Peptides that mimic the amino-terminal end of the rabies virus phosphoprotein have antiviral activity.
We wanted to develop a therapeutic approach against rabies disease by targeting the lyssavirus transcription/replication complex. Because this complex (nucleoprotein N-RNA template processed by the L polymerase and its cofactor, the phosphoprotein P) is similar to that of other negative-strand RNA viruses, we aimed to design broad-spectrum antiviral drugs that could be used as a complement to postexposure vaccination and immunotherapy. Recent progress in understanding the structure/function of the rabies virus P, N, and L proteins predicts that the amino-terminal end of P is an excellent target for destabilizing the replication complex because it interacts with both L (for positioning onto the N-RNA template) and N (for keeping N soluble, as needed for viral RNA encapsidation). Thus, peptides mimicking various lengths of the amino-terminal end of P have been evaluated, as follows: (i) for binding properties to the N-P-L partners by the two-hybrid method; (ii) for their capacity to inhibit the transcription/replication of a rabies virus minigenome encoding luciferase in BHK-21-T7 cells; and (iii) for their capacity to inhibit rabies virus infection of BHK-21-T7 cells and of two derivatives of the neuronal SK-N-SH cell line. Peptides P60 and P57 (the first 60 and first 57 NH2 residues of P, respectively) exhibited a rapid, strong, and long-lasting inhibitory potential on luciferase expression (>95% from 24 h to 55 h). P42 was less efficient in its inhibition level (75% for 18 to 30 h) and duration (40% after 48 h). The most promising peptides were synthesized in tandem with the Tat sequence, allowing cell penetration. Their inhibitory effects were observed on BHK-21-T7 cells infected with rabies virus and Lagos bat virus but not with vesicular stomatitis virus. In neuronal cells, a significant inhibition of both nucleocapsid inclusions and rabies virus release was observed.
Castel
Guillaume
G
Unité Postulante des Stratégies Antivirales, CNRS URA-3015, Institut Pasteur, 25 rue du Docteur Roux, 75724 Paris Cedex 15, France.
Chtéoui
Mohamed
M
Caignard
Grégory
G
Préhaud
Christophe
C
Méhouas
Stéphanie
S
Réal
Eléonore
E
Jallet
Corinne
C
Jacob
Yves
Y
Ruigrok
Rob W H
RW
Tordo
Noël
N
D000818:Animals; D000998:Antiviral Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D045744:Cell Line, Tumor; D006224:Cricetinae; D006801:Humans; D047468:Immunoprecipitation; D018832:Molecular Chaperones; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000821:virology; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D010750:Phosphoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011820:Rabies virus / Q000187:drug effects* / Q000472:pathogenicity; D020798:Two-Hybrid System Techniques; D015678:Viral Structural Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects*
D000998:Antiviral Agents; D018832:Molecular Chaperones; C103961:P phosphoprotein, Rabies virus; D010455:Peptides; D010750:Phosphoproteins; D015678:Viral Structural Proteins
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