A Structure-Based Model for the Complete Transcription Cycle of Influenza Polymerase.
Catalytic Domain
Computer Simulation
Cryoelectron Microscopy
/ methods
Genome, Viral
/ genetics
Humans
Influenza A virus
/ genetics
Influenza, Human
/ genetics
Nucleotidyltransferases
/ metabolism
RNA, Messenger
/ metabolism
RNA, Viral
/ metabolism
RNA-Dependent RNA Polymerase
/ genetics
Structure-Activity Relationship
Transcription, Genetic
/ genetics
Virus Replication
/ genetics
Influenza virus
RNA-dependent RNA polymerase
cap-snatching
cryo-electron microscopy
end binding site
poly-adenylation
secondary 3’
transcription
Journal
Cell
ISSN: 1097-4172
Titre abrégé: Cell
Pays: United States
ID NLM: 0413066
Informations de publication
Date de publication:
14 05 2020
14 05 2020
Historique:
received:
04
11
2019
revised:
18
02
2020
accepted:
26
03
2020
pubmed:
19
4
2020
medline:
5
1
2021
entrez:
19
4
2020
Statut:
ppublish
Résumé
Influenza polymerase uses unique mechanisms to synthesize capped and polyadenylated mRNAs from the genomic viral RNA (vRNA) template, which is packaged inside ribonucleoprotein particles (vRNPs). Here, we visualize by cryoelectron microscopy the conformational dynamics of the polymerase during the complete transcription cycle from pre-initiation to termination, focusing on the template trajectory. After exiting the active site cavity, the template 3' extremity rebinds into a specific site on the polymerase surface. Here, it remains sequestered during all subsequent transcription steps, forcing the template to loop out as it further translocates. At termination, the strained connection between the bound template 5' end and the active site results in polyadenylation by stuttering at uridine 17. Upon product dissociation, further conformational changes release the trapped template, allowing recycling back into the pre-initiation state. Influenza polymerase thus performs transcription while tightly binding to and protecting both template ends, allowing efficient production of multiple mRNAs from a single vRNP.
Identifiants
pubmed: 32304664
pii: S0092-8674(20)30389-5
doi: 10.1016/j.cell.2020.03.061
pii:
doi:
Substances chimiques
RNA, Messenger
0
RNA, Viral
0
Nucleotidyltransferases
EC 2.7.7.-
RNA-Dependent RNA Polymerase
EC 2.7.7.48
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
877-893.e21Informations de copyright
Copyright © 2020 Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Interests S.C., T.K., and P.D. have a patent application related to this work.