A Tat-grafted anti-nucleic acid antibody acquires nuclear-localization property and a preference for TAR RNA

Journal Article · Fri Mar 18 04:00:00 EDT 2011 · Biochemical and Biophysical Research Communications

DOI:https://doi.org/10.1016/J.BBRC.2011.02.054· OSTI ID:22204834

Jeong, Jong-Geun ^[1]; Kim, Dong-Sik; Kim, Yong-Sung ^[2]; Kwon, Myung-Hee ^[1]

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1. Department of Microbiology, Ajou University School of Medicine, San 5, Woncheon-dong, Yeongtong-gu, Suwon 443-749 (Korea, Republic of)
2. Department of Molecular Science and Technology, Ajou University, San 5, Woncheon-dong, Yeongtong-gu, Suwon 443-749 (Korea, Republic of)

Highlights: {yields} We generate '{sub H3}Tat-3D8' by grafting Tat{sub 48-60} peptide to VH CDR of 3D8 scFv antibody. {yields} {sub H3}Tat-3D8 antibody retains nucleic acid binding and hydrolyzing activities. {yields} {sub H3}Tat-3D8 acquires a preference for TAR RNA structure. {yields} Properties of Tat{sub 48-60} is transferred to an antibody via Tat-grafting into a CDR. -- 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 ({sub H3}Tat-3D8) by replacing complementarity-determining region 3 (CDR3) within the VH domain of the 3D8 scFv with a Tat{sub 48-60} peptide (GRKKRRQRRRPPQ). {sub 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.

• OSTI ID: 22204834
• Journal Information: Biochemical and Biophysical Research Communications, Journal Name: Biochemical and Biophysical Research Communications Journal Issue: 3 Vol. 406; ISSN 0006-291X; ISSN BBRCA9
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
AIDS VIRUS
ANTIBODIES
CELL NUCLEI
DNA
GRAFTS
HELA CELLS
PEPTIDES
RNA