Packaging motor from double-stranded RNA bacteriophage phi12 acts as an obligatory passive conduit during transcription.
Double-stranded RNA viruses sequester their genomes within a protein shell, called the polymerase complex. Translocation of ssRNA into (packaging) and out (transcription) of the polymerase complex are essential steps in the life cycle of the dsRNA bacteriophages of the Cystoviridae family (phi6-phi14). Both processes require a viral molecular motor P4, an NTPase, which bears structural and functional similarities to hexameric helicases. In effect, switching between the packaging and the transcription mode requires the translocation direction of the P4 motor to reverse. However, the mechanism of the reversal remains elusive. Here we characterize the P4 protein from bacteriophage phi12 and exploit its purine nucleotide specificity to delineate P4 role in transcription. The results indicate that while P4 actively translocates RNA during packaging it acts as a passive conduit for RNA export. The directionality switching is accomplished via the regulation of P4 NTPase activity within the polymerase core.
Kainov
Denis E
DE
Institute of Biotechnology and Faculty of Biological Science, Viikki Biocenter, University of Helsinki, PO Box 65, Viikinkaari 1, Helsinki FIN-00014, Finland.
Lísal
Jirí
J
Bamford
Dennis H
DH
Tuma
Roman
R
D017126:Bacteriophage phi 6 / Q000235:genetics; D001692:Biological Transport; D017910:Cystoviridae / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D020409:Molecular Motor Proteins / Q000378:metabolism; D043583:Nucleoside-Triphosphatase / Q000378:metabolism*; D012313:RNA / Q000378:metabolism; D020365:RNA Helicases / Q000378:metabolism; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000378:metabolism*; D019065:Virus Assembly
D020409:Molecular Motor Proteins; D012330:RNA, Double-Stranded; D014764:Viral Proteins; D012313:RNA; D012321:DNA-Directed RNA Polymerases; D043583:Nucleoside-Triphosphatase; D020365:RNA Helicases