title: Isolation and characterisation of the rabies virus N degrees-P complex produced in insect cells.
abstract: When the nucleoprotein (N) of nonsegmented negative-strand RNA viruses is expressed in insect cells, it binds to cellular RNA and forms N-RNA complexes just like viral nucleocapsids. However, in virus-infected cells, N is prevented from binding to cellular RNA because a soluble complex is formed between N and the viral phosphoprotein (P), the N degrees -P complex. N is only released from this complex for binding to newly made viral or complementary RNA. We coexpressed rabies virus N and P proteins in insect cells and purified the N degrees -P complex. Characterisation by gel filtration, polyacrylamide gel electrophoresis, analytical ultracentrifugation, native mass spectroscopy, and electron microscopy showed that the complex consists of one N protein plus two P proteins, i.e., an N degrees -P(2) complex.
authors.affiliation: EMBL Grenoble Outstation, B.P. 181, 38042 Grenoble Cedex 9, France.
authors.forename: Manos
authors.initials: M
authors.lastname: Mavrakis
authors.forename: Frédéric
authors.initials: F
authors.lastname: Iseni
authors.forename: Catherine
authors.initials: C
authors.lastname: Mazza
authors.forename: Guy
authors.initials: G
authors.lastname: Schoehn
authors.forename: Christine
authors.initials: C
authors.lastname: Ebel
authors.forename: Marc
authors.initials: M
authors.lastname: Gentzel
authors.forename: Thomas
authors.initials: T
authors.lastname: Franz
authors.forename: Rob W H
authors.initials: RW
authors.lastname: Ruigrok
mesh_terms: D000818:Animals; D002850:Chromatography, Gel; D008854:Microscopy, Electron; D008970:Molecular Weight; D019251:Nucleocapsid / Q000737:chemistry / Q000302:isolation & purification* / Q000502:physiology; D019590:Nucleocapsid Proteins; D010750:Phosphoproteins / Q000737:chemistry / Q000302:isolation & purification* / Q000502:physiology; D011994:Recombinant Proteins / Q000302:isolation & purification; D018411:Spodoptera; D014764:Viral Proteins / Q000302:isolation & purification*
chemical_list: D019590:Nucleocapsid Proteins; D010750:Phosphoproteins; D011994:Recombinant Proteins; D014764:Viral Proteins; C099601:nucleocapsid protein, Rabies virus