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 (DRL and DRR, 8 kb each) which bound the long unique sequence (UL, 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 UL 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.
BankierA. T.,
WestonK. M.,
BarrellB. G.1988; Random cloning and sequencing by the M13/ dideoxynucleotide chain termination method. Methods in Enzvmology 155:51–93
BrownW. R. A.,
MacKinnonP. J.,
VillasanteA.,
SpurrN.,
BuckleV. J.,
DobsonM. J.1990; Structure and polymorphism of human telomere-associated DNA. Cell 63:119–132
CarriganD. R.,
KnowK. K.,
RusslerM. A.1990; Suppression of human immunodeficiency virus type 1 replication by human herpesvirus-6. Journal of Infectious Diseases 162:844–851
CarriganD. R.,
DrobyskiW. R.,
RusslerS. K.,
TapperM. A.,
KnoxK. K.,
AshR. C.1991; Interstitial pneumonitis associated with human herpesvirus-6 infection after marrow transplantation. Lancet 338:147–149
ConeR. W.,
HackmanR. C.,
HuangM. L. W.,
BowdenR. A.,
MyersJ. D.,
MetcalpM.,
ZehJ.,
AshleyR.,
CoreyL.1993; Human herpesvirus 6 in lung tissue from patients with pneumonitis after bone marrow transplantation. New England Journal of Medicine 329:156–161
DeissL. P.,
ChouJ.,
FrenkelN.1986; Functional domains within the a sequence involved in the cleavage-packaging of herpes simplex virus DNA. Journal of Virology 59:605–618
DowningR. G.,
SewankamboN.,
SerwaddaD.,
HonessR. W.,
CrawfordD.,
JarrettR.,
GriffenB. E.1987; Isolation of human lymphotropic herpesviruses from Uganda. Lancet ii:390
DrobyskiW. R.,
DunneW. M.,
BurdE. M.,
KnoxK. K.,
AshR. C.,
HorowitzM. M.,
FlemenberyN.,
CarriganD. 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
EfstathiouS.,
LawrenceG. L.,
BrownC. M.,
BarrellB. G.1992; Identification of homologues to the human cytomegalovirus US22 gene family in human herpesvirus 6. Journal of General Virology 73:1661–1671
EnsoliB.,
LussoP.,
SchachterF.,
JosephsS. F.,
RappaportJ.,
NegroF.,
GalloR. C.,
Wong-StaalF.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
GompelsU. A.,
CarssA. L.,
SunN.,
ArrandJ. R.1992; Infectivity determinants encoded in a conserved gene block of human herpesvirus-6. Journal of DNA Sequencing and Mapping 3:25–39
GompelsU. A.,
CarriganD. C.,
CarssA. L.,
ArnoJ.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
GompelsU. A.,
LuxtonJ.,
KnoxK. K.,
CarriganD. R.1994a; Chronic bone marrow suppression in immunocompetent adult by human herpesvirus 6. Lancet 343:735–736
KembleG. W.,
MocarskiE.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
KishiM.,
HaradaH.,
TakahashiM.,
TanakaA.,
HayashiM.,
NonoyamaM.,
JosephsS. F.,
BuchbinderA.,
SchacterF.,
AblashiD. V.,
Wong-StaalF.,
SalahuddinZ.,
GalloR. 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
LawrenceG. L.,
CheeL. M.,
CraxtonM. A.,
GompelsU. A.,
HonessR. W.,
BarrellB. G.1990; Human herpesvirus 6 is closely related to human cytomegalovirus. Journal of Virology 64:287–299
LussoP.,
EnsoliB.,
MarkhamP. D.,
AblashiD. V.,
SalahuddinZ.,
TschachlerE.,
Wong-StaalF.,
GalloR. C.1989; Productive dual infection of human CD4+ T lymphocytes by HIV-1 and HIV-6. Nature 337:370–373
MartinM. E. D.,
ThomsonB. J.,
HonessR. W.,
CraxtonM. A.,
GompelsU. A.,
LiuM.-Y.,
LittlerE.,
ArrandJ. R.,
TeoI.,
JonesM. 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
NeipelF.,
EllingerK.,
FleckensteinB.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
NicholasJ.,
MartinM. 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
OkunoT.,
TakahashiK.,
BalachandraK.,
ShirakiK.,
YamanishiK.,
TakahashiM.,
BabaK.1989; Seroepidemiology of human herpesvirus 6 infection in normal children and adults. Journal of Clinical Microbiology 27:651–653
SalahuddinS. Z.,
AblashiC. V.,
MarkahamP. D.,
JosephsS. F.,
SturzeneggerS.,
KaplanM.,
HalliganG.,
BiberfieldP.,
Wong-StaalF.,
KramarskyB.,
GalloR. C.1986; Isolation of a new virus, HBLV, in patients with lymphoproliferative disorders. Science 234:596–601
SangerF.,
NicklenS.,
CoulsonA. R.1977; DNA sequencing with chain terminating inhibitors. Proceedings of the National Academy of Sciences, U.S.A 74:5463–5467
TakahashiK.,
SonodaS.,
HigashiK.,
KondoT.,
TakahashiH.,
TakashashiM.,
YamanishiK.1989; Predominant CD4 T-lymphocyte tropism of human herpesvirus-6 related virus. Journal of Virology 63:3161–3163
ThomsonB. J.,
DewhurstS. D.,
GrayD.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
VarmuzaS. L.,
SmileyJ. 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
WeberB.,
CollinsC.,
RobbinsC.,
MagenisR. E.,
DelaneyA. D.,
GrayJ. W.,
HaydenM. R.1990; Characterization and organization of DNA sequences adjacent to the human telomere associated repeat (TTAGGG)n. Nucleic Acids Research 18:3353–3361
YamanishiK.,
OkunoT.,
ShirakiK.,
TakahashiM.,
KondoT.,
AsanoY.,
KurataT.1988; Identification of human herpesvirus-6 as the causal agent for exanthem subitum. Lanceti1065–1067