The anti-HIV pentameric pseudopeptide HB-19 binds the C-terminal end of nucleolin and prevents anchorage of virus particles in the plasma membrane of target cells.
The multivalent pseudopeptide HB-19 that binds the cell-surface-expressed nucleolin is a potent inhibitor of human immunodeficiency virus (HIV) infection by blocking virus particle attachment and thus anchorage in the plasma membrane. We show that cross-linking of surface-bound HB-19A (like HB-19 but with a modified template) results in aggregation of HB-19A with surface nucleolin. Consistent with its specific action, HB-19A binding to different types of cells reaches saturation at concentrations that have been reported to result in inhibition of HIV infection. By using Chinese hamster ovary mutant cell lines, we confirm that the binding of HB-19A to surface nucleolin is independent of heparan and chondroitin sulfate proteoglycans. In vitro generated full-length nucleolin was found to bind HB-19A, whereas the N-terminal part containing the acidic amino acid stretches of nucleolin did not. The use of various deletion constructs of the C-terminal part of nucleolin then permitted the identification of the extreme C-terminal end of nucleolin, containing repeats of the amino acid motif, RGG, as the domain that binds HB-19A. Finally, a synthetic peptide corresponding to the last C-terminal 63 amino acids was able to inhibit HIV infection at the stage of HIV attachment to cells, thus suggesting that this domain could be functional in the HIV anchorage process.
Nisole
Sébastien
S
Unité de Virologie et Immunologie Cellulaire (URA 1930 CNRS), Plateau Technique, Institut Pasteur, 28 Rue du Dr. Roux, 75724 Paris Cedex 15, France.
Said
Elias A
EA
Mische
Claudia
C
Prevost
Marie-Christine
MC
Krust
Bernard
B
Bouvet
Philippe
P
Bianco
Alberto
A
Briand
Jean-Paul
JP
Hovanessian
Ara G
AG
D000595:Amino Acid Sequence; D000818:Animals; D019380:Anti-HIV Agents / Q000378:metabolism* / Q000494:pharmacology; D001483:Base Sequence; D016466:CHO Cells; D002462:Cell Membrane / Q000187:drug effects; D006224:Cricetinae; D017931:DNA Primers; D015497:HIV-1 / Q000187:drug effects* / Q000502:physiology; D008561:Membrane Fusion / Q000187:drug effects*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010455:Peptides; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism* / Q000494:pharmacology; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D014771:Virion / Q000187:drug effects / Q000502:physiology; D000097796:Nucleolin
D019380:Anti-HIV Agents; D017931:DNA Primers; D010455:Peptides; D010750:Phosphoproteins; D011506:Proteins; D016601:RNA-Binding Proteins; D000097796:Nucleolin; C121944:HB-19 peptide