1887

Abstract

Human cytomegalovirus UL99-encoded tegument protein pp28 contains a 16 aa acidic cluster that is required for pp28 trafficking to the assembly compartment (AC) and the virus assembly. However, functional signals within the acidic cluster of pp28 remain undefined. Here, we demonstrated that an acidic cluster rather than specific sorting signals was required for trafficking to the AC. Recombinant viruses with chimeric pp28 proteins expressing non-native acidic clusters exhibited delayed viral growth kinetics and decreased production of infectious virus, indicating that the native acidic cluster of pp28 was essential for wild-type virus assembly. These results suggested that the acidic cluster of pp28 has distinct functional domains required for trafficking and for efficient virus assembly. The first half (aa 44–50) of the acidic cluster was sufficient for pp28 trafficking, whereas the native acidic cluster consisting of aa 51–59 was required for the assembly of wild-type levels of infectious virus.

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2016-10-13
2024-11-03
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References

  1. Andreoni M., Faircloth M., Vugler L., Britt W. J. 1989; A rapid microneutralization assay for the measurement of neutralizing antibody reactive with human cytomegalovirus. J Virol Methods 23:157–167 [View Article][PubMed]
    [Google Scholar]
  2. Bonifacino J. S., Traub L. M. 2003; Signals for sorting of transmembrane proteins to endosomes and lysosomes. Annu Rev Biochem 72:395–447 [View Article][PubMed]
    [Google Scholar]
  3. Borst E. M., Hahn G., Koszinowski U. H., Messerle M. 1999; Cloning of the human cytomegalovirus (HCMV) genome as an infectious bacterial artificial chromosome in Escherichia coli: a new approach for construction of HCMV mutants. J Virol 73:8320–8329
    [Google Scholar]
  4. Britt W. J., Jarvis M., Seo J. Y., Drummond D., Nelson J. 2004; Rapid genetic engineering of human cytomegalovirus by using a lambda phage linear recombination system: demonstration that pp28 (UL99) is essential for production of infectious virus. J Virol 78:539–543 [View Article][PubMed]
    [Google Scholar]
  5. Chee M. S., Bankier A. T., Beck S., Bohni R., Brown C. M., Cerny R., Horsnell T., Hutchison C. A., Kouzarides T. et al. 1990; Analysis of the protein-coding content of the sequence of human cytomegalovirus strain AD169. Curr Top Microbiol Immunol 154:125–170[PubMed]
    [Google Scholar]
  6. Crump C. M., Xiang Y., Thomas L., Gu F., Austin C., Tooze S. A., Thomas G. 2001; PACS-1 binding to adaptors is required for acidic cluster motif-mediated protein traffic. EMBO J 20:2191–2201 [View Article][PubMed]
    [Google Scholar]
  7. Jones T. R., Lee S. W. 2004; An acidic cluster of human cytomegalovirus UL99 tegument protein is required for trafficking and function. J Virol 78:1488–1502 [View Article][PubMed]
    [Google Scholar]
  8. Mach M., Kropff B., Kryzaniak M., Britt W. 2005; Complex formation by glycoproteins M and N of human cytomegalovirus: structural and functional aspects. J Virol 79:2160–2170 [View Article][PubMed]
    [Google Scholar]
  9. Meyer H., Bankier A. T., Landini M. P., Brown C. M., Barrell B. G., Rüger B., Mach M. 1988; Identification and procaryotic expression of the gene coding for the highly immunogenic 28-kilodalton structural phosphoprotein (pp28) of human cytomegalovirus. J Virol 62:2243–2250[PubMed]
    [Google Scholar]
  10. Misra S., Puertollano R., Kato Y., Bonifacino J. S., Hurley J. H. 2002; Structural basis for acidic-cluster-dileucine sorting-signal recognition by VHS domains. Nature 415:933–937 [View Article][PubMed]
    [Google Scholar]
  11. Molloy S. S., Anderson E. D., Jean F., Thomas G. 1999; Bi-cycling the furin pathway: from TGN localization to pathogen activation and embryogenesis. Trends Cell Biol 9:28–35 [View Article]
    [Google Scholar]
  12. Phillips S. L., Bresnahan W. A. 2012; The human cytomegalovirus (HCMV) tegument protein UL94 is essential for secondary envelopment of HCMV virions. J Virol 86:2523–2532 [View Article][PubMed]
    [Google Scholar]
  13. Phillips S. L., Cygnar D., Thomas A., Bresnahan W. A. 2012; Interaction between the human cytomegalovirus tegument proteins UL94 and UL99 is essential for virus replication. J Virol 86:9995–10005 [View Article][PubMed]
    [Google Scholar]
  14. Robinson M. S. 2004; Adaptable adaptors for coated vesicles. Trends Cell Biol 14:167–174 [View Article][PubMed]
    [Google Scholar]
  15. Sanchez V., Greis K. D., Sztul E., Britt W. J. 2000a; Accumulation of virion tegument and envelope proteins in a stable cytoplasmic compartment during human cytomegalovirus replication: characterization of a potential site of virus assembly. J Virol 74:975–986 [View Article][PubMed]
    [Google Scholar]
  16. Sanchez V., Sztul E., Britt W. J. 2000b; Human cytomegalovirus pp28 (UL99) localizes to a cytoplasmic compartment which overlaps the endoplasmic reticulum-Golgi-intermediate compartment. J Virol 74:3842–3851 [View Article][PubMed]
    [Google Scholar]
  17. Seo J. Y., Britt W. J. 2006; Sequence requirements for localization of human cytomegalovirus tegument protein pp28 to the virus assembly compartment and for assembly of infectious virus. J Virol 80:5611–5626 [View Article][PubMed]
    [Google Scholar]
  18. Seo J. Y., Britt W. J. 2007; Cytoplasmic envelopment of human cytomegalovirus requires the postlocalization function of tegument protein pp28 within the assembly compartment. J Virol 81:6536–6547 [View Article][PubMed]
    [Google Scholar]
  19. Seo J. Y., Britt W. J. 2008; Multimerization of tegument protein pp28 within the assembly compartment is required for cytoplasmic envelopment of human cytomegalovirus. J Virol 82:6272–6287 [View Article][PubMed]
    [Google Scholar]
  20. Shiba T., Takatsu H., Nogi T., Matsugaki N., Kawasaki M., Igarashi N., Suzuki M., Kato R., Earnest T. et al. 2002; Structural basis for recognition of acidic-cluster dileucine sequence by GGA1. Nature 415:937–941 [View Article][PubMed]
    [Google Scholar]
  21. Silva M. C., Yu Q. C., Enquist L., Shenk T. 2003; Human cytomegalovirus UL99-encoded pp28 is required for the cytoplasmic envelopment of tegument-associated capsids. J Virol 77:10594–10605 [View Article][PubMed]
    [Google Scholar]
  22. Smith J. D., De Harven E. 1973; Herpes simplex virus and human cytomegalovirus replication in WI-38 cells. I. Sequence of viral replication. J Virol 12:919–930[PubMed]
    [Google Scholar]
  23. Warming S., Costantino N., Court D. L., Jenkins N. A., Copeland N. G. 2005; Simple and highly efficient BAC recombineering using galK selection. Nucleic Acids Res 33:e36 [View Article][PubMed]
    [Google Scholar]
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