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

Chun-Ying Lee, Christina McNerney, Kevin Ma, Walter Zhao, Ashley Wang and Sua Myong ()
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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

Nature Communications, 2020, vol. 11, issue 1, 1-15

Abstract: 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.

Date: 2020
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DOI: 10.1038/s41467-020-17176-7

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