Program in Cellular and Molecular Medicine, Boston Children's Hospital and Harvard Medical School, Boston, MA, United States.
Program in Cellular Molecular Developmental Biology and Biophysics, Johns Hopkins University, Baltimore, MD, United States.
Methods Enzymol. 2024;695:71-88. doi: 10.1016/bs.mie.2024.01.001. Epub 2024 Jan 16.
Potential G-quadruplex forming sequences (PQS) are enriched in cancer-related genes and immunoglobulin class-switch recombination. They are prevalent in the 5'UTR of transcriptionally active genes, thereby contributing to the regulation of gene expression. We and others previously demonstrated that the PQS located in the non-template strand leads to an R-loop formation followed by a G-quadruplex (G4) formation during transcription. These structural changes increase mRNA production. Here, we present how single-molecule technique was used to observe cotranscriptional G4 and R-loop formation and to examine the impact on transcription, particularly for the initiation and elongation stages.
潜在的 G-四链体形成序列(PQS)在与癌症相关的基因和免疫球蛋白类别转换重组中富集。它们在转录活跃基因的 5'UTR 中普遍存在,从而有助于基因表达的调控。我们和其他人之前已经证明,位于非模板链上的 PQS 会导致转录过程中形成 R 环,随后形成 G-四链体(G4)。这些结构变化会增加 mRNA 的产生。在这里,我们介绍了如何使用单分子技术观察共转录 G4 和 R 环的形成,并研究其对转录的影响,特别是对起始和延伸阶段的影响。