. Analysis of the phosphorylation status of a variety of kinases revealed a significant reduction in phosphorylation of the protein kinase Akt in cells infected with ORF4 mutant virus, when compared to cells infected with wt virus. Consistent with a role of Akt activation in initial stages of infection, inhibition of Akt signaling in wt virus infected cells resulted in a phenotype resembling the phenotype of the ORF4 mutant virus, and activation of Akt by addition of insulin partially reversed the phenotype of the ORF4 mutant virus. Importantly, the homologous ORF4 of KSHV was able to rescue the phenotype of the MHV-68 ORF4 mutant, indicating that ORF4 is functionally conserved and that ORF4 of KSHV might have a similar function in infection initiation.
  
  
  CONCLUSIONS
  In summary, our studies demonstrate that ORF4 contributes to efficient infection by activation of the protein kinase Akt and thus reveal a novel function of a gammaherpesvirus RCA protein.
authors.affiliation: The Institute of Molecular Immunology, Clinical Cooperation Group Hematopoietic Cell Transplantation, Helmholtz Zentrum München-German Research Center for Environmental Health, Munich, Germany.
authors.forename: Beatrix
authors.initials: B
authors.lastname: Steer
authors.forename: Barbara
authors.initials: B
authors.lastname: Adler
authors.forename: Stipan
authors.initials: S
authors.lastname: Jonjic
authors.forename: James P
authors.initials: JP
authors.lastname: Stewart
authors.forename: Heiko
authors.initials: H
authors.lastname: Adler
mesh_terms: D000818:Animals; D005434:Flow Cytometry; D018142:Gammaherpesvirinae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005816:Genetic Complementation Test; D007328:Insulin / Q000494:pharmacology; D007700:Kinetics; D051379:Mice; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D016366:Open Reading Frames / Q000235:genetics; D010766:Phosphorylation / Q000187:drug effects; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012263:Ribonucleosides / Q000494:pharmacology; D015398:Signal Transduction; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology*
chemical_list: D007328:Insulin; D012263:Ribonucleosides; D014764:Viral Proteins; C023764:triciribine; D051057:Proto-Oncogene Proteins c-akt