1887

Abstract

Permanent cell lines showing homogeneous constitutive expression of glycoprotein B (gpUL55; gB) of human cytomegalovirus (HCMV) were selected, in the presence of geneticin, from human astrocytoma cells (U373) after transfection with recombinant pRC/CMV-gB carrying the complete coding sequence for HCMV gB and for aminoglycoside phosphotransferase. The biosynthesis and processing including specific proteolytic cleavage, formation of disulphide-linked oligomers as well as transport of recombinant gB in three of four established transformed cell lines essentially resembled that found in infected parental U373 except for eventual degradation after 2 h of gB synthesis. Analysis of the fourth transformant expressing uncleaved gB suggested that proteolytic cleavage is not required for normal intracellular transport. The stable transformants retained permissiveness for productive superinfection with HCMV. The application of cell lines transformed with mutagenized HCMV gB for the rescue of genetically engineered HCMV mutants is discussed.

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1993-07-01
2024-12-14
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References

  1. Blewett E, Misra V. 1991; Cleavage of the bovine herpesvirus glycoprotein B is not essential for its function. Journal of General Virology 72:2083–2090
    [Google Scholar]
  2. Bogner E, Reschke M, Reis B, Reis E, Britt W, Radsak K. 1992; Recognition of compartmentalized intracellular analogs of glycoprotein H of human cytomegalovirus. Archives of Virology 126:67–80
    [Google Scholar]
  3. Bonner WM, Laskey RA. 1974; A Film Detection Method for Tritium-Labelled Proteins and Nucleic Acids in Polyacrylamide Gels. European Journal of Biochemistry 46:83–88
    [Google Scholar]
  4. Britt WJ, Auger D. 1986; Synthesis and processing of the envelope gp55-116 complex of human cytomegalovirus. Journal of Virology 58:185–191
    [Google Scholar]
  5. Britt WJ, Vugler LG. 1989; 63:403–410 Processing of the gp55-116 envelope glycoprotein complex (gB) of human cytomegalovirus. Journal of Virology
    [Google Scholar]
  6. Britt WJ, Vugler L, Butfiloski EJ, Stephens EB. 1990; Cell surface expression of human cytomegalovirus (HCMV) gp55-116 (gB): use of HCMV-recombinant vaccinia virus-infected cells in analysis of the human neutralizing antibody response. Journal of Virology 63:403–410
    [Google Scholar]
  7. Cai WZ, Person S, Warner SC, Zhou JH, DeLuca NA. 1987; Linker-insertion nonsense and restriction-site deletion mutations of the gB glycoprotein gene of herpes simplex virus type 1. Journal of Virology 61:714–721
    [Google Scholar]
  8. Cai WH, Gu B, Person S. 1988; Role of glycoprotein B of herpes simplex virus type 1 in viral entry and cell fusion. Journal of Virology 62:2596–2604
    [Google Scholar]
  9. CampadelliFiume G, Arsenakis M, Farabegoli F, Roizman B. 1988a; Entry of herpes simplex virus 1 in BJ cells that constitutively express viral glycoprotein D is by endocytosis and results in degradation of the virus. Journal of Virology 62:159–167
    [Google Scholar]
  10. CampadelliFiume G, Avitabile E, Fini S, Stirpe D, Arsenakis M, Roizman E. 1988b; Herpes simplex virus glycoprotein D is sufficient to induce spontaneous pH-independent fusion in a cell line that constitutively expresses the glycoprotein. Virology 166:598–602
    [Google Scholar]
  11. CampadelliFiume G, Qi S, Avitabile E, Foà-Tomasi L, Brandimarti R, Roizman B. 1990; Glycoprotein D of herpes simplex virus encodes a domain which precludes penetration of cells expressing the glycoprotein by superinfecting herpes simplex virus. Journal of Virology 64:6070–6079
    [Google Scholar]
  12. Chen C, Okayama H. 1987; High-efficiency transformation of mammalian cells by plasmid DNA.. Molecular and Cellular Biology 7:2745–2752 1987
    [Google Scholar]
  13. Chomczynski P, Sacchi N. 1987; Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Analytical Biochemistry 162:156–159
    [Google Scholar]
  14. Cranage MP, Kouzarides T, Bankier AT, Satchwell S, Weston K, Tomlinson P, Barrell B, Hart H, Bell SE, Minson AC. 1986; Identification of the human cytomegalovirus glycoprotein B gene and induction of neutralizing antibodies via its expression in recombinant vaccinia virus.. The EMBO Journal 5:3057–3063
    [Google Scholar]
  15. Fleckenstein B, Müller I, Collins J. 1982; Cloning of the complete human cytomegalovirus genome in cosmids. Gene 18:39–46
    [Google Scholar]
  16. Garten W, Bosch F, Linder D, Rott R, Klenk HD. 1981; Proteolytic activation of the influenza virus hemagglutinin: The structure of the cleavage site and the enzymes involved in cleavage. Virology 115:361–374
    [Google Scholar]
  17. Gompels U, Minson A. 1989; Antigenic properties and cellular localization of herpes simplex virus glycoprotein H synthesized in a mammalian cell expression system. Journal of Virology 63:4744–4755
    [Google Scholar]
  18. Gretch D, Kari B, Rasmussen L, Gehrz R, Stinski M. 1988; Identification and characterization of three distinct families of glycoprotein complexes in the envelopes of human cytomegalovirus. Journal of Virology 62:875–881
    [Google Scholar]
  19. Hermiston T, Malone C, Stinski M. 1990; Human cytomegalovirus immediate-early two protein region involved in negative regulation of the major immediate-early promoter. Journal of Virology 64:3532–3536
    [Google Scholar]
  20. Johnson R, Spear P. 1989; Herpes simplex virus glycoprotein D mediates interference with herpes simplex virus infection. Journal of Virology 63:819–827
    [Google Scholar]
  21. Johnson R, Burke R, Gregory T. 1990; Soluble forms of herpes simplex virus glycoprotein D bind to a limited number of cell surface receptors and inhibit virus entry into cells. Journal of Virology 64:2569–2576
    [Google Scholar]
  22. Jones T, Muzithras V. 1992; A cluster of dispensable genes within the human cytomegalovirus genome short component: IRS1, US1 through US5, and the US6 family. Journal of Virology 66:2541–2546
    [Google Scholar]
  23. Jones T, Muzithras V, Gluzman Y. 1991; Replacement mutagenesis of the human cytomegalovirus genome: US10 and US11 gene products are nonessential. Journal of Virology 65:5860–5872
    [Google Scholar]
  24. Kari B, Radeke R, Gehrz R. 1992; Processing of human cytomegalovirus envelope glycoproteins in and egress of cytomegalovirus from human astrocytoma cells. Journal of General Virology 73:253–260
    [Google Scholar]
  25. Kollert-Jöns A, Bogner E, Radsak K. 1991; A 15-kilobase-pair region of the human cytomegalovirus genome which includes US1 through US13 is dispensable for growth in cell culture. Journal of Virology 65:5184–5189
    [Google Scholar]
  26. Laemmli UK. 1970; Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  27. Ligas M, Johnson D. 1988; A herpes simplex virus mutant in which glycoprotein D sequences are replaced by β-galactosidase sequences binds to but is unable to penetrate into cells. Journal of Virology 62:1486–1494
    [Google Scholar]
  28. Longnecker R, Roizman B. 1987; Clustering of genes dispensable for growth in culture in the S component of the HSV-1 genome. Science 236:573–576
    [Google Scholar]
  29. Mettenleiter T, Kern H, Rauh I. 1990; Isolation of a viable herpesvirus (pseudorabies virus) mutant specifically lacking all four known nonessential glycoproteins. Virology 179:498–503
    [Google Scholar]
  30. Peeters B, de Wind N, Broer R, Gielkens A, Moormann R. 1992; Glycoprotein H of pseudorabies virus is essential for entry and cell-to-cell spread of the virus. Journal of Virology 66:3888–3892
    [Google Scholar]
  31. Radsak K, Brücher KH, Britt W, Shiou H, Schneider D, Kollert A. 1990; Nuclear compartmentation of glycoprotein B of human cytomegalovirus. Virology 177:515–522
    [Google Scholar]
  32. Rapp F. 1983; The biology of cytomegaloviruses New York & London. The Herpesviruses vol 2 pp 1–66Edited by Roizman B. New York & London: Plenum Press;
    [Google Scholar]
  33. Rasmussen L., Nelson M., Neff M., Merigan T. 1988; Characterization of two different human cytomegalovirus glycoproteins which are targets for virus neutralizing antibody. Virology 163:308–318
    [Google Scholar]
  34. Rauh I, Mettenleiter T C. 1991; Pseudorabies virus glycoproteins gII and gp50 are essential for virus penetration. Journal of Virology 65:5348–5356
    [Google Scholar]
  35. Sambrook J, Fritsch E F, Maniatis T. 1989; 2nd edn. New York: Cold Spring Harbor Laboratory.. Molecular Cloning: A Laboratory Manual 65:5348–5356
    [Google Scholar]
  36. Sanger F., Nicklen S., Coulson A. R. 1977; 2nd edn. New York: Cold Spring Harbor Laboratory.. Molecular Cloning: A Laboratory Manual 65:5348–5356
    [Google Scholar]
  37. Spaete R. R., Mocarski E. S. 1987; Insertion and deletion mutagenesis of the human cytomegalovirus genome.. Proceedings of the National Academy of Sciences 84:7213–7217
    [Google Scholar]
  38. Spaete R., Thayer R., Probert W., Masiarz F., Chamberlain S., Rasmussen L., Merigan T., Pachl C. 1988; Human cytomegalovirus strain towne glycoprotein B is processed by proteolytic cleavage. Virology 167:207–225
    [Google Scholar]
  39. Spaete R., Saxena A., Scott P., Song G.,, Probert W.,, Britt W.,, Gibson W.,, Rasmussen L., Pachl C. 1990; Sequence requirements for proteolytic processing of glycoprotein B of human cytomegalovirus strain Towne. Journal of Virology 64:2922–2931
    [Google Scholar]
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