Roles of the minor capsid protein P7 in the assembly and replication of double-stranded RNA bacteriophage phi6.
The polymerase complexes of double-stranded RNA (dsRNA) viruses are multifunctional RNA processing machineries that carry out viral genome packaging, replication, and transcription. The polymerase complex forms the innermost virion shell and is structurally related in dsRNA viruses infecting a diversity of host organisms. In this study, we analyzed the properties and functions of the minor polymerase complex protein P7 of dsRNA bacteriophage phi6 using terminally truncated P7 polypeptides and an in vitro self-assembly system established for the phi6 polymerase complex. The N-terminally truncated P7 failed to dimerize, whereas C-terminally truncated P7 polypeptides formed functional dimers that were incorporated into the polymerase complex. Nevertheless, the polymerase complex assembly kinetics and stability were altered by the incorporation of the C-terminally truncated P7. Using the in vitro assembly system for phi6 nucleocapsids and subsequent infectivity assays, we confirmed that full-length P7 is necessary for the formation of infectious viral particles. Contrary to previous results, we found that P7 must be incorporated into polymerase complexes during shell assembly.
Poranen
Minna M
MM
Institute of Biotechnology, Viikki Biocenter 2, University of Helsinki, Helsinki, Finland. minna.poranen@helsinki.fi
Butcher
Sarah J
SJ
Simonov
Vladimir M
VM
Laurinmäki
Pasi
P
Bamford
Dennis H
DH
D000595:Amino Acid Sequence; D000818:Animals; D017126:Bacteriophage phi 6 / Q000502:physiology*; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D046911:Macromolecular Substances / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D019065:Virus Assembly / Q000502:physiology*
D036022:Capsid Proteins; D046911:Macromolecular Substances; D012330:RNA, Double-Stranded; D012321:DNA-Directed RNA Polymerases