Pre-initiation and elongation structures of full-length La Crosse virus polymerase reveal functionally important conformational changes.

Nature Communications
Benoît ArragainHélène Malet

Abstract

Bunyavirales is an order of segmented negative-strand RNA viruses comprising several life-threatening pathogens against which no effective treatment is currently available. Replication and transcription of the RNA genome constitute essential processes performed by the virally encoded multi-domain RNA-dependent RNA polymerase. Here, we describe the complete high-resolution cryo-EM structure of La Crosse virus polymerase. It reveals the presence of key protruding C-terminal domains, notably the cap-binding domain, which undergoes large movements related to its role in transcription initiation, and a zinc-binding domain that displays a fold not previously observed. We capture the polymerase structure at pre-initiation and elongation states, uncovering the coordinated movement of the priming loop, mid-thumb ring linker and lid domain required for the establishment of a ten-base-pair template-product RNA duplex before strand separation into respective exit tunnels. These structural details and the observed dynamics of key functional elements will be instrumental for structure-based development of polymerase inhibitors.

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Citations

Jan 27, 2021·Nature Reviews. Microbiology·Aartjan J W Te VelthuisErvin Fodor
Jun 3, 2021·Viruses·Richard KormelinkCecile Desbiez
Aug 29, 2021·Viruses·Kristina MeierMaria Rosenthal
Aug 26, 2021·Journal of Virology·Jesse D Pyle, Sean P J Whelan

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Datasets Mentioned

BETA
EF485038.1

Methods Mentioned

BETA
X-ray
gel
size exclusion chromatography

Software Mentioned

cryoSPARC
MUSCLE
PHASER
DALI
Cryo
Phenix
EM
Relion
STARANISO
Chimera

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