Direct observation of backtracking by influenza A and B polymerases upon consecutive incorporation of the nucleoside analog T1106:Cell Reports

Link:
Autor/in:
Erscheinungsjahr:
2023
Medientyp:
Text
Schlagworte:
  • antiviral drug
  • backtracking
  • cap-dependent transcription
  • CP: Molecular biology
  • influenza virus
  • molecular dynamics
  • nucleoside analog
  • RNA-dependent RNA polymerase
  • single-particle cryogenic electron microscopy
  • T1106
  • T705
  • Antiviral Agents
  • DNA-Directed RNA Polymerases
  • Humans
  • Influenza A Virus, H7N9 Subtype
  • Influenza, Human
  • Nucleosides
  • Nucleotides
  • antivirus agent
  • DNA polymerase
  • nucleotide
  • pyrazinamide
  • RNA directed DNA polymerase
  • RNA directed RNA polymerase
  • RNA polymerase
  • unclassified drug
  • DNA directed RNA polymerase
  • nucleoside
  • affinity chromatography
  • Article
  • binding site
  • classification
  • conformational transition
  • controlled study
  • cryoelectron microscopy
  • crystal structure
  • density functional theory
  • DNA binding
  • electron microscopy
  • genetic transcription
  • human
  • human cell
  • hydrogen bond
  • influenza A
  • influenza A (H7N9)
  • influenza B
  • Influenza virus
  • molecular biology
  • molecular cloning
  • molecular docking
  • nucleosome
  • nucleotide sequence
  • protein purification
  • protein secondary structure
  • refraction index
  • RNA binding
  • RNA sequence
  • RNA synthesis
  • sequence alignment
  • site directed mutagenesis
  • static electricity
  • three-dimensional imaging
  • transcription termination
  • Trichoplusia ni
  • ultrasound
  • validation process
  • virus replication
  • wobble base pair
  • X ray crystallography
  • influenza
  • Influenza A virus (H7N9)
  • metabolism
Beschreibung:
  • The antiviral pseudo-base T705 and its de-fluoro analog T1106 mimic adenine or guanine and can be competitively incorporated into nascent RNA by viral RNA-dependent RNA polymerases. Although dispersed, single pseudo-base incorporation is mutagenic, consecutive incorporation causes polymerase stalling and chain termination. Using a template encoding single and then consecutive T1106 incorporation four nucleotides later, we obtained a cryogenic electron microscopy structure of stalled influenza A/H7N9 polymerase. This shows that the entire product-template duplex backtracks by 5 nt, bringing the singly incorporated T1106 to the +1 position, where it forms an unexpected T1106:U wobble base pair. Similar structures show that influenza B polymerase also backtracks after consecutive T1106 incorporation, regardless of whether prior single incorporation has occurred. These results give insight into the unusual mechanism of chain termination by pyrazinecarboxamide base analogs. Consecutive incorporation destabilizes the proximal end of the product-template duplex, promoting irreversible backtracking to a more energetically favorable overall configuration. © 2022 The Author(s)
Lizenz:
  • info:eu-repo/semantics/closedAccess
Quellsystem:
Forschungsinformationssystem der UHH
Interne Metadaten
Quelldatensatz
oai:www.edit.fis.uni-hamburg.de:publications/a265e18b-67a7-4eb4-9b0f-fb3aa544bbc1