Expression of human polyomavirus JC T antigen by an adenovirus hybrid vector and its binding to DNA sequences encompassing the JC virus origin of DNA replication
In the search for factors that influence the outcome of human polyomavirus JC (JCV) infection, the roles not only of host-related immunological control but also of virus-dependent regulatory steps have to be taken into account. Besides cell-specific control of early expression of the multifunctional virus protein large tumour antigen (T Ag), control mechanisms involve individual steps of the DNA replication process. For the analysis of T Ag DNA binding, the protein was expressed by an adenovirus hybrid vector in the 293 cell line to provide saturating amounts of JCV T Ag. After determination of the size and immunoreactivity, functional activity was analysed by specific DNA binding. To avoid the interference of cellular proteins, T Ag was immunoprecipitated prior to the reaction. Binding to T Ag-binding sites I and II within a 141 bp DNA segment in the control region was analysed using deletion mutants of a JCV subtype from brain tissue of a patient with fatal central nervous system disease. The specificity of the binding was confirmed by recombinant T Ag binding to origin of DNA replication (ori) sequences of wild-type JCV genomes. These data document that recombinant T Ag overexpressed by the adenovirus vector in eukaryotic cells was JCV-specific, had the expected length and exhibited specific ori-binding activity, thus providing the essential tool for future analysis of virus-host interactions at the level of viral DNA replication.
AmirhaeriS.,
WohlrabF.,
MajorE. O.,
WellsR.1988; Unusual DNA structure in the regulatory region of the human papovirus JC virus. Journal of Virology 62:922–931
BorowiecJ. A.,
DeanF. B.,
HurwitzJ.1991; Differential induction of structural changes in the simian virus 40 origin of replication by T antigen. Journal of Virology 65:1228–1235
FeigenbaumL.,
KhaliliK.,
MajorE.,
KhouryG.1987; Regulation of the host range of human papovavirus JCV. Proceedings of the National Academy of SciencesU.S.A. 84:3695–3698
FiersW.,
ContrerasR.,
FIaegemanG.,
RogiersR.,
Van de VoordeA.,
Van HeuverswynH.,
Van HerrewegheJ.,
VolckaertG.,
YsebaertM.1978; Complete nucleotide sequence of SV40 DNA. Science293113–120
FrisqueR. J.,
WhiteF. A. III 1992; The molecular biology of JC virus, causative agent of progressive multifocal leukoencephalopathy. In Molecular Neurovirology25–158RoosR. P.
Clifton, New Jersey: The Humana Press;
GrahamF. L.,
SmileyJ.,
RussellW. C.,
NairnR.1977; Characteristics of a human cell line transformed by DNA from human adenovirus type 5. Journal of General Virology 36:59–72
KerrD. A.,
ChangC. -F.,
GordonJ.,
BjornstiM. -A.,
KhaliliK.1993; Inhibition of human neurotropic virus (JCV) DNA replication in glial cells by camptothecin. Virology 196:612–618
LamarcheN.,
MassieB.,
RicherM.,
ParadisH.,
LangelierY.1990; High level expression in 293 cells of the herpes simplex virus type 2 ribonucleotide reductase subunit 2 using an adenovirus vector. Journal of General Virology 71:1785–1795
LiJ. J.,
KellyT. J.1985; Simian virus 40 DNA replication in vitro: specificity of initiation and evidence for bidirectional rep¬lication. Molecular and Cellular Biology 6:3086–3093
McVeyD.,
BrizuelaL.,
MohrI.,
MarshakD. R.,
GluzmanY.,
BeachD.1989; Phosphorylation of large tumour antigen by cdc2 stimulates SV40 DNA replication. Nature 341:503–507
MajorE. O.,
AmemiyaK.,
TornatoreC. S.,
HouffS. A.,
BergerJ. R.1992; Pathogenesis and molecular biology of progressive multifocal leukoencephalopathy, the JC virus-induced demyelinating disease of the human brain. Clinical Microbiology Reviews 5:49–73
MartinJ. D.,
KingD. M.,
SlauchJ. M.,
FrisqueR. J.1985; Differences in regulatory sequences of naturally occurring JC virus variants. Journal of Virology 53:306–311
MyersC.,
FrisqueR. J.,
ArthurR. R.1989; Direct isolation and characterization of JC virus from urine samples of renal and bone marrow patients. Journal of Virology 63:4445–1449
ParsonsR.,
AndersonM. E.,
TegtmeyerP.1990; Three domains in the simian virus 40 core origin orchestrate the binding, melting, and DNA helicase activities of T antigen. Journal of Virology 64:509–518
PreciousB.,
RussellW. C.1985; Growth, purification and titration of adenovirus. In Virology, a Practical Approach193–205MahyB. W. J.
Oxford: IRL Press;
RigbyP. W. J.,
LaneD. P.1983; Structure and function of simian virus 40 large T antigen. In Advances in Viral Oncology331–57KleinG.
New York: Raven Press;
SimmonsD. T.,
LoeberG.,
TegtmeyerP.1990; Four major sequence elements of simian virus 40 large T antigen coordinate its specific and non-specific DNA binding. Journal of Virology 64:1973–1983
SimmonsD. T.,
Wun-KimK.,
YoungW.1990; Identification of simian virus 40 T antigen residues important for specific and non- specific binding to DNA and for helicase activity. Journal of Virology 64:4858–1865
SockE.,
WegnerM.,
FortunatoE. A.,
GrummtF.1993; Large T-antigen and sequences within the regulatory region of JC virus both contribute to the features of JC virus DNA replication. Virology 197:537–548
StonerG. L.,
RyschkewitschC. F.,
WalkerD. L.,
SofferD.,
WebsterH. F.1988; A monoclonal antibody to SV40 large T antigen labels a nuclear antigen in JC virus-transformed cells and in progressive multifocal leucoencephalopathy (PML) brain infected with JC virus. Journal of Neuroimmunology 17:331–345
TaborS.,
RichardsonC. C.1987; DNA sequence analysis with a modified T7 DNA polymerase. Proceedings of the National Academy of SciencesU.S.A. 84:4767–4771
WalkerD. L.,
FrisqueR. J.1986; The biology and molecular biology of JC virus. In The Papovaviridae: The Polyomaviruses327–377SalzmanN. P.
New York: Plenum Press;
WalkerD. L.,
PadgettB. L.,
ZuRheinG. M.,
AlbertA. D.,
MarshR. F.1973; Human polyomavirus (JC): induction of brain tumors in hamsters. Science 181:674–676
YogoY.,
KitamuraT.,
SugimotoC.,
UekiT.,
AsoY.,
HaraK.,
TaguchiF. C.1990; Isolation of a possible archetypal JC virus DNA sequence from nonimmunocompromised individuals. Journal of Virology 64:3139–3143
Expression of human polyomavirus JC T antigen by an adenovirus hybrid vector and its binding to DNA sequences encompassing the JC virus origin of DNA replication