Molecular cloning, complete sequence, and biological characterization of a xenotropic murine leukemia virus constitutively released from the human B-lymphoblastoid cell line DG-75.
A previously undetected retrovirus has been isolated from the human Epstein-Barr virus (EBV)-negative, B-lymphoblastoid DG-75 cell line, widely used for EBV gene transfection studies. The complete 8207-base genome of the DG-75 retrovirus was molecularly cloned from viral mRNA and sequenced (Accession No. AF221065). Northern blot analysis with probes specific for the putative RU-5, gag, pol, and env regions identified a full-length viral RNA and spliced env mRNA. DG-75 viral RNA was isolated from the DG-75 cell sublines UW and KAR, but not from the HAD subline. The DG-75 retrovirus was isolated with primer-binding sites that match tRNA(Thr) and tRNA(Gln2). Homology searches revealed homology to (i) xenotropic NZB-9-1 env mRNA, (ii) Moloney-MLV pol region, and (iii) a truncated Evi-2 endogenous proviral sequence gag and pol region. Viral interference and infectivity assays confirmed the xenotropic nature of the DG-75 retrovirus. The DG-75 retrovirus is the first isolate of an exogenous xenotropic MLV in which the full-length genomic sequence has been characterized.
Raisch
Kevin P
KP
Department of Microbiology and Molecular Genetics, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA.
Pizzato
Massimo
M
Sun
Hai Yuan
HY
Takeuchi
Yasuhiro
Y
Cashdollar
L William
LW
Grossberg
Sidney E
SE
D016475:3T3 Cells; D000818:Animals; D001402:B-Lymphocytes; D002460:Cell Line; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D018920:Coculture Techniques; D005814:Genes, Viral; D016679:Genome, Viral; D006801:Humans; D009052:Leukemia Virus, Murine / Q000235:genetics* / Q000502:physiology; D051379:Mice; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D012367:RNA, Viral / Q000235:genetics; D043562:Receptors, G-Protein-Coupled; D011991:Receptors, Virus / Q000032:analysis; D012689:Sequence Homology, Nucleic Acid; D014762:Viral Interference; D015678:Viral Structural Proteins / Q000235:genetics*; D014779:Virus Replication; D000089842:Xenotropic and Polytropic Retrovirus Receptor
D012333:RNA, Messenger; D012367:RNA, Viral; D043562:Receptors, G-Protein-Coupled; D011991:Receptors, Virus; D015678:Viral Structural Proteins; D000089842:Xenotropic and Polytropic Retrovirus Receptor