The human cytomegalovirus glycoprotein B gene (ORF UL55) is expressed early in the infectious cycle Free

Abstract

Northern hybridizations were carried out using mRNA preparations of human cytomegalovirus (HCMV)- infected cultures and gene-specific antisense RNA probes for transcriptional analysis of the gene cluster composed of genes for DNA polymerase, glycoprotein B(gB), herpes simplex virus-infected cell protein 18.5 homologue p130 and a major DNA-binding protein corresponding to open reading frames (ORFs) UL54- UL57, respectively. Monocistronic transcripts of 5 kb and 3·7 kb were found for ORFs UL54 and UL55, respectively, and five additional high molecular mass overlapping transcripts of 14 kb, 10 kb, 10 kb, 8 kb and 6 kb were found. Mapping of 5′ ends showed that transcription was initiated at the expected distance downstream of predicted TATA elements; in the case of a UL56-specific transcript two potential initiation sites were identified. Transcription was found to terminate at the expected distance downstream of either of two prominent polyadenylation consensus motifs in the region of UL54. All transcripts were identified early in the infectious cycle, except for the UL55 (gB)-specific transcript of 3·7 kb which was not synthesized until late post-infection. However, specific immunoreactions demonstrated the presence of a gB-specific polypeptide early after infection in the absence of viral DNA synthesis. It is suggested that a bicistronic transcript of 8 kb encoded by ORFs UL55 and UL54 is involved in biosynthesis of early HCMV gB.

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1997-08-01
2024-03-29
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References

  1. Albrecht J. -C., Fleckenstein B. 1990; Structural organization of the conserved gene block of herpesvirus saimiri coding for DNA polymerase, glycoprotein B, and the major DNA binding protein. Virology 174:533–542
    [Google Scholar]
  2. Anders D., Gibson W. 1988; Location, transcript analysis, and partial nucleotide sequence of the cytomegalovirus gene encoding an early DNA-binding protein with similarities to ICP8 of herpes simplex virus type 1. Journal of Virology 62:1364–1372
    [Google Scholar]
  3. Baradaran K., Dabrowski C., Schaffer P. 1994; Transcriptional analysis of the region of the herpes simplex virus type 1 genome containing the UL8, UL9, and UL10 genes and identification of a novel delayed-early gene product, OBPC. Journal of Virology 68:4251–4261
    [Google Scholar]
  4. 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]
  5. Bogner E., Reschke M., Reis B., Mockenhaupt T., Radsak K. 1993; Identification of the gene product encoded by orf UL56 of the human cytomegalovirus genome. Virology 196:290–293
    [Google Scholar]
  6. Britt W., Vugler L. 1989; Processing of the gp55-116 envelope glycoprotein complex gB (g B) of human cytomegalovirus. Journal of Virology 58:403–410
    [Google Scholar]
  7. Cai W., Wu 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]
  8. Chang C., Malone C., Stinski M. 1989; A human cytomegalovirus early gene has three inducible promoters that are regulated differentially at various times after infection. Journal of Virology 63:281–290
    [Google Scholar]
  9. Chee M., Bankier A., Beck S., Bohni R., Brown C., Cerny R., Horsnell T., Hutchison C. III Kouzarides T., Martignetti E., Preddie E., Satchwell S., Tomlinson P., Weston K., Barrell B. 1990; Analysis of the protein coding content of the sequence of human cytomegalovirus AD169. Current Topics in Microbiology and Immunology 154:125–169
    [Google Scholar]
  10. 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]
  11. Engler-Blum G., Meier M., Frank J., Müller G. 1993; Reduction of background problems in nonradioactive Northern and Southern blot analyses enables higher sensitivity than 32P-based hybridizations. Analytical Biochemistry 210:235–244
    [Google Scholar]
  12. Fleckenstein B., Müller I., Collins J. 1982; Cloning of the complete human cytomegalovirus genome in cosmids. Gene 18:39–46
    [Google Scholar]
  13. Gong M., Kieff E. 1990; Intracellular trafficking of two major Epstein-Barr virus glycoproteins, gp350/320 and gp210. Journal of Virology 64:1507–1516
    [Google Scholar]
  14. Heilbronn R., Jahn G., Bürkle A., Freese U., Fleckenstein B., zurHausen H. 1987; Genomic localization, sequence analysis and transcription of the putative human cytomegalovirus DNA polymerase gene. Journal of Virology 61:119–124
    [Google Scholar]
  15. Herrold R., Marchini A., Fruehling S., Longnecker R. 1996; Glycoprotein 110, the EpsteinBarr virus homolog of herpes simplex virus glycoprotein B, is essential for Epstein-Barr virus replication in vivo. Journal of Virology 70:2049–2054
    [Google Scholar]
  16. Iwayama S., Yamamoto T., Furuya T., Kobayashi R., Ikuta K., Hirai K. 1994; Intracellular localization and DNA binding activity of a class of viral early phosphoproteins in human fibroblasts infected with human cytomegalovirus (Towne strain). Journal of General Virology 75:3309–3318
    [Google Scholar]
  17. Kemble G., McGormick A., Pereira L., Mocarski E. 1987; Acytomegalovirus protein with properties of herpes simplex virus ICP8: partial purification of the polypeptide and map position of the gene. Journal of Virology 61:3143–3153
    [Google Scholar]
  18. 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 tissue culture. Journal of Virology 65:5184–5189
    [Google Scholar]
  19. Kouzarides T., Bankier A., Satchwell S., Weston K., Tomlinson P., Barrell B. 1987a; Sequence and transcription analysis of the human cytomegalovirus DNA polymerase gene. Journal of Virology 61:125–133
    [Google Scholar]
  20. Kouzarides T., Bankier A., Satchwell S., Weston K., Tomlinson P., Barrell B. 1987b; Large -scale rearrangement of homologous regions in the genomes of HCMV and EBV. Virology 157:397–413
    [Google Scholar]
  21. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  22. Mach M., Utz U., Fleckenstein B. 1986; Mapping of the major glycoprotein gene of human cytomegalovirus. Journal of General Virology 67:1461–1467
    [Google Scholar]
  23. Mettenleiter T., Saalmüller A., Weiland F. 1993; Pseudorabies virus protein homologous to herpes simplex virus type 1 ICP 18.5 is necessary for capsid maturation. Journal of Virology 67:1236–1245
    [Google Scholar]
  24. Ohlin M., Sundquist V., Mach M., Wahren B., Borrebaeck C. 1993; Fine specificity of the human immune response to the major neutralizing epitopes expressed on cytomegalovirus gp58/116 (gB), as determined with human monoclonal antibodies. Journal of Virology 67:703–710
    [Google Scholar]
  25. Pederson N., Person S., Homa F. 1992; Analysis of the gB promoter of herpes simplex virus type 1 : high-level expression requires both an 89-base-pair promoter fragment and a nontranslated leader sequence. Journal of Virology 66:6226–6232
    [Google Scholar]
  26. Radsak K., Brücher K., Britt W., Shiou H., Schneider D., Kollert A. 1990; Nuclear compartmentation of glycoprotein B of human cytomegalovirus. Virology 177:515–522
    [Google Scholar]
  27. Reis B., Bogner E., Reschke M., Richter A., Mockenhaupt T., Radsak K. 1993; Stable constitutive expression of glycoprotein B (gpUL55) of human cytomegalovirus in permissive astrocytoma cells. Journal of General Virology 74:1371–1379
    [Google Scholar]
  28. Sambrook J., Fritsch E., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  29. Sanger F., Nicklen S., Coulson A. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, USA 74:5463–5467
    [Google Scholar]
  30. Schleiss M. 1994; Cloning and characterization of the guinea pig cytomegalovirus glycoprotein B gene. Virology 202:173–185
    [Google Scholar]
  31. Schleiss M. 1995; Sequence and transcriptional analysis of the guinea- pig cytomegalovirus DNA polymerase gene. Journal of General Virology 76:1827–1833
    [Google Scholar]
  32. 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]
  33. Tengelsen L., Pederson N., Shaver P., Wathen M., Homa F. 1993; Herpes simplex virus type 1 DNA cleavage and encapsidation require the product of the UL28 gene : isolation and characterization of two UL28 deletion mutants. Journal of Virology 67:3470–3480
    [Google Scholar]
  34. Vey M., Schäfer W., Reis B., Ohuchi R., Britt W., Garten W., Klenk H. -D., Radsak K. 1995; Proteolytic processing of human cytomegalovirus glycoprotein B (gpUL55) is mediated by the human endoprotease furin. Virology 206:746–749
    [Google Scholar]
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