A symmetry mismatch at the site of RNA packaging in the polymerase complex of dsRNA bacteriophage phi6.
The polymerase complex of the enveloped double-stranded RNA (dsRNA) bacteriophage phi6 fulfils a similar function to those of other dsRNA viruses such as Reoviridae. The phi6 complex comprises protein P1, which forms the shell, and proteins P2, P4 and P7, which are involved in RNA synthesis and packaging. Icosahedral reconstructions from cryo-electron micrographs of recombinant polymerase particles revealed a clear dodecahedral shell and weaker satellites. Difference imaging demonstrated that these weak satellites were the sites of P4 and P2 within the complex. The structure determined by icosahedral reconstruction was used as an initial model in an iterative reconstruction technique to examine the departures from icosahedral symmetry. This approach showed that P4 and P2 contribute to structures at the 5-fold positions of the icosahedral P1 shell which lack 5-fold symmetry and appear in variable orientations. Reconstruction of isolated recombinant P4 showed that it was a hexamer with a size and shape matching the satellite. Symmetry mismatch between the satellites and the shell could play a role in RNA packaging akin to that of the portal vertex of dsDNA phages in DNA packaging. This is the first example of dsRNA virus in which the structure of the polymerase complex has been determined without the assumption of icosahedral symmetry. Our result with phi6 illustrates the symmetry mismatch which may occur at the sites of RNA packaging in other dsRNA viruses such as members of the Reoviridae.
de Haas
F
F
The Structural Biology Programme, European Molecular Biology Laboratory, Meyerhofstrasse 1, Heidelberg, 69117, Germany.
Paatero
A O
AO
Mindich
L
L
Bamford
D H
DH
Fuller
S D
SD
D017126:Bacteriophage phi 6 / Q000235:genetics* / Q000648:ultrastructure; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry*; D007091:Image Processing, Computer-Assisted; D008854:Microscopy, Electron / Q000379:methods; D012330:RNA, Double-Stranded / Q000737:chemistry*; D012367:RNA, Viral / Q000737:chemistry*; D015203:Reproducibility of Results; D014764:Viral Proteins / Q000737:chemistry / Q000302:isolation & purification
D012330:RNA, Double-Stranded; D012367:RNA, Viral; D014764:Viral Proteins; C080701:protein P4 (Pseudomonas); D012321:DNA-Directed RNA Polymerases