Ultrastructural and functional analyses of recombinant influenza virus ribonucleoproteins suggest dimerization of nucleoprotein during virus amplification.
Influenza virus ribonucleoproteins (RNPs) were reconstituted in vivo from cloned cDNAs expressing the three polymerase subunits, the nucleoprotein (NP), and short template RNAs. The structure of purified RNPs was studied by electron microscopy and image processing. Circular and elliptic structures were obtained in which the NP and the polymerase complex could be defined. Comparison of the structure of RNPs of various lengths indicated that each NP monomer interacts with approximately 24 nucleotides. The analysis of the amplification of RNPs with different lengths showed that those with the highest replication efficiency contained an even number of NP monomers, suggesting that the NP is incorporated as dimers into newly synthesized RNPs.
Ortega
J
J
Centro Nacional de Biotecnología (CSIC), Campus de Cantoblanco, 28049 Madrid, Spain.
Martín-Benito
J
J
Zürcher
T
T
Valpuesta
J M
JM
Carrascosa
J L
JL
Ortín
J
J
D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D003001:Cloning, Molecular; D017931:DNA Primers; D019281:Dimerization; D008854:Microscopy, Electron; D009975:Orthomyxoviridae / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism / Q000648:ultrastructure; D012261:Ribonucleoproteins / Q000302:isolation & purification / Q000502:physiology* / Q000648:ultrastructure
D017931:DNA Primers; D011994:Recombinant Proteins; D012261:Ribonucleoproteins