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TitleStructural snapshots of actively transcribing influenza polymerase.
Journal, issue, pagesNat. Struct. Mol. Biol., Vol. 26, Issue 6, Page 460-470, Year 2019
Publish dateJun 3, 2019
AuthorsTomas Kouba / Petra Drncová / Stephen Cusack /
PubMed AbstractInfluenza virus RNA-dependent RNA polymerase uses unique mechanisms to transcribe its single-stranded genomic viral RNA (vRNA) into messenger RNA. The polymerase is initially bound to a promoter ...Influenza virus RNA-dependent RNA polymerase uses unique mechanisms to transcribe its single-stranded genomic viral RNA (vRNA) into messenger RNA. The polymerase is initially bound to a promoter comprising the partially base-paired 3' and 5' extremities of the RNA. A short, capped primer, 'cap-snatched' from a nascent host polymerase II transcript, is directed towards the polymerase active site to initiate RNA synthesis. Here we present structural snapshots, as determined by X-ray crystallography and cryo-electron microscopy, of actively initiating influenza polymerase as it transitions towards processive elongation. Unexpected conformational changes unblock the active site cavity to allow establishment of a nine-base-pair template-product RNA duplex before the strands separate into distinct exit channels. Concomitantly, as the template translocates, the promoter base pairs are broken and the template entry region is remodeled. These structures reveal details of the influenza polymerase active site that will help optimize nucleoside analogs or other compounds that directly inhibit viral RNA synthesis.
External linksPubMed:31160782 / Publisher's page
KeywordsVIRAL PROTEIN / Influenza polzmerase / Viral protein / Influenza B Polymerase / RNA dependent RNA polymerase
MethodsEM (single particle) / X-ray diffraction
Resolution2.88 - 3.24 A
Structure data

EMDB-4511:
Influenza B polymerase pre-initiation complex

EMDB-4512:
Influenza B polymerase elongation complex

PDB-6qcs:
Influenza B polymerase pre-initiation complex

PDB-6qct:
Influenza B polymerase elongation complex

PDB-6qcv:
Crystal structure of influenza B polymerase initiation state with capped 14-mer RNA primer and CTP

PDB-6qcw:
Crystal structure of influenza B polymerase initiation state with capped 14-mer RNA primer

PDB-6qcx:
Crystal structure of influenza B polymerase initiation state with capped 15-mer RNA primer

Chemicals

ChemComp-MG:
MAGNESIUM IONMagnesium

ChemComp-PO4:
PHOSPHATE IONPhosphate

ChemComp-CTP:
CYTIDINE-5'-TRIPHOSPHATECytidine triphosphate

ChemComp-POP:
PYROPHOSPHATE 2-Pyrophosphate

Source
  • influenza b virus
  • synthetic construct (others)

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