Recombinant HIV-1 nucleocapsid protein p15 produced as a fusion protein with glutathione S-transferase in Escherichia coli mediates dimerization and enhances reverse transcription of retroviral RNA.
Human immunodeficiency virus 1 (HIV-1) nucleocapsid protein p15 was produced as a fusion protein with glutathione S-transferase (GST) in Escherichia coli. Rapid purification of GST::p15 in an active form by one-step glutathione-agarose chromatography was accomplished in the presence of an antioxidant. Recombinant p15 fused to GST was shown to stimulate the dimerization of viral RNA. HIV-1 reverse transcriptase-catalyzed in vitro synthesis of minus-strand cDNA from synthetic human tRNA(Lys3UUU) and natural bovine tRNA(Lys3SUU) primer molecules was enhanced by GST::p15. GST produced in E.coli revealed no effect with respect to RNA dimerization and cDNA synthesis, demonstrating that both activities reside in the p15 portion of the fusion protein.
D000595:Amino Acid Sequence; D001483:Base Sequence; D002213:Capsid / Q000737:chemistry / Q000378:metabolism*; D003001:Cloning, Molecular; D004247:DNA / Q000096:biosynthesis; D004268:DNA-Binding Proteins / Q000378:metabolism; D015683:Gene Products, gag / Q000737:chemistry / Q000378:metabolism*; D005982:Glutathione Transferase / Q000235:genetics; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000235:genetics*; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins; D012357:RNA, Transfer, Lys / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins / Q000737:chemistry / Q000378:metabolism*; D016335:Zinc Fingers; D054301:gag Gene Products, Human Immunodeficiency Virus
D004268:DNA-Binding Proteins; D015683:Gene Products, gag; D019590:Nucleocapsid Proteins; D012357:RNA, Transfer, Lys; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; C060295:p15 gag protein, Human immunodeficiency virus 1; D004247:DNA; D005982:Glutathione Transferase; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase