title: Nuclear trafficking of macromolecules by an oligopeptide derived from Vpr of human immunodeficiency virus type-1.
abstract: Vpr, an accessory gene product of HIV-1, is incorporated into cells when added to the culture medium. Via such function Vpr has been shown to transduce a protein into cells that is expressed as a chimeric protein with Vpr. The domain required for protein transduction, however, remained to be clarified. Here we identified a sequence encompassing 52-78 amino acids of Vpr (C45D18) that enables nuclear trafficking of proteins. When chemically synthesized C45D18 was added to the culture medium of human cord blood mononuclear (CBMN) cells, most cells became positive for the incorporated C45D18. Furthermore, recombinant proteins conjugated with the C45D18 were efficiently transduced and transported to regions corresponding to the nucleus. Incorporation of C45D18-conjugated protein was observed within a few hours after addition of the protein, independent of cellular growth. Although it is well known that Tat-derived peptide has a transducing activity, C45D18 was more active than Tat peptide for trafficking proteins into cells. Taking together with results from FACS analysis revealing that more than 90% of CBMN cells were positive for X-gal staining after treatment of C45D18-conjugated beta-galactosidase, we propose that C45D18 translocates bioactive macromolecules directly into the nucleus.
authors.affiliation: Department of Intractable Diseases, International Medical Center of Japan, 1-21-1 Toyama, Shinjuku-ku, Tokyo 162-8655, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Taguchi
authors.forename: Mari
authors.initials: M
authors.lastname: Shimura
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Osawa
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Suzuki
authors.forename: Izuru
authors.initials: I
authors.lastname: Mizoguchi
authors.forename: Koitsu
authors.initials: K
authors.lastname: Niino
authors.forename: Fumimaro
authors.initials: F
authors.lastname: Takaku
authors.forename: Yukihito
authors.initials: Y
authors.lastname: Ishizaka
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000502:physiology*; D000595:Amino Acid Sequence; D016336:Gene Products, vpr / Q000737:chemistry* / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism*; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D015398:Signal Transduction / Q000502:physiology*; D013329:Structure-Activity Relationship
chemical_list: D016336:Gene Products, vpr; D008164:Luminescent Proteins; D046911:Macromolecular Substances; D049452:Green Fluorescent Proteins