tetano
Editor, Senior Moderator
Nat Microbiol. 2016 Mar 21;1:16029. doi: 10.1038/nmicrobiol.2016.29.
[h=1]The role of the priming loop in influenza A virus RNA synthesis.[/h] Te Velthuis AJ[SUP]1,[/SUP][SUP]2[/SUP], Robb NC[SUP]2[/SUP], Kapanidis AN[SUP]2[/SUP], Fodor E[SUP]1[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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 F?rster 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.
PMID: 27572643 DOI: 10.1038/nmicrobiol.2016.29
[PubMed - in process]
[h=1]The role of the priming loop in influenza A virus RNA synthesis.[/h] Te Velthuis AJ[SUP]1,[/SUP][SUP]2[/SUP], Robb NC[SUP]2[/SUP], Kapanidis AN[SUP]2[/SUP], Fodor E[SUP]1[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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 F?rster 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.
PMID: 27572643 DOI: 10.1038/nmicrobiol.2016.29
[PubMed - in process]