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.

This article has been cited by other articles:

				X. Zhang, T. J. Green, J. Tsao, S. Qiu, and M. Luo Role of Intermolecular Interactions of Vesicular Stomatitis Virus Nucleoprotein in RNA Encapsidation J. Virol., January 15, 2008; 82(2): 674 - 682. [Abstract] [Full Text] [PDF]

				P. Walpita and C. J. Peters Cis-acting elements in the antigenomic promoter of Nipah virus J. Gen. Virol., September 1, 2007; 88(9): 2542 - 2551. [Abstract] [Full Text] [PDF]

				K. MacLellan, C. Loney, R. P. Yeo, and D. Bhella The 24-Angstrom Structure of Respiratory Syncytial Virus Nucleocapsid Protein-RNA Decameric Rings J. Virol., September 1, 2007; 81(17): 9519 - 9524. [Abstract] [Full Text] [PDF]

				M. Deng, J. N. Bragg, S. Ruzin, D. Schichnes, D. King, M. M. Goodin, and A. O. Jackson Role of the Sonchus Yellow Net Virus N Protein in Formation of Nuclear Viroplasms J. Virol., May 15, 2007; 81(10): 5362 - 5374. [Abstract] [Full Text] [PDF]

				A. A. V. Albertini, A. K. Wernimont, T. Muziol, R. B. G. Ravelli, C. R. Clapier, G. Schoehn, W. Weissenhorn, and R. W. H. Ruigrok Crystal Structure of the Rabies Virus Nucleoprotein-RNA Complex Science, July 21, 2006; 313(5785): 360 - 363. [Abstract] [Full Text] [PDF]

				C. Springfeld, G. Darai, and R. Cattaneo Characterization of the Tupaia Rhabdovirus Genome Reveals a Long Open Reading Frame Overlapping with P and a Novel Gene Encoding a Small Hydrophobic Protein J. Virol., June 1, 2005; 79(11): 6781 - 6790. [Abstract] [Full Text] [PDF]

				P. Kaukinen, V. Kumar, K. Tulimaki, P. Engelhardt, A. Vaheri, and A. Plyusnin Oligomerization of Hantavirus N Protein: C-Terminal {alpha}-Helices Interact To Form a Shared Hydrophobic Space J. Virol., December 15, 2004; 78(24): 13669 - 13677. [Abstract] [Full Text] [PDF]

				P. Liu, J. Yang, X. Wu, and Z. F. Fu Interactions amongst rabies virus nucleoprotein, phosphoprotein and genomic RNA in virus-infected and transfected cells J. Gen. Virol., December 1, 2004; 85(12): 3725 - 3734. [Abstract] [Full Text] [PDF]

				K. Lymperopoulos, C. Wirblich, I. Brierley, and P. Roy Sequence Specificity in the Interaction of Bluetongue Virus Non-structural Protein 2 (NS2) with Viral RNA J. Biol. Chem., August 22, 2003; 278(34): 31722 - 31730. [Abstract] [Full Text] [PDF]

				D. Escors, A. Izeta, C. Capiscol, and L. Enjuanes Transmissible Gastroenteritis Coronavirus Packaging Signal Is Located at the 5' End of the Virus Genome J. Virol., July 15, 2003; 77(14): 7890 - 7902. [Abstract] [Full Text] [PDF]

				X. Wu, X. Gong, H. D. Foley, M. J. Schnell, and Z. F. Fu Both Viral Transcription and Replication Are Reduced when the Rabies Virus Nucleoprotein Is Not Phosphorylated J. Virol., March 27, 2002; 76(9): 4153 - 4161. [Abstract] [Full Text] [PDF]

				X. Xu, W. Severson, N. Villegas, C. S. Schmaljohn, and C. B. Jonsson The RNA Binding Domain of the Hantaan Virus N Protein Maps to a Central, Conserved Region J. Virol., March 7, 2002; 76(7): 3301 - 3308. [Abstract] [Full Text] [PDF]

				Y. Jacob, E. Real, and N. Tordo Functional Interaction Map of Lyssavirus Phosphoprotein: Identification of the Minimal Transcription Domains J. Virol., October 15, 2001; 75(20): 9613 - 9622. [Abstract] [Full Text] [PDF]

				W. E. Severson, X. Xu, and C. B. Jonsson cis-Acting Signals in Encapsidation of Hantaan Virus S-Segment Viral Genomic RNA by Its N Protein J. Virol., March 15, 2001; 75(6): 2646 - 2652. [Abstract] [Full Text]

				G. Schoehn, F. Iseni, M. Mavrakis, D. Blondel, and R. W. H. Ruigrok Structure of Recombinant Rabies Virus Nucleoprotein-RNA Complex and Identification of the Phosphoprotein Binding site J. Virol., January 1, 2001; 75(1): 490 - 498. [Abstract] [Full Text]