Expression and cellular localization of the Nef protein from human immunodeficiency virus-1 in stably transfected B-cells.
Nef protein, encoded by the regulatory nef gene of human immunodeficiency virus type 1 (HIV-1), was expressed in the B-cell line Raji. The cells were stably transfected with plasmids containing the nef transcriptional cassette. They expressed Nef with an Mr of 27,000; the yield could be augmented by incubation with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate. The intracellular localization of Nef was analyzed applying immunofluorescence microscopy using a confocal laser scanning microscope. The antigen was stained with a monoclonal antibody directed against the N-terminal part of Nef. The experiments revealed that in non-dividing cells Nef is present both in the cytoplasm and the nucleus while in dividing cells the viral protein is present in the cytoplasm and at the nuclear membrane.
Kienzle
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Abteilung Virologie, Universität Homburg, Saar, Federal Republic of Germany.
Bachmann
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M
Müller
W E
WE
Müller-Lantzsch
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N
D001402:B-Lymphocytes / Q000737:chemistry* / Q000187:drug effects; D005455:Fluorescent Antibody Technique; D015693:Gene Products, nef / Q000096:biosynthesis* / Q000187:drug effects / Q000302:isolation & purification; D005822:Genetic Vectors / Q000235:genetics; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008853:Microscopy / Q000379:methods; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured; D054311:nef Gene Products, Human Immunodeficiency Virus
D015693:Gene Products, nef; D054311:nef Gene Products, Human Immunodeficiency Virus; D013755:Tetradecanoylphorbol Acetate