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TitleStructural Insights into Bunyavirus Replication and Its Regulation by the vRNA Promoter.
Journal, issue, pagesCell, Vol. 161, Issue 6, Page 1267-1279, Year 2015
Publish dateJun 4, 2015
AuthorsPiotr Gerlach / Hélène Malet / Stephen Cusack / Juan Reguera /
PubMed AbstractSegmented negative-strand RNA virus (sNSV) polymerases transcribe and replicate the viral RNA (vRNA) within a ribonucleoprotein particle (RNP). We present cryo-EM and X-ray structures of, ...Segmented negative-strand RNA virus (sNSV) polymerases transcribe and replicate the viral RNA (vRNA) within a ribonucleoprotein particle (RNP). We present cryo-EM and X-ray structures of, respectively, apo- and vRNA bound La Crosse orthobunyavirus (LACV) polymerase that give atomic-resolution insight into how such RNPs perform RNA synthesis. The complementary 3' and 5' vRNA extremities are sequence specifically bound in separate sites on the polymerase. The 5' end binds as a stem-loop, allosterically structuring functionally important polymerase active site loops. Identification of distinct template and product exit tunnels allows proposal of a detailed model for template-directed replication with minimal disruption to the circularised RNP. The similar overall architecture and vRNA binding of monomeric LACV to heterotrimeric influenza polymerase, despite high sequence divergence, suggests that all sNSV polymerases have a common evolutionary origin and mechanism of RNA synthesis. These results will aid development of replication inhibitors of diverse, serious human pathogenic viruses.
External linksCell / PubMed:26004069 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.57 - 8.3 Å
Structure data

EMDB-2930:
Cryo-electron microscopy map of La Crosse orthobunyavirus polymerase (apo form, residues 1-1750)
Method: EM (single particle) / Resolution: 8.3 Å

PDB-5amq:
Structure of the La Crosse Bunyavirus polymerase in complex with the 3' and 5' viral RNA
Method: X-RAY DIFFRACTION / Resolution: 3.0 Å

PDB-5amr:
Structure of the La Crosse Bunyavirus polymerase in complex with the 3' viral RNA
Method: X-RAY DIFFRACTION / Resolution: 2.57 Å

Chemicals

ChemComp-HOH:
WATER / Water

Source
  • La Crosse virus
  • bunyavirus la crosse
KeywordsHYDROLASE / POLYMERASE / RNADRNAPOL / BUNYAVIRUS / RNA

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