Selection and characterization of human immunodeficiency virus type 1 mutants that are resistant to inhibition by the transdominant negative RevM10 protein.
Intracellular immunization with RevM10, a transdominant negative form of the Rev protein, efficiently inhibits human immunodeficiency virus (HIV) replication in vitro and gene therapy protocols that use this modality are currently being evaluated in human clinical trials. Development of resistance to this kind of therapy has not been previously reported. Here we show that RevM10-resistant HIV type 1 (HIV-1) variants can be selected by in vitro passage of HIV-1 in a T-lymphoblastoid cell line constitutively expressing RevM10. Unexpectedly, the selected variants showed changes in the Rev response element (RRE) but no changes in Rev. Replacement of the wild-type RRE with a mutated RRE resulted in a virus that showed increased resistance to RevM10. After repeated passages of the resistant variant in cells expressing RevM10, a virus with an additional mutation in the viral vpu gene was selected. Surprisingly, a virus containing only this vpu mutation also showed some resistance to inhibition by RevM10.
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Myles H. Thaler Center for AIDS and Human Retrovirus Research and the Department of Microbiology, University of Virginia, Charlottesville, Virginia 22908, USA.
Rekosh
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Hammarskjöld
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002460:Cell Line; D004279:DNA, Viral; D015694:Gene Products, rev / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter; D014644:Genetic Variation; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D054298:Human Immunodeficiency Virus Proteins; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D011533:Proviruses / Q000235:genetics; D012367:RNA, Viral; D020218:Response Elements; D054334:Viral Regulatory and Accessory Proteins / Q000235:genetics; D054321:rev Gene Products, Human Immunodeficiency Virus; D000086082:Viroporin Proteins
D004279:DNA, Viral; D015694:Gene Products, rev; D054298:Human Immunodeficiency Virus Proteins; D012367:RNA, Viral; D054334:Viral Regulatory and Accessory Proteins; D054321:rev Gene Products, Human Immunodeficiency Virus; C514859:vpu protein, Human immunodeficiency virus 1; D000086082:Viroporin Proteins