Initiation, elongation, and realignment during influenza virus mRNA synthesis

Aartjan J.W. te Velthuis, Judith Oymans

Research output: Contribution to journalArticleAcademicpeer-review

11 Citations (Scopus)

Abstract

The RNA-dependent RNA polymerase (RdRp) of the influenza A virus replicates and transcribes the viral genome segments in the nucleus of the host cell. To transcribe these viral genome segments, the RdRp "snatches" capped RNA oligonucleotides from nascent host cell mRNAs and aligns these primers to the ultimate or penultimate nucleotide of the segments for the initiation of viral mRNA synthesis. It has been proposed that this initiation process is not processive and that the RdRp uses a prime-realign mechanism during transcription. Here we provide in vitro evidence for the existence of this transcriptional prime-realign mechanism but show that it functions efficiently only for primers that are short or cannot stably base pair with the template. In addition, we demonstrate that transcriptional elongation is dependent on the priming loop of the PB1 subunit of the RdRp. We propose that the prime-realign mechanism may be used to rescue abortive transcription initiation events or cope with sequence variation among primers. Overall, these observations advance our mechanistic understanding of how influenza A virus initiates transcription correctly and efficiently.
Original languageEnglish
Article numbere01775-17
JournalJournal of Virology
Volume92
Issue number3
DOIs
Publication statusPublished - 1 Feb 2018

Fingerprint

RNA-directed RNA polymerase
RNA Replicase
Orthomyxoviridae
Messenger RNA
synthesis
transcription (genetics)
Viral Genome
Influenza A virus
genome
oligonucleotides
Cell Nucleus
Oligonucleotides
Base Pairing
Nucleotides
nucleotides
RNA
cells

Keywords

  • Influenza A virus
  • Priming loop
  • Realignment
  • Replication
  • RNA-dependent RNA polymerase
  • Transcription
  • Viral transcription

Cite this

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title = "Initiation, elongation, and realignment during influenza virus mRNA synthesis",
abstract = "The RNA-dependent RNA polymerase (RdRp) of the influenza A virus replicates and transcribes the viral genome segments in the nucleus of the host cell. To transcribe these viral genome segments, the RdRp {"}snatches{"} capped RNA oligonucleotides from nascent host cell mRNAs and aligns these primers to the ultimate or penultimate nucleotide of the segments for the initiation of viral mRNA synthesis. It has been proposed that this initiation process is not processive and that the RdRp uses a prime-realign mechanism during transcription. Here we provide in vitro evidence for the existence of this transcriptional prime-realign mechanism but show that it functions efficiently only for primers that are short or cannot stably base pair with the template. In addition, we demonstrate that transcriptional elongation is dependent on the priming loop of the PB1 subunit of the RdRp. We propose that the prime-realign mechanism may be used to rescue abortive transcription initiation events or cope with sequence variation among primers. Overall, these observations advance our mechanistic understanding of how influenza A virus initiates transcription correctly and efficiently.",
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Initiation, elongation, and realignment during influenza virus mRNA synthesis. / te Velthuis, Aartjan J.W.; Oymans, Judith.

In: Journal of Virology, Vol. 92, No. 3, e01775-17, 01.02.2018.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Initiation, elongation, and realignment during influenza virus mRNA synthesis

AU - te Velthuis, Aartjan J.W.

AU - Oymans, Judith

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N2 - The RNA-dependent RNA polymerase (RdRp) of the influenza A virus replicates and transcribes the viral genome segments in the nucleus of the host cell. To transcribe these viral genome segments, the RdRp "snatches" capped RNA oligonucleotides from nascent host cell mRNAs and aligns these primers to the ultimate or penultimate nucleotide of the segments for the initiation of viral mRNA synthesis. It has been proposed that this initiation process is not processive and that the RdRp uses a prime-realign mechanism during transcription. Here we provide in vitro evidence for the existence of this transcriptional prime-realign mechanism but show that it functions efficiently only for primers that are short or cannot stably base pair with the template. In addition, we demonstrate that transcriptional elongation is dependent on the priming loop of the PB1 subunit of the RdRp. We propose that the prime-realign mechanism may be used to rescue abortive transcription initiation events or cope with sequence variation among primers. Overall, these observations advance our mechanistic understanding of how influenza A virus initiates transcription correctly and efficiently.

AB - The RNA-dependent RNA polymerase (RdRp) of the influenza A virus replicates and transcribes the viral genome segments in the nucleus of the host cell. To transcribe these viral genome segments, the RdRp "snatches" capped RNA oligonucleotides from nascent host cell mRNAs and aligns these primers to the ultimate or penultimate nucleotide of the segments for the initiation of viral mRNA synthesis. It has been proposed that this initiation process is not processive and that the RdRp uses a prime-realign mechanism during transcription. Here we provide in vitro evidence for the existence of this transcriptional prime-realign mechanism but show that it functions efficiently only for primers that are short or cannot stably base pair with the template. In addition, we demonstrate that transcriptional elongation is dependent on the priming loop of the PB1 subunit of the RdRp. We propose that the prime-realign mechanism may be used to rescue abortive transcription initiation events or cope with sequence variation among primers. Overall, these observations advance our mechanistic understanding of how influenza A virus initiates transcription correctly and efficiently.

KW - Influenza A virus

KW - Priming loop

KW - Realignment

KW - Replication

KW - RNA-dependent RNA polymerase

KW - Transcription

KW - Viral transcription

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DO - 10.1128/JVI.01775-17

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JO - Journal of Virology

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