title: A Tat-grafted anti-nucleic acid antibody acquires nuclear-localization property and a preference for TAR RNA.
abstract: The 3D8 single chain variable fragment (3D8 scFv) is an anti-nucleic acid antibody that can hydrolyze nucleic acids and enter the cytosol of cells without reaching the nucleus. The Tat peptide, derived from the basic region of the HIV-1 Tat protein, translocates to cell nuclei and has TAR RNA binding activity. In this study, we generated a Tat-grafted antibody ((H₃)Tat-3D8) by replacing complementarity-determining region 3 (CDR3) within the VH domain of the 3D8 scFv with a Tat₄₈₋₆₀ peptide (GRKKRRQRRRPPQ). (H₃)Tat-3D8 retained the DNA-binding and DNA-hydrolyzing activity of the scFv, and translocated to the nuclei of HeLa cells and preferentially recognized TAR RNA. Thus, the properties associated with the Tat peptide were transferred to the antibody via Tat-grafting without loss of the intrinsic DNA-binding and hydrolyzing activities of the 3D8 scFv antibody.
authors.affiliation: Department of Microbiology, Ajou University School of Medicine, Suwon 443-749, South Korea.
authors.forename: Jong-Geun
authors.initials: JG
authors.lastname: Jeong
authors.forename: Dong-Sik
authors.initials: DS
authors.lastname: Kim
authors.forename: Yong-Sung
authors.initials: YS
authors.lastname: Kim
authors.forename: Myung-Hee
authors.initials: MH
authors.lastname: Kwon
mesh_terms: D000974:Antibodies, Antinuclear / Q000276:immunology* / Q000378:metabolism; D001483:Base Sequence; D002467:Cell Nucleus / Q000276:immunology* / Q000378:metabolism; D004247:DNA / Q000276:immunology; D015497:HIV-1; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D012367:RNA, Viral / Q000276:immunology* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000276:immunology* / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000276:immunology / Q000378:metabolism; D000099253:Trans-Activation Responsive RNA-Binding Protein
chemical_list: D000974:Antibodies, Antinuclear; D004247:DNA; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D057127:Single-Chain Antibodies; D000099253:Trans-Activation Responsive RNA-Binding Protein