Journal
NUCLEIC ACIDS RESEARCH
Volume 47, Issue 12, Pages 6466-6477Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkz313
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Funding
- Royal Society Dorothy Hodgkin Research Fellowship [DKR00620]
- Wellcome Trust [206579/Z/17/Z, 110164/Z/15/Z]
- Royal Society [206579/Z/17/Z]
- Marmaduke Shield Fund award [17.37(r)]
- Isaac Newton Trust [17.37(r)]
- Medical Research Council [MR/K000241/1, MR/R009945/1, MR/N010744/1]
- Oxford's RCUK Open Access Block Grant
- Wellcome Trust [206579/Z/17/Z] Funding Source: Wellcome Trust
- MRC [MR/R009945/1, MR/K000241/1, MR/N010744/1] Funding Source: UKRI
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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.
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