@article{179176,
  keywords = {Protein Conformation, RNA, Viral, Transcription, Genetic, DNA Mutational Analysis, Viral Proteins, Virus Replication, Protein Domains, Influenza A virus},
  author = {Aartjan Te Velthuis and Nicole Robb and Achillefs Kapanidis and Ervin Fodor},
  title = {The role of the priming loop in influenza A virus RNA synthesis},
  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 F{\"o}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.  },
  year = {2016},
  journal = {Nat Microbiol},
  volume = {1},
  pages = {16029},
  month = {03/2016},
  issn = {2058-5276},
  doi = {10.1038/nmicrobiol.2016.29},
  language = {eng},
}