Antibody fragments selected by phage display against the nuclear localization signal of the HIV-1 Vpr protein inhibit nuclear import in permeabilized and intact cultured cells.
The HIV-1 Vpr protein harbors a nuclear localization signal in its N-terminal domain. A peptide bearing this domain and which is designated VprN has been used as a target to screen a phage display single chain Fv (scFv) library. Here we report the isolation of anti-VprN scFv fragments from this library. The purified scFv fragments were able to bind the VprN peptide in an ELISA-based system and to inhibit VprN-mediated nuclear import in permeabilized as well as in intact microinjected cells. Furthermore, the anti-VprN scFv fragments recognized the full-length recombinant Vpr protein and inhibited its nuclear import. The same scFv fragments did not inhibit nuclear import mediated by the nuclear localization signal of the SV40 large T-antigen demonstrating a specific effect. The use of the described inhibitory anti-VprN scFv fragments to study nuclear import of viral karyophilic proteins and their therapeutic potential is discussed.
Krichevsky
A
A
Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel.
Graessmann
A
A
Nissim
A
A
Piller
S C
SC
Zakai
N
N
Loyter
A
A
D021581:Active Transport, Cell Nucleus; D000818:Animals; D022801:Complementarity Determining Regions; D016336:Gene Products, vpr / Q000276:immunology* / Q000502:physiology; D015483:HIV Antibodies / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology; D019913:Nuclear Localization Signals / Q000276:immunology*; D019151:Peptide Library; D014407:Tumor Cells, Cultured; D054325:vpr Gene Products, Human Immunodeficiency Virus
D022801:Complementarity Determining Regions; D016336:Gene Products, vpr; D015483:HIV Antibodies; D007128:Immunoglobulin Fragments; D019913:Nuclear Localization Signals; D019151:Peptide Library; C055966:immunoglobulin Fv; D054325:vpr Gene Products, Human Immunodeficiency Virus