Isolation and characterisation of the rabies virus N degrees-P complex produced in insect cells.
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.
Mavrakis
Manos
M
EMBL Grenoble Outstation, B.P. 181, 38042 Grenoble Cedex 9, France.
Iseni
Frédéric
F
Mazza
Catherine
C
Schoehn
Guy
G
Ebel
Christine
C
Gentzel
Marc
M
Franz
Thomas
T
Ruigrok
Rob W H
RW
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*
D019590:Nucleocapsid Proteins; D010750:Phosphoproteins; D011994:Recombinant Proteins; D014764:Viral Proteins; C099601:nucleocapsid protein, Rabies virus