Mapping the Peste des Petits Ruminants virus nucleoprotein: Identification of two domains involved in protein self-association

Bodjo Sanne Charles, Lelenta Mamadou, Couacy-Hymann Emmanuel, Kwiatek Olivier, Albina Emmanuel, Gargani Daniel, Libeau Geneviève, Diallo Adama. 2008. Mapping the Peste des Petits Ruminants virus nucleoprotein: Identification of two domains involved in protein self-association. Virus Research, 131 (1) : 23-32.

https://doi.org/10.1016/j.virusres.2007.08.005

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Résumé : For Mononegavirales, the template for transcription and replication is not the naked RNA but the nucleoprotein (N) encapsidated genomic and anti-genomic RNA. Because of this central role in the replication of these viruses, N has been the subject of numerous structural and functional mapping studies. Here, we report on the cloning of the Peste des Petits Ruminants virus (PPRV) N gene into the baculovirus vector and the expression of the protein in insect cells. By electron microscopy observation, we have shown that this recombinant PPRV N forms nucleocapsid-like particles in insect cells in the absence of other PPRV proteins, as reported for other paramyxoviruses. As it is known that the formation of these particles is first linked to the self-assembly of N, we have made several deletions in the PPRV N gene and expressed these mutants in insect cells. Analysis of these proteins by immunoprecipitation and electron microscopy observation enabled us to map the N¿N interaction domains into two regions of PPRV N: aa 1¿120 and 146¿241. The fragment aa 121¿145, which is not conserved within the morbillivirus group, is also required for the formation/stability of the nucleocapsid helical structure.

Mots-clés Agrovoc : peste des petits ruminants, virus peste petits ruminants, nucléoprotéine, morbillivirus, identification, carte génétique, interaction génique, Clonage moléculaire, expression des gènes

Classification Agris : L73 - Maladies des animaux
L10 - Génétique et amélioration des animaux

Champ stratégique Cirad : Axe 4 (2005-2013) - Santé animale et maladies émergentes

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