The role of the priming loop in influenza A virus RNA synthesis
TLDR
Analysis of individual residues in the tip of the β-hairpin shows that PB1 proline 651 is critical for efficient RNA synthesis in vitro and in cell culture, which advances the understanding of influenza A virus RNA synthesis and identifies the initiation platform of viral replication.Abstract:
RNA-dependent RNA polymerases (RdRps) are used by RNA viruses to replicate and transcribe their RNA genomes(1). They adopt a closed, right-handed fold with conserved subdomains called palm, fingers and thumb(1,2). Conserved RdRp motifs A-F coordinate the viral RNA template, NTPs and magnesium ions to facilitate nucleotide condensation(1). For the initiation of RNA synthesis, most RdRps use either a primer-dependent or de novo mechanism(3). The influenza A virus RdRp, in contrast, uses a capped RNA oligonucleotide to initiate transcription, and a combination of terminal and internal de novo initiation for replication(4). To understand how the influenza A virus RdRp coordinates these processes, we analysed the function of a thumb subdomain β-hairpin using initiation, elongation and single-molecule Forster resonance energy transfer (sm-FRET) assays. Our data indicate that this β-hairpin is essential for terminal initiation during replication, but not necessary for internal initiation and transcription. Analysis of individual residues in the tip of the β-hairpin shows that PB1 proline 651 is critical for efficient RNA synthesis in vitro and in cell culture. Overall, this work advances our understanding of influenza A virus RNA synthesis and identifies the initiation platform of viral replication.read more
Citations
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Influenza virus RNA polymerase: insights into the mechanisms of viral RNA synthesis.
TL;DR: Current knowledge of the structure of the influenza virus RNA polymerase is discussed, and insights that have been gained into the molecular mechanisms of viral transcription and replication, and their regulation by viral and host factors are discussed.
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RNA Dependent RNA Polymerases: Insights from Structure, Function and Evolution
TL;DR: An overview of the structural aspects of RdRps and their complexes from the Group III, IV and V viruses and their structure-based phylogeny is provided.
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Structural insights into RNA synthesis by the influenza virus transcription-replication machine.
TL;DR: This review focusses on the new insights that recent crystal structures have given into the detailed molecular mechanisms by which the polymerase performs both transcription and replication of the vRNA genome.
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Structures of influenza A virus RNA polymerase offer insight into viral genome replication
Haitian Fan,Alexander P Walker,Loic Carrique,J.R. Keown,I. Serna Martin,I. Serna Martin,D. Karia,Jane Sharps,Narin Hengrung,Narin Hengrung,Els Pardon,Jan Steyaert,Jonathan M. Grimes,Ervin Fodor +13 more
TL;DR: This study provides high-resolution structures of medically relevant FluPolA, as well as insights into the replication mechanisms of the viral RNA genome, and identifies sites in Flu PolA that could be targeted in the development of antiviral drugs.
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Nucleosides for the treatment of respiratory RNA virus infections
TL;DR: The current medical need resulting from respiratory infections caused by RNA viruses is described, which justifies drug discovery efforts to identify new therapeutic agents and summarizes past and current efforts to develop nucleoside and nucleotide analogs as antiviral agents against respiratory virus infections.
References
More filters
Journal ArticleDOI
Clustal W and Clustal X version 2.0
Mark A. Larkin,Gordon Blackshields,Nigel P. Brown,R. Chenna,Paul A. McGettigan,Hamish McWilliam,Franck Valentin,Iain M. Wallace,Andreas Wilm,Rodrigo Lopez,J.D. Thompson,Toby J. Gibson,Desmond G. Higgins +12 more
TL;DR: The Clustal W and ClUSTal X multiple sequence alignment programs have been completely rewritten in C++ to facilitate the further development of the alignment algorithms in the future and has allowed proper porting of the programs to the latest versions of Linux, Macintosh and Windows operating systems.
Journal ArticleDOI
Deciphering key features in protein structures with the new ENDscript server
Xavier Robert,Patrice Gouet +1 more
TL;DR: This major upgrade has been fully re-engineered to enhance speed, accuracy and usability with interactive 3D visualization of ENDscript 2 and ESPript 3 to handle a large number of data with reduced computation time.
Journal ArticleDOI
The Activated Complex in Chemical Reactions
TL;DR: In this paper, the probability of the activated state is calculated using ordinary statistical mechanics, and the probability multiplied by the rate of decomposition gives the specific rate of reaction, and necessary conditions for general statistical treatment to reduce to the usual kinetic treatment are given.
Journal ArticleDOI
Amino acid–base interactions: a three-dimensional analysis of protein–DNA interactions at an atomic level
TL;DR: The majority of amino acid-base interactions observed follow general principles that apply across all protein-DNA complexes, although there are individual exceptions.
Journal ArticleDOI
Crystal structure of an ihf-dna complex : a protein-induced dna u-turn
TL;DR: The crystal structure of IHF complexed with 35 bp of DNA is reported, providing evidence for the importance of a narrow minor groove in IHF's binding sequence.