A Tat-grafted anti-nucleic acid antibody acquires nuclear-localization property and a preference for TAR RNA |
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Authors: | Jeong Jong-Geun Kim Dong-Sik Kim Yong-Sung Kwon Myung-Hee |
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Institution: | aDepartment of Microbiology, Ajou University School of Medicine, San 5, Woncheon-dong, Yeongtong-gu, Suwon 443-749, South Korea;bDepartment of Molecular Science and Technology, Ajou University, San 5, Woncheon-dong, Yeongtong-gu, Suwon 443-749, South Korea |
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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 (H3Tat-3D8) by replacing complementarity-determining region 3 (CDR3) within the VH domain of the 3D8 scFv with a Tat48–60 peptide (GRKKRRQRRRPPQ). H3Tat-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. |
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Keywords: | Anti-nucleic acid antibody Nucleic acid-hydrolyzing antibody 3D8 scFv Tat peptide Tat-grafted antibody |
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