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R-loop induced G-quadruplex in non-template promotes transcription by successive R-loop formation

Author

Listed:
  • Chun-Ying Lee

    (Johns Hopkins University)

  • Christina McNerney

    (Johns Hopkins University)

  • Kevin Ma

    (Johns Hopkins University)

  • Walter Zhao

    (Johns Hopkins University)

  • Ashley Wang

    (Johns Hopkins University)

  • Sua Myong

    (Johns Hopkins University
    University of Illinois)

Abstract

G-quadruplex (G4) is a noncanonical secondary structure of DNA or RNA which can enhance or repress gene expression, yet the underlying molecular mechanism remains uncertain. Here we show that when positioned downstream of transcription start site, the orientation of potential G4 forming sequence (PQS), but not the sequence alters transcriptional output. Ensemble in vitro transcription assays indicate that PQS in the non-template increases mRNA production rate and yield. Using sequential single molecule detection stages, we demonstrate that while binding and initiation of T7 RNA polymerase is unchanged, the efficiency of elongation and the final mRNA output is higher when PQS is in the non-template. Strikingly, the enhanced elongation arises from the transcription-induced R-loop formation, which in turn generates G4 structure in the non-template. The G4 stabilized R-loop leads to increased transcription by a mechanism involving successive rounds of R-loop formation.

Suggested Citation

  • Chun-Ying Lee & Christina McNerney & Kevin Ma & Walter Zhao & Ashley Wang & Sua Myong, 2020. "R-loop induced G-quadruplex in non-template promotes transcription by successive R-loop formation," Nature Communications, Nature, vol. 11(1), pages 1-15, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-17176-7
    DOI: 10.1038/s41467-020-17176-7
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    Cited by:

    1. Chun-Ying Lee & Meera Joshi & Ashley Wang & Sua Myong, 2024. "5′UTR G-quadruplex structure enhances translation in size dependent manner," Nature Communications, Nature, vol. 15(1), pages 1-13, December.
    2. Heonjoon Lee & Tian Xie & Byunghwa Kang & Xinjie Yu & Samuel W. Schaffter & Rebecca Schulman, 2024. "Plug-and-play protein biosensors using aptamer-regulated in vitro transcription," Nature Communications, Nature, vol. 15(1), pages 1-11, December.

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