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- W2913228809 abstract "G-quadruplex (G4) is a noncanonical secondary structure that forms in G-rich sequences of DNA or RNA. G4 has been implicated in gene regulation and human disease. Current evidences indicate that formation of G4 in DNA and RNA modulates replication, transcription and translation. It's hypothesized that G4 can act as a barrier to repress DNA and RNA polymerase (RNAP), but it can also facilitate the same process by keeping DNA unwound, for example. However, how G4 forming sequence impacts enzymatic process of polymerase is still unclear. Here, we sought to investigate if and how G4 forming sequences at 5’-UTR alters transcription by comparing G4 sequences inserted in template vs. non-template orientation. Using in vitro transcription assay coupled with molecular beacon (dequenching probe), we found that all tested G4 sequences positioned in template strand reduced, but the same G4 in non-template strand enhanced mRNA production. To dissect various stages involved in transcription, we devised four sequential single molecule assays which enable measurement of T7 RNAP binding, initiation, elongation, mRNA product formation. Our result shows that while binding and initiation are similar, the proportion of successful elongation and the final mRNA yield is lower when G4 is on template and higher when G4 is on non-template. In addition, the duration of transcription bubble at the G4 sequence is shorter on template than in non-template, suggesting a potential secondary structure such as G-loop or R-loop modulating the transcriptional output at the G4 site. Our study provides a detailed kinetic map of transcription of G4 bearing template in which G4 orientation plays a major role in determining the transcriptional output." @default.
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- W2913228809 date "2019-02-01" @default.
- W2913228809 modified "2023-10-08" @default.
- W2913228809 title "G-Quadruplex Forming Sequence Modulated Transcription" @default.
- W2913228809 doi "https://doi.org/10.1016/j.bpj.2018.11.1154" @default.
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