Real-time analysis of single influenza virus replication complexes reveals large promoter-dependent differences in initiation dynamics.

Abstract:

:The viral RNA (vRNA) genome of influenza viruses is replicated by the RNA-dependent RNA polymerase (RNAP) via a complementary RNA (cRNA) intermediate. The vRNA promoter can adopt multiple conformations when bound by the RNAP. However, the dynamics, determinants, and biological role of these conformations are unknown; further, little is known about cRNA promoter conformations. To probe the RNA conformations adopted during initial replication, we monitored single, surface-immobilized vRNA and cRNA initiation complexes in real-time. Our results show that, while the 3' terminus of the vRNA promoter exists in dynamic equilibrium between pre-initiation and initiation conformations, the cRNA promoter exhibited very limited dynamics. Two residues in the proximal 3' region of the cRNA promoter (residues absent in the vRNA promoter) allowed the cRNA template strand to reach further into the active site, limiting promoter dynamics. Our results highlight promoter-dependent differences in influenza initiation mechanisms, and advance our understanding of virus replication.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Robb NC,Te Velthuis AJW,Fodor E,Kapanidis AN

doi

10.1093/nar/gkz313

subject

Has Abstract

pub_date

2019-07-09 00:00:00

pages

6466-6477

issue

12

eissn

0305-1048

issn

1362-4962

pii

5481152

journal_volume

47

pub_type

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