Protein P7 of phage phi6 RNA polymerase complex, acquiring of RNA packaging activity by in vitro assembly of the purified protein onto deficient particles.
The RNA polymerase complex of double-stranded RNA bacteriophage phi6 is composed of four proteins, P1, P2, P4 and P7. These four proteins are capable of performing all the functions required for the replication of the double-stranded RNAs of the phi6 genome. The polymerase complex containing the three genomic dsRNA segments is the core particle of the phi6 virion. In this study purified protein P7 was found to form highly asymmetric dimers. Using polyclonal anti-P7 antibody, P7 was shown to be accessible on the surface of the nucleocapsid. Treatment of nucleocapsids with polyclonal anti-P7 antibody released coat protein P8 with ensuing activation of the plus strand RNA synthesis from the resulting core particles. Purified P7 could be assembled onto particles lacking P7 and particles lacking both P2 (RNA polymerase) and P7. In both cases RNA packaging activity was acquired. Assembly of P7 onto deficient particles took place also in the absence of host proteins. Protein P7 is known to be necessary for stable packaging of the three genomic phi6 plus strand RNAs into preformed polymerase complex particles. Additionally, protein P7 seems to be involved in the regulation of plus strand synthesis (i.e. transcription) as a fidelity factor. Particles lacking protein P7 produce anomalous size transcripts. Analysis of the polymerase complex stability revealed that proteins P2, P4 and P7 are independently associated with the major structural protein P1. The number of P7 molecules in one virion was estimated to be 60 and a location at the 5-fold symmetry position is proposed.
Juuti
J T
JT
Department of Biosciences, University of Helsinki, Finland.
Bamford
D H
DH
D017126:Bacteriophage phi 6 / Q000201:enzymology / Q000254:growth & development*; D002213:Capsid / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019251:Nucleocapsid / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D012367:RNA, Viral / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000737:chemistry
D012367:RNA, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; D012321:DNA-Directed RNA Polymerases