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Structure of the nucleoprotein binding domain of Mokola virus phosphoprotein.

DOI: 10.1128/JVI.01520-09 DOI Help
PMID: 19906936 PMID Help

Authors: René Assenberg (Wellcome Trust Centre for Human Genetics, University of Oxford, U.K.) , O Delmas (WHO Collaborating Centre for Reference and Research on Rabies, Institut Pasteur, Paris, France) , Jingshan Ren (Wellcome Trust Centre for Human Genetics, University of Oxford, U.K) , Pierre-olivier Vidalain , Anil Verma (Wellcome Trust Centre for Human Genetics, University of Oxford, U.K) , Florence Larrous , Stephen C Graham (Wellcome Trust Centre for Human Genetics, University of Oxford, U.K.) , Frédéric Tangy , Jonathan Grimes (Division of Structural Biology, University of Oxford) , Hervé Bourhy (WHO Collaborating Centre for Reference and Research on Rabies, Institut Pasteur, Paris, France)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of Virology , VOL 84 (2) , PAGES 1089-1096

State: Published (Approved)
Published: December 2010

Abstract: Mokola virus (MOKV) is a nonsegmented, negative-sense RNA virus that belongs to the Lyssavirus genus and Rhabdoviridae family. MOKV phosphoprotein P is an essential component of the replication and transcription complex and acts as a cofactor for the viral RNA-dependent RNA polymerase. P recruits the viral polymerase to the nucleoprotein-bound viral RNA (N-RNA) via an interaction between its C-terminal domain and the N-RNA complex. Here we present a structure for this domain of MOKV P, obtained by expression of full-length P in Escherichia coli, which was subsequently truncated during crystallization. The structure has a high degree of homology with P of rabies virus, another member of Lyssavirus genus, and to a lesser degree with P of vesicular stomatitis virus (VSV), a member of the related Vesiculovirus genus. In addition, analysis of the crystal packing of this domain reveals a potential binding site for the nucleoprotein N. Using both site-directed mutagenesis and yeast two-hybrid experiments to measure P-N interaction, we have determined the relative roles of key amino acids involved in this interaction to map the region of P that binds N. This analysis also reveals a structural relationship between the N-RNA binding domain of the P proteins of the Rhabdoviridae and the Paramyxoviridae.

Journal Keywords: Crystallization ; Crystallography ; X-Ray ; Escherichia; Lyssavirus ; Models ; Molecular ; Mutagenesis ; Site-Directed ; Nucleocapsid; Phosphoproteins ; Structure-Activity; Two-Hybrid; Viral Proteins

Subject Areas: Biology and Bio-materials, Medicine


Instruments: I03-Macromolecular Crystallography

Added On: 04/04/2012 12:08

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