1887

Abstract

Here we examine by polymerase chain reaction amplification followed by cloning and sequence analyses selected regions of the human herpesvirus 6 (HHV-6) genome which contain human telomeric repeats (TTAGGG). We determine the relative number, arrangement and orientation of the repeats in the unit length genome, in concatemeric replicative intermediates and in heterogeneous (het) regions. We also examine distribution of the repeats in the entire genome (159 kb) and their orientation relative to DNA packaging motifs and the origin of lytic replication. In the prototype orientation the HHV-6 repeat is the related complement, TAACCC. We show that tandem array sof this repeat are present in the right and left long direct repeats (DR and DR, 8 kb each) which bound the long unique sequence (U, 143 kb). Within each DR there is a left terminal imperfect tandem array and a right terminal perfect tandem array (58 copies). In DR they are each adjacent to DNA packaging motifs, pac1 and pac2, described for herpes simplex virus and human cytomegalovirus, in the arrangement pac1-imperfect repeat-7.2 kb-perfect repeat-pac2. Five independent clones were isolated and sequence determined from junctions of concatemeric replicative intermediates which showed adjacent pac2 and pac1 motifs surrounded by telomeric repeats. Favoured cleavage sites for unit length genomes were indicated which avoided cleavage within the repeats. Analyses of the complete genome showed no tandem repeats within U but did show a polar distribution of monomeric copies and related sequences around the origin of replication, with an effect on the overall base composition. The implications for virus replication are discussed.

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1995-02-01
2021-10-23
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References

  1. Bankier A. T., Weston K. M., Barrell B. G. 1988; Random cloning and sequencing by the M13/ dideoxynucleotide chain termination method. Methods in Enzvmology 155:51–93
    [Google Scholar]
  2. Blackburn E. H. 1991; Structure and function of telomeres. Nature 350:560–573
    [Google Scholar]
  3. Brown W. R. A., MacKinnon P. J., Villasante A., Spurr N., Buckle V. J., Dobson M. J. 1990; Structure and polymorphism of human telomere-associated DNA. Cell 63:119–132
    [Google Scholar]
  4. Carrigan D. R., Know K. K., Russler M. A. 1990; Suppression of human immunodeficiency virus type 1 replication by human herpesvirus-6. Journal of Infectious Diseases 162:844–851
    [Google Scholar]
  5. Carrigan D. R., Drobyski W. R., Russler S. K., Tapper M. A., Knox K. K., Ash R. C. 1991; Interstitial pneumonitis associated with human herpesvirus-6 infection after marrow transplantation. Lancet 338:147–149
    [Google Scholar]
  6. Cone R. W., Hackman R. C., Huang M. L. W., Bowden R. A., Myers J. D., Metcalp M., Zeh J., Ashley R., Corey L. 1993; Human herpesvirus 6 in lung tissue from patients with pneumonitis after bone marrow transplantation. New England Journal of Medicine 329:156–161
    [Google Scholar]
  7. Deiss L. P., Chou J., Frenkel N. 1986; Functional domains within the a sequence involved in the cleavage-packaging of herpes simplex virus DNA. Journal of Virology 59:605–618
    [Google Scholar]
  8. Downing R. G., Sewankambo N., Serwadda D., Honess R. W., Crawford D., Jarrett R., Griffen B. E. 1987; Isolation of human lymphotropic herpesviruses from Uganda. Lancet ii:390
    [Google Scholar]
  9. Drobyski W. R., Dunne W. M., Burd E. M., Knox K. K., Ash R. C., Horowitz M. M., Flemenbery N., Carrigan D. R. 1993; Human herpesvirus-6 (HHV-6) infection in allogeneic bone marrow transplant recipients. I. Evidence of a marrow suppressive role for HHV-6 in vivo . Journal of Infectious Diseases 167:735–739
    [Google Scholar]
  10. Efstathiou S., Lawrence G. L., Brown C. M., Barrell B. G. 1992; Identification of homologues to the human cytomegalovirus US22 gene family in human herpesvirus 6. Journal of General Virology 73:1661–1671
    [Google Scholar]
  11. Ensoli B., Lusso P., Schachter F., Josephs S. F., Rappaport J., Negro F., Gallo R. C., Wong-Staal F. 1989; Human herpesvirus-6 increases HIV-1 expression in co-infected T cells via nuclear factors binding to the HIV-1 enhancer. EMBO Journal 8:3019–3027
    [Google Scholar]
  12. Gleeson T. J., Staden R. 1991; An x-window and UNIX implementation of our sequence analysis package. Computing Applied Biosciences 7:398
    [Google Scholar]
  13. Gompels U. A., Carss A. L., Sun N., Arrand J. R. 1992; Infectivity determinants encoded in a conserved gene block of human herpesvirus-6. Journal of DNA Sequencing and Mapping 3:25–39
    [Google Scholar]
  14. Gompels U. A., Carrigan D. C., Carss A. L., Arno J. 1993; Two groups of human herpesvirus 6 identified by sequence analyses of laboratory strains and variants from Hodgkin’s lymphoma and bone marrow transplant patients. Journal of General Virology 74:613–622
    [Google Scholar]
  15. Gompels U. A., Luxton J., Knox K. K., Carrigan D. R. 1994a; Chronic bone marrow suppression in immunocompetent adult by human herpesvirus 6. Lancet 343:735–736
    [Google Scholar]
  16. Gompels U. A., Nicholas J., Lawrence G., Thomson B., Jones M., Efstathiou S. 1994b; (Manuscript in preparation.).
    [Google Scholar]
  17. Greider C. W. 1993; Telomeres and telomerase in small eukaryotes. Symposium of the Society for General Microbiology 50:31–42
    [Google Scholar]
  18. Gui J.-F., Lane W. S., Fu X.-D. 1994; A serine kinase regulates intracellular localization of splicing factors in the cell cycle. Nature 369:678–682
    [Google Scholar]
  19. Harrington L. A., Greider C. W. 1991; Telomerase primer specificity and chromosome healing. Nature 353:451–454
    [Google Scholar]
  20. Kemble G. W., Mocarski E. 1989; A host cell protein binds to a highly conserved sequence element (pac-2) within the cytomegalovirus a sequence. Journal of Virology 63:4715–4728
    [Google Scholar]
  21. Kishi M., Harada H., Takahashi M., Tanaka A., Hayashi M., Nonoyama M., Josephs S. F., Buchbinder A., Schacter F., Ablashi D. V., Wong-Staal F., Salahuddin Z., Gallo R. C. 1988; A repeat sequence, GGGTTA, is shared by DNA of human herpesvirus 6 and Marek’s disease virus. Journal of Virology 62:4824–4827
    [Google Scholar]
  22. Kozak M. 1984; Compilation and analyses of sequences upstream from the translational start site of eukaryotic mRNAs. Nucleic Acids Research 12:857–872
    [Google Scholar]
  23. Lawrence G. L., Chee L. M., Craxton M. A., Gompels U. A., Honess R. W., Barrell B. G. 1990; Human herpesvirus 6 is closely related to human cytomegalovirus. Journal of Virology 64:287–299
    [Google Scholar]
  24. Lawrence G. L. 1991; Sequence analysis of the human herpesvirus-6 genome. PhD thesis University of Cambridge;
    [Google Scholar]
  25. Lusso P., Ensoli B., Markham P. D., Ablashi D. V., Salahuddin Z., Tschachler E., Wong-Staal F., Gallo R. C. 1989; Productive dual infection of human CD4+ T lymphocytes by HIV-1 and HIV-6. Nature 337:370–373
    [Google Scholar]
  26. Martin M. E. D., Thomson B. J., Honess R. W., Craxton M. A., Gompels U. A., Liu M.-Y., Littler E., Arrand J. R., Teo I., Jones M. D. 1991; The genome of human herpesvirus 6: maps of unit-length and concatemeric genomes for nine restriction endonucleases. Journal of General Virology 72:157–168
    [Google Scholar]
  27. Messing J. 1983; New M13 vectors for cloning. Methods in Enzymology 101:20–78
    [Google Scholar]
  28. Neipel F., Ellinger K., Fleckenstein B. 1991; The unique region of the human herpesvirus 6 genome is essentially collinear with the UL segment of human cytomegalovirus. Journal of General Virology 72:2293–2297
    [Google Scholar]
  29. Nicholas J., Martin M. E. D. 1994; Nucleotide sequence analysis of a 38-5-kilobase-pair region of the genome of human herpesvirus 6 encoding human cytomegalovirus immediate-early gene homologs and transactivating functions. Journal of Virology 68:597–610
    [Google Scholar]
  30. Okuno T., Takahashi K., Balachandra K., Shiraki K., Yamanishi K., Takahashi M., Baba K. 1989; Seroepidemiology of human herpesvirus 6 infection in normal children and adults. Journal of Clinical Microbiology 27:651–653
    [Google Scholar]
  31. Pearson W. R., Lipman D. J. 1988; Improved tools for biological sequence comparison. Proceedings of the National Academy of Sciences, U.S.A 85:2444–2448
    [Google Scholar]
  32. Salahuddin S. Z., Ablashi C. V., Markaham P. D., Josephs S. F., Sturzenegger S., Kaplan M., Halligan G., Biberfield P., Wong-Staal F., Kramarsky B., Gallo R. C. 1986; Isolation of a new virus, HBLV, in patients with lymphoproliferative disorders. Science 234:596–601
    [Google Scholar]
  33. Salazar M., Champoux J. J., Reid B. R. 1993; Sugar conformations at hybrid duplex junctions in HIV-1 and Okazaki fragments. Biochemistry 32:739–744
    [Google Scholar]
  34. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain terminating inhibitors. Proceedings of the National Academy of Sciences, U.S.A 74:5463–5467
    [Google Scholar]
  35. Takahashi K., Sonoda S., Higashi K., Kondo T., Takahashi H., Takashashi M., Yamanishi K. 1989; Predominant CD4 T-lymphocyte tropism of human herpesvirus-6 related virus. Journal of Virology 63:3161–3163
    [Google Scholar]
  36. Tedder T. S., Briggs M., Cameron C. H., Honess R. W., Robertson D., Whittle H. 1987; A novel lymphotropic herpesvirus. Lancet ii:390–391
    [Google Scholar]
  37. Thomson B., Dewhurst S., Gray D. 1993; Accession no. L22334-6..
    [Google Scholar]
  38. Thomson B. J., Dewhurst S. D., Gray D. 1994; Structure and heterogeneity of the a sequences of human herpesvirus 6 strain variants U1102 and Z29 and identification of human telomeric repeat sequences at the genomic termini. Journal of Virology 68:3007–3014
    [Google Scholar]
  39. Varmuza S. L., Smiley J. R. 1985; Signals for site-specific cleavage of herpes simplex virus DNA: maturation involves two separate cleavage events at sites distal to the recognition site. Cell 41:792–802
    [Google Scholar]
  40. Weber B., Collins C., Robbins C., Magenis R. E., Delaney A. D., Gray J. W., Hayden M. R. 1990; Characterization and organization of DNA sequences adjacent to the human telomere associated repeat (TTAGGG)n. Nucleic Acids Research 18:3353–3361
    [Google Scholar]
  41. Yamanishi K., Okuno T., Shiraki K., Takahashi M., Kondo T., Asano Y., Kurata T. 1988; Identification of human herpesvirus-6 as the causal agent for exanthem subitum. Lancet i1065–1067
    [Google Scholar]
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