Importin-alpha promotes passage through the nuclear pore complex of human immunodeficiency virus type 1 Vpr.
Viral protein R (Vpr) of human immunodeficiency virus type 1 has potent karyophilic properties, but details of the mechanism by which it enters the nucleus remain to be clarified. We reported previously that two regions, located between residues 17 and 34 (alphaH1) and between residues 46 and 74 (alphaH2), are indispensable for the nuclear localization of Vpr. Here, we reveal that a chimeric protein composed of the nuclear localization signal of Vpr, glutathione S-transferase, and green fluorescent protein was localized at the nuclear envelope and then entered the nucleus upon addition of importin-alpha. An in vitro transport assay using a series of derivatives of importin-alpha demonstrated that the carboxyl terminus was required for this nuclear import process. We also showed that Vpr interacts with importin-alpha through alphaH1 and alphaH2; only the interaction via alphaH1 is indispensable for the nuclear entry of Vpr. These observations indicate that importin-alpha functions as a mediator for the nuclear entry of Vpr.
Kamata
Masakazu
M
Retrovirus Research Unit, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan. aida@riken.jp.
Nitahara-Kasahara
Yuko
Y
Miyamoto
Yoichi
Y
Yoneda
Yoshihiro
Y
Aida
Yoko
Y
D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D016336:Gene Products, vpr / Q000378:metabolism*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D015497:HIV-1 / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D022022:Nuclear Pore / Q000378:metabolism*; D025941:Protein Interaction Mapping; D021382:Protein Sorting Signals; D021381:Protein Transport; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship; D028901:alpha Karyopherins / Q000502:physiology*; D054325:vpr Gene Products, Human Immunodeficiency Virus
D016336:Gene Products, vpr; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D028901:alpha Karyopherins; D054325:vpr Gene Products, Human Immunodeficiency Virus; D049452:Green Fluorescent Proteins
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