Journal of General Virology, Vol 79, 1005-1013, Copyright © 1998 by Society for General Microbiology

ARTICLES

Identification of a region of the rabies virus N protein involved in direct binding to the viral RNA

A Kouznetzoff, M Buckle and N Tordo
Laboratoire des Lyssavirus, Institut Pasteur, Paris, France.

In rabies virus, the ribonucleoprotein complex (RNP), the RNA genome (- ) and the antigenome (+) are specifically coated by the viral nucleoprotein (N protein), forming the template for transcription and replication bythe viral RNA polymerase. This specific encapsidation starts at the 5' ends of the RNAs. To investigate domains of the N protein that govern binding specificity, we tested in vitro the ability of both full-length and truncated forms of the N protein to interact with a synthetic RNA probe corresponding to the 5' end of the antigenome. UV-LASER cross-linking, which covalently links RNA and proteins in intimate contact, showed that the entire N protein (450 aa) and the NH2-terminal 376 aa (t42) contained all of the determinants for specific interaction. It was demonstrated by affinity chromatography that a peptide near the COOH terminus of t42 (position 298352), which is located in the most conserved region of Rhabdoviridae N proteins, bound directly to the viral RNA. However, no significant sequence similarity was detected between this peptide and known RNA binding proteins in the databases. This suggests both that N proteins may possess a new type of RNA binding motif and that protein folding contributes to the architecture of the RNA binding site.

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