Molecular cloning, complete sequence, and biological characterization of a xenotropic murine leukemia virus constitutively released from the human B-lymphoblastoid cell line DG-75

• Published in: Virology (New York, N.Y.) Vol. 308; no. 1; pp. 83 - 91
• Main Authors: Raisch, Kevin P, Pizzato, Massimo, Sun, Hai-Yuan, Takeuchi, Yasuhiro, Cashdollar, L.William, Grossberg, Sidney E
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 30.03.2003
• Subjects: 3T3 Cells; Animals; B-Lymphocytes; Cell Line; Cell Line, Transformed; Cloning, Molecular; Coculture Techniques; Complete genome; DG-75 MLV; Evi-2; Genes, Viral; Genome, Viral; Humans; Leukemia Virus, Murine - genetics; Leukemia Virus, Murine - physiology; Lymphoblastoid; Mice; MLV; Molecular Sequence Data; Primer-binding site; Receptors, G-Protein-Coupled; Receptors, Virus - analysis; Retrovirus; RNA, Messenger - analysis; RNA, Viral - genetics; Sequence Homology, Nucleic Acid; Viral Interference; Viral Structural Proteins - genetics; Virus Replication; DG-75 MLV; Retrovirus; MLV; Lymphoblastoid; Complete genome; Primer-binding site; Evi-2
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1016/S0042-6822(02)00074-0

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.