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Disruption of PML Nuclear Bodies Is Mediated by ORF61 SUMO-Interacting Motifs and Required for Varicella-Zoster Virus Pathogenesis in Skin

By Li Wang, Stefan L. Oliver, Marvin Sommer, Jaya Rajamani, Mike Reichelt and Ann M. Arvin


Promyelocytic leukemia protein (PML) has antiviral functions and many viruses encode gene products that disrupt PML nuclear bodies (PML NBs). However, evidence of the relevance of PML NB modification for viral pathogenesis is limited and little is known about viral gene functions required for PML NB disruption in infected cells in vivo. Varicella-zoster virus (VZV) is a human alphaherpesvirus that causes cutaneous lesions during primary and recurrent infection. Here we show that VZV disrupts PML NBs in infected cells in human skin xenografts in SCID mice and that the disruption is achieved by open reading frame 61 (ORF61) protein via its SUMO-interacting motifs (SIMs). Three conserved SIMs mediated ORF61 binding to SUMO1 and were required for ORF61 association with and disruption of PML NBs. Mutation of the ORF61 SIMs in the VZV genome showed that these motifs were necessary for PML NB dispersal in VZV-infected cells in vitro. In vivo, PML NBs were highly abundant, especially in basal layer cells of uninfected skin, whereas their frequency was significantly decreased in VZV-infected cells. In contrast, mutation of the ORF61 SIMs reduced ORF61 association with PML NBs, most PML NBs remained intact and importantly, viral replication in skin was severely impaired. The ORF61 SIM mutant virus failed to cause the typical VZV lesions that penetrate across the basement membrane into the dermis and viral spread in the epidermis was limited. These experiments indicate that VZV pathogenesis in skin depends upon the ORF61-mediated disruption of PML NBs and that the ORF61 SUMO-binding function is necessary for this effect. More broadly, our study elucidates the importance of PML NBs for the innate control of a viral pathogen during infection of differentiated cells within their tissue microenvironment in vivo and the requirement for a viral protein with SUMO-binding capacity to counteract this intrinsic barrier

Topics: Research Article
Publisher: Public Library of Science
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Provided by: PubMed Central

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  1. (1998). A novel nuclear substructure, ND10: distribution in normal and neoplastic human tissues.
  2. (2000). Alphaherpesvirus proteins related to herpes simplex virus type 1 ICP0 affect cellular structures and proteins.
  3. (2007). Cellular and viral factors regulate the varicella-zoster virus gE promoter during viral replication.
  4. (1995). Characterization of a new monoclonal antibody (PG-M3) directed against the aminoterminal portion of the PML gene product: immunocytochemical evidence for high expression of PML proteins on activated macrophages, endothelial cells, and epithelia.
  5. (2009). Components of nuclear domain 10 bodies regulate varicella-zoster virus replication.
  6. (2007). Conserved function of RNF4 family proteins in eukaryotes: targeting a ubiquitin ligase to SUMOylated proteins.
  7. (2003). Construction of varicella-zoster virus recombinants from parent Oka cosmids and demonstration that ORF65 protein is dispensable for infection of human skin and T cells in the SCID-hu mouse model.
  8. (2000). Covalent modification of p73a by SUMO-1. Two-hybrid screening with p73 identifies novel SUMO-1-interacting proteins and a SUMO-1 interaction motif.
  9. (1999). de The H
  10. (2005). Defining the SUMO-modified proteome by multiple approaches in Saccharomyces cerevisiae.
  11. (2006). Dikic I
  12. (2002). Effects of promyelocytic leukemia protein on virus-host balance.
  13. (2011). Entrapment of viral capsids in nuclear PML cages is an intrinsic host defense against a neurotropic human herpesvirus.
  14. (1988). Expression of the proliferation markers Ki67 and transferrin receptor by human trophoblast populations.
  15. (2008). Functions of Varicella-zoster virus ORF23 capsid protein in viral replication and the pathogenesis of skin infection.
  16. (2010). Genome-wide mutagenesis reveals that ORF7 is a novel VZV skin tropic factor.
  17. (2002). Herpes simplex virus 1-infected cell protein 0 contains two E3 ubiquitin ligase sites specific for different E2 ubiquitin-conjugating enzymes.
  18. (2010). Herpes simplex virus tegument ICP0 is capsid associated, and its E3 ubiquitin ligase domain is important for incorporation into virions.
  19. (2002). Herpes simplex virus type 1 immediateearly protein ICP0 and is isolated RING finger domain act as ubiquitin E3 ligases in vitro.
  20. (2007). Herpes simplex virus. Fields virology, 5th ed, eds Knipe DM, Howley PM,
  21. (2006). Inhibition of the NF-kappaB pathway by varicellazoster virus in vitro and in human epidermal cells in vivo.
  22. (2006). Investigations of the pathogenesis of Varicella zoster virus infection in the SCIDhu mouse model.
  23. (2009). Kaposi’s sarcoma-associated herpesvirus protein LANA2 disrupts PML oncogenic domains and inhibits PML-mediated transcriptional repression of the survivin gene.
  24. (1999). MDM2 interacts with MDMX through their RING finger domains.
  25. (2006). Nuclear and chromatin reorganization in the MHC-Oct3/4 locus at developmental phases of embryonic stem cell differentiation.
  26. (1995). Orr A
  27. (2007). PML and PML nuclear bodies: implications in antiviral defence.
  28. (2006). PML contributes to a cellular mechanism of repression of herpes simplex virus type 1 infection that is inactivated by ICP0.
  29. (1999). PML is critical for ND10 formation and recruits the PML-interacting protein daxx to this nuclear structure when modified by SUMO-1.
  30. (2003). Promyelocytic leukemia protein mediates interferon-based anti-herpes simplex virus 1 effects.
  31. (2009). Regulation of the ORF61 promoter and ORF61 functions in varicella-zoster virus replication and pathogenesis.
  32. (2008). RNF4 is a poly-SUMO-specific E3 ubiquitin ligase required for arsenic-induced PML degradation.
  33. (2006). Role of SUMOinteracting motif in Daxx SUMO modification, subnuclear localization, and repression of sumoylated transcription factors.
  34. (2005). Small ubiquitin-like modifier (SUMO) recognition of a SUMO binding motif: a reversal of the bound orientation.
  35. (2007). Structure, dynamics and functions of promyelocytic leukaemia nuclear bodies.
  36. (2009). SUMO interaction motifs in Sizn1 are required for promyelocytic leukemia protein nuclear body localization and for transcriptional activation.
  37. (1995). The acute promyelocytic leukaemia-associated PML gene is induced by interferon.
  38. (2004). The herpes simplex virus ICP0 RING finger domain inhibits IRF3- and IRF7-mediated activation of interferon-stimulated genes.
  39. (2006). The mechanisms of PML-nuclear body formation.
  40. (1994). The nuclear location of PML, a cellular member of the C3HC4 zinc-binding domain protein family, is rearranged during herpes simplex virus infection by the C3HC4 viral protein ICP0.
  41. (2009). The replication cycle of varicella-zoster virus: analysis of the kinetics of viral protein expression, genome synthesis, and virion assembly at the single-cell level.
  42. (1994). The RING finger domain of the varicella-zoster virus open reading frame 61 protein is required for its transregulatory functions.
  43. (1995). Tropism of varicellazoster virus for human CD4+ and CD8+ T lymphocytes and epidermal cells in SCID-hu mice.
  44. (1993). Varicella-zoster virus (VZV) open reading frame 61 protein transactivates VZV gene promoters and enhances the infectivity of VZV DNA.
  45. (2005). Varicella-zoster virus infection of human dorsal root ganglia in vivo.
  46. (1998). Varicella-zoster virus ORF61 deletion mutants replicate in cell culture, but a mutant with stop codons in ORF61 reverts to wildtype virus.
  47. (2007). Varicella-zoster virus replication, pathogenesis, and management. Fields virology, 5th ed, eds Knipe DM, Howley PM,
  48. (2004). Varicella-zoster virus transfer to skin by T Cells and modulation of viral replication by epidermal cell interferon-alpha.
  49. (2010). Varicellazoster virus immediate-early protein 62 blocks interferon regulatory factor 3 (IRF3) phosphorylation at key serine residues: a novel mechanism of IRF3 inhibition among herpesviruses.