Skip to main content
NCBI home page
Wiley - PMC COVID-19 Collection logoLink to Wiley - PMC COVID-19 Collection
. 2006 May 18;16(3):187–202. doi: 10.1002/rmv.499

Vaccinia virus proteolysis—a review

Chelsea M Byrd 1, Dennis E Hruby 1,2,✉
PMCID: PMC7169229  PMID: 16710840

Abstract

It is well known that viruses, as obligate intracellular parasites, must use their hosts' metabolic machinery in order to replicate their genomes and form infectious progeny virions. What is less well known are the details of how viruses make sure that once all the necessary proteins are made, that they assume the correct configuration at the proper time in order to catalyse the efficient assembly of infectious virions. One of the methods employed by viruses to regulate this process is the proteolytic cleavage of viral proteins. Over the past several decades, studies in numerous laboratories have demonstrated that morphogenic proteolysis plays a major and essential role during the assembly and maturation of infectious poxvirus virions. In this review we describe the history of vaccinia virus proteolysis as a prototypic viral system. Copyright © 2006 John Wiley & Sons, Ltd.

REFERENCES

  • 1. Linderstrom‐Lang KU, Ottesen M. Formation of plakalbumin from ovalbumin. Comp Rend Trav Lab Carlsberg 1949; 26: 403–412. [Google Scholar]
  • 2. Polgar L. Mechanisms of protease action. CRC Press: Boca Raton, Florida, 1989. [Google Scholar]
  • 3. Dougherty WG, Semler BL. Expression of virus‐encoded proteinases: functional and structural similarities with cellular enzymes. Microbiol Rev 1993; 57: 781–822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4. Polgar L. Structure and function of serine proteinases In New Comprehensive Biochemistry (Vol. 16) Brocklehurst ANaK. (ed.). Elsevier: Amsterdam, 1987. [Google Scholar]
  • 5. Loeb DD, Hutchison CA, Edgell MH, Farmerie WG, Swanstrom R. Mutational analysis of human immunodeficiency virus type 1 protease suggests functional homology with aspartic proteinases. J Virol 1989; 63: 111–121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6. Wright HT. Secondary and conformational specificities of trypsin and chymotrypsin. Eur J Biochem 1977; 73: 567–578. [DOI] [PubMed] [Google Scholar]
  • 7. Neurath H. Proteolytic processing and physiological regulation. Trends Biochem Sci 1989; 14: 268–271. [DOI] [PubMed] [Google Scholar]
  • 8. Horl WH, Heidland AH. Proteases: potential role in health and disease. Adv Exp Med Biol 1982; 167: 1–591. [PubMed] [Google Scholar]
  • 9. Reich E, Rifkin DB, Shaw E. (eds). Proteases and Biological control. Cold Spring Harbor Laboratory Press: New York, 1975. [Google Scholar]
  • 10. Schlesinger S, Schlesinger MJ. Replication of togaviridae and flaviviridae In Virology, 2nd edn, Vol. 1, Fields BN, Knipe DM, Chanock RM, Hirsch MS, Melnick JL, Monath TP, Roizrnan B. (eds). Ravens Press: New York, 1990; 697–710. [Google Scholar]
  • 11. Hellen CU, Wimmer E. The role of proteolytic processing in the morphogenesis of virus particles. Experientia 1992; 48: 201–215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12. Kay J, Dunn BM. Viral proteinases: weakness in strength. Biochim Biophys Acta 1990; 1048: 1–18. [DOI] [PubMed] [Google Scholar]
  • 13. Krausslich HG, Wimmer E. Viral proteinases. Annu Rev Biochem 1988; 57: 701–754. [DOI] [PubMed] [Google Scholar]
  • 14. Stewart L, Schatz G, Vogt VM. Properties of avian retrovirus particles defective in viral protease. J Virol 1990; 64: 5076–5092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15. Hershko A, Fry M. Post‐translational cleavage of polypeptide chains: role in assembly. Annu Rev Biochem 1975; 44: 775–797. [DOI] [PubMed] [Google Scholar]
  • 16. Lee P, Hruby DE. Trans processing of vaccinia virus core proteins. J Virol 1993; 67: 4252–4263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17. Navia MA, McKeever BM. A role for the aspartyl protease from the human immunodeficiency virus type 1 (HIV‐1) in the orchestration of virus assembly. Ann N Y Acad Sci 1990; 616: 73–85. [DOI] [PubMed] [Google Scholar]
  • 18. Pelham HR. Translation of encephalomyocarditis virus RNA in vitro yields an active proteolytic processing enzyme. Eur J Biochem 1978; 85: 457–462. [DOI] [PubMed] [Google Scholar]
  • 19. McGrath WJ, Abola AP, Toledo DL, Brown MT, Mangel WF. Characterization of human adenovirus proteinase activity in disrupted virus particles. Virology 1996; 217: 131–138. [DOI] [PubMed] [Google Scholar]
  • 20. Andres G, Alejo A, Simon‐Mateo C, Salas M. African swine fever virus protease, a new viral member of the SUMO‐1‐specific protease family. J Biol Chem 2001; 276: 780–787. [DOI] [PubMed] [Google Scholar]
  • 21. Webster A, Hay RT, Kemp G. The adenovirus protease is activated by a virus‐coded disulphide‐linked peptide. Cell 1993; 72: 97–104. [DOI] [PubMed] [Google Scholar]
  • 22. Mangel WF, McGrath WJ, Toledo DL, Anderson CW. Viral DNA and a viral peptide can act as cofactors of adenovirus virion proteinase activity. Nature 1993; 361: 274–275. [DOI] [PubMed] [Google Scholar]
  • 23. Choi HK, Tong L, Minor W, et al Structure of Sindbis virus core protein reveals a chymotrypsin‐like serine proteinase and the organization of the virion. Nature 1991; 354: 37–43. [DOI] [PubMed] [Google Scholar]
  • 24. Hruby DE, Guarino LA, Kates JR. Vaccinia virus replication. I. Requirement for the host‐cell nucleus. J Virol 1979; 29: 705–715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. Moss B. Poxviridae and their replication In Virology, Fields BN, Knipe DM. (eds). Raven Press: New York, 1990; 2079–2112. [Google Scholar]
  • 26. Payne LG, Kristenson K. Mechanism of vaccinia virus release and its specific inhibition by N1‐isonicotinoyl‐N2‐3‐methyl‐4‐chlorobenzoylhydrazine. J Virol 1979; 32: 614–622. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27. Blasco R, Moss B. Role of cell‐associated enveloped vaccinia virus in cell‐to‐cell spread. J Virol 1992; 66: 4170–4179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28. Patel DD, Pickup DJ, Joklik WK. Isolation of cowpox virus A‐type inclusions and characterization of their major protein component. Virology 1986; 149: 174–189. [DOI] [PubMed] [Google Scholar]
  • 29. Franke CA, Wilson EM, Hruby DE. Use of a cell‐free system to identify the vaccinia virus L1R gene product as the major late myristylated virion protein M25. J Virol 1990; 64: 5988–5996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30. Franke CA, Reynolds PL, Hruby DE. Fatty acid acylation of vaccinia virus proteins. J Virol 1989; 63: 4285–4291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31. Rosemond H, Moss B. Phosphoprotein component of vaccinia virions. J Virol 1973; 11: 961–970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32. Garon CF, Moss B. Glycoprotein synthesis in cells infected with vaccinia virus. II. A glycoprotein component of the virion. Virology 1971; 46: 233–246. [DOI] [PubMed] [Google Scholar]
  • 33. VanSlyke JK, Hruby DE. Posttranslational modification of vaccinia virus proteins. Curr Top Microbiol Immunol 1990; 163: 185–206. [DOI] [PubMed] [Google Scholar]
  • 34. Child SJ, Franke CA, Hruby DE. Inhibition of vaccinia virus replication by nicotinamide: evidence for ADP‐ribosylation of viral proteins. Virus Res 1988; 9: 119–132. [DOI] [PubMed] [Google Scholar]
  • 35. Stroobant P, Rice AP, Gullick WJ, Cheng DJ, Kerr IM, Waterfield MD. Purification and characterization of vaccinia virus growth factor. Cell 1985; 42: 383–393. [DOI] [PubMed] [Google Scholar]
  • 36. Chang W, Lim JG, Hellstrom I, Gentry LE. Characterization of vaccinia virus growth factor biosynthetic pathway with an antipeptide antiserum. J Virol 1988; 62: 1080–1083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37. Payne LG. Characterization of vaccinia virus glycoproteins by monoclonal antibody precipitation. Virology 1992; 187: 251–260. [DOI] [PubMed] [Google Scholar]
  • 38. Yang WP, Kao SY, Bauer WR. Biosynthesis and post‐translational cleavage of vaccinia virus structural protein VP8. Virology 1988; 167: 585–590. [PubMed] [Google Scholar]
  • 39. Holowczak JA, Joklik WK. Studies on the structural protein of vaccinia virus. Structural protein of virions and cores. Virology 1967; 33: 717–725. [DOI] [PubMed] [Google Scholar]
  • 40. Katz E, Moss B. Formation of a vaccinia virus structural polypeptide from a higher molecular weight precursor: inhibition by rifampicin. Proc Natl Acad Sci USA 1970; 66: 677–684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 41. Katz E, Moss B. Vaccinia virus structural polypeptide derived from a high‐molecular‐weight precursor: formation and integration into virus particles. J Virol 1970; 6: 717–726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 42. Moss B, Rosenblum EN. Letter: Protein cleavage and poxvirus morphogenesis: tryptic peptide analysis of core precursors accumulated by blocking assembly with rifampicin. J Mol Biol 1973; 81: 267–269. [DOI] [PubMed] [Google Scholar]
  • 43. Sarov I, Joklik WK. Studies on the nature and location of the capsid polypeptides of vaccinia virions. Virology 1972; 50: 579–592. [DOI] [PubMed] [Google Scholar]
  • 44. Pennington TH. Vaccinia virus polypeptide synthesis: sequential appearance and stability of pre‐ and post‐replicative polypeptides. J Gen Virol 1974; 25: 433–444. [DOI] [PubMed] [Google Scholar]
  • 45. Silver M, Dales S. Biogenesis of vaccina: interrelationship between post‐translational cleavage, virus assembly, and maturation. Virology 1982; 117: 341–356. [DOI] [PubMed] [Google Scholar]
  • 46. Rosel J, Moss B. Transcriptional and translational mapping and nucleotide sequence analysis of a vaccinia virus gene encoding the precursor of the major core polypeptide 4b. J Virol 1985; 56: 830–838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 47. Weir JP, Moss B. Regulation of expression and nucleotide sequence of a late vaccinia virus gene. J Virol 1984; 51: 662–669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 48. Goebel SJ, Johnson GP, Perkus MP, Davis SW, Winslow JP, Paoletti E. The complete DNA sequence of vaccinia virus. Virology 1990; 179: 247–66, 517–563. [DOI] [PubMed] [Google Scholar]
  • 49. VanSlyke JK, Franke CA, Hruby DE. Proteolytic maturation of vaccinia virus core proteins: identification of a conserved motif at the N termini of the 4b and 25K virion proteins. J Gen Virol 1991; 72: 411–426. [DOI] [PubMed] [Google Scholar]
  • 50. Binns M, Boursnell ME, Tomley FM, Campbell J. Analysis of the fowlpoxvirus gene encoding the 4b core polypeptide and demonstration that it possesses efficient promoter sequences. Virology 1989; 170: 288–291. [DOI] [PubMed] [Google Scholar]
  • 51. Vanslyke JK, Whitehead SS, Wilson EM, Hruby DE. The multistep proteolytic maturation pathway utilized by vaccinia virus P4a protein: a degenerate conserved cleavage motif within core proteins. Virology 1991; 183: 467–478. [DOI] [PubMed] [Google Scholar]
  • 52. Whitehead SS, Hruby DE. Differential utilization of a conserved motif for the proteolytic maturation of vaccinia virus proteins. Virology 1994; 200: 154–161. [DOI] [PubMed] [Google Scholar]
  • 53. Mercer J, Traktman P. Genetic and cell biological characterization of the vaccinia virus A30 and G7 phosphoproteins. J Virol 2005; 79: 7146–7161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 54. Vanslyke JK, Lee P, Wilson EM, Hruby DE. Isolation and analysis of vaccinia virus previrions. Virus Genes 1993; 7: 311–324. [DOI] [PubMed] [Google Scholar]
  • 55. Vanslyke JK, Hruby DE. Immunolocalization of vaccinia virus structural proteins during virion formation. Virology 1994; 198: 624–635. [DOI] [PubMed] [Google Scholar]
  • 56. Lee P, Hruby DE. Analysis of the role of the amino‐terminal peptide of vaccinia virus structural protein precursors during proteolytic processing. Virology 1995; 207: 229–233. [DOI] [PubMed] [Google Scholar]
  • 57. Fuerst TR, Niles EG, Studier FW, Moss B. Eukaryotic transient‐expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. Proc Natl Acad Sci USA 1986; 83: 8122–8126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 58. Prickett KS, Amberg DC, Hopp TP. A calcium‐dependent antibody for identification and purification of recombinant proteins. Biotechniques 1989; 7: 580–589. [PubMed] [Google Scholar]
  • 59. Lee P, Hruby DE. Proteolytic cleavage of vaccinia virus virion proteins. Mutational analysis of the specificity determinants. J Biol Chem 1994; 269: 8616–8622. [PubMed] [Google Scholar]
  • 60. Schechter I, Berger A. On the size of the active site in proteases. I. Papain. Biochem Biophys Res Commun 1967; 27: 157–162. [DOI] [PubMed] [Google Scholar]
  • 61. Whitehead SS, Hruby DE. A transcriptionally controlled trans‐processing assay: putative identification of a vaccinia virus‐encoded proteinase which cleaves precursor protein P25K. J Virol 1994; 68: 7603–7608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 62. Moss B. Vaccinia virus transcription In Transcription, Conway JARC. (ed.). Raven Press: New York, 1994; 185–206. [Google Scholar]
  • 63. Keck JG, Kovacs GR, Moss B. Overexpression, purification, and late transcription factor activity of the 17‐kilodalton protein encoded by the vaccinia virus A1L gene. J Virol 1993; 67: 5740–5748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 64. Keck JG, Baldick CJ, Moss B. Role of DNA replication in vaccinia virus gene expression: a naked template is required for transcription of three late trans‐ activator genes. Cell 1990; 61: 801–809. [DOI] [PubMed] [Google Scholar]
  • 65. Jones EV, Puckett C, Moss B. DNA‐dependent RNA polymerase subunits encoded within the vaccinia virus genome. J Virol 1987; 61: 1765–1771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 66. Kubo M, Imanaka T. Cloning and nucleotide sequence of the highly thermostable neutral protease gene from Bacillus stearothermophilus. J Gen Microbiol 1988; 134: 1883–1892. [DOI] [PubMed] [Google Scholar]
  • 67. Liu X, Yang J, Ghazi AM, Frey TK. Characterization of the zinc binding activity of the rubella virus nonstructural protease. J Virol 2000; 74: 5949–5956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 68. Tellinghuisen TL, Marcotrigiano J, Gorbalenya AE, Rice CM. The NS5A protein of hepatitis C virus is a zinc metalloprotein. J Biol Chem 2004; 279: 48576–48587. [DOI] [PubMed] [Google Scholar]
  • 69. Hedengren‐Olcott M, Byrd CM, Watson J, Hruby DE. The vaccinia virus G1L putative metalloproteinase is essential for viral replication in vivo. J Virol 2004; 78: 9947–9953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 70. Ansarah‐Sobrinho C, Moss B. Vaccinia virus G1 protein, a predicted metalloprotease, is essential for morphogenesis of infectious virions but not for cleavage of major core proteins. J Virol 2004; 78: 6855–6863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 71. Li SJ, Hochstrasser M. A new protease required for cell‐cycle progression in yeast. Nature 1999; 398: 246–251. [DOI] [PubMed] [Google Scholar]
  • 72. Condit RC, Motyczka A, Spizz G. Isolation, characterization, and physical mapping of temperature‐sensitive mutants of vaccinia virus. Virology 1983; 128: 429–443. [DOI] [PubMed] [Google Scholar]
  • 73. Kane EM, Shuman S. Vaccinia virus morphogenesis is blocked by a temperature‐sensitive mutation in the I7 gene that encodes a virion component. J Virol 1993; 67: 2689–2698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 74. Ericsson M, Cudmore S, Shuman S, Condit RC, Griffiths G, Locker JK. Characterization of ts 16, a temperature‐sensitive mutant of vaccinia virus. J Virol 1995; 69: 7072–7086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 75. Byrd CM, Bolken TC, Hruby DE. The vaccinia virus I7L gene product is the core protein proteinase. J Virol 2002; 76: 8973–8976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 76. Byrd CM, Bolken TC, Hruby DE. Molecular dissection of the vaccinia virus I7L core protein proteinase. J Virol 2003; 77: 11279–11283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 77. Byrd CM, Hruby DE. A conditional‐lethal vaccinia virus mutant demonstrates that the I7L gene product is required for virion morphogenesis. Virol J 2005; 2: 4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 78. Ansarah‐Sobrinho C, Moss B. Role of the I7 protein in proteolytic processing of vaccinia virus membrane and core components. J Virol 2004; 78: 6335–6343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 79. Byrd CM, Bolken TC, Mjalli AM, et al New class of orthopoxvirus antiviral drugs that block viral maturation. J Virol 2004; 78: 12147–12156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 80. Byrd CM, Hruby DE. Development of an in vitro cleavage assay system to examine vaccinia virus I7L cysteine proteinase activity. Virol J 2005; 2: 63. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Reviews in Medical Virology are provided here courtesy of Wiley

RESOURCES