Equine infectious anemia virus and human immunodeficiency virus DNA synthesis in vitro: characterization of the endogenous reverse transcriptase reaction.
The endogenous reverse transcriptase reaction of equine infectious anemia virus (EIAV) has been studied, and conditions allowing synthesis of full-length minus-strand DNA have been determined. In contrast to results reported for other retroviruses, synthesis of EIAV full-length minus-strand DNA was not impaired by high concentrations of Nonidet P-40, a nonionic detergent used to make the virion envelope permeable. All components of the reaction were titrated for maximum synthesis of complete minus strands, and a time course under the standardized conditions was determined. Minor subgenomic bands were observed in some cases, and both the size and proportion varied with reaction conditions. Conditions established for full-length EIAV DNA synthesis also allowed full-genome-length human immunodeficiency virus type 1 DNA synthesis. The human immunodeficiency virus type 1 DNA product contained a greater proportion of reverse transcripts that were shorter than the complete virus genome. Also in contrast to EIAV, the endogenous synthesis of high-molecular-weight human immunodeficiency virus type 1 DNA was drastically reduced at Nonidet P-40 concentrations above 0.02%. These results indicated that a detergent-stable core is not a property shared by all lentiviruses. The EIAV virion synthetic machinery is unusually stable and provides a convenient system for further in vitro study of reverse transcription.
Borroto-Esoda
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Division of Virology, Wellcome Research Laboratories, Burroughs Wellcome Co., Research Triangle Park, North Carolina 27709.
Boone
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LR
D000085:Acetates / Q000378:metabolism; D019342:Acetic Acid; D000818:Animals; D001483:Base Sequence; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D004261:DNA Replication / Q000187:drug effects; D004279:DNA, Viral / Q000096:biosynthesis*; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D006736:Horses; D066298:In Vitro Techniques; D004860:Infectious Anemia Virus, Equine / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D008969:Molecular Sequence Data; D009711:Nucleotides / Q000378:metabolism; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D014771:Virion / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D014779:Virus Replication / Q000187:drug effects
D000085:Acetates; D004279:DNA, Viral; D009711:Nucleotides; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40; D012194:RNA-Directed DNA Polymerase; D019342:Acetic Acid