Department of Life Sciences, Ben-Gurion University of the Negev, Be'er Sheva 84105, Israel.
The Department of Chemistry and the National Institute for Biotechnology in the Negev, Ben-Gurion University of the Negev, Be'er Sheva 84105, Israel.
Nucleic Acids Res. 2022 Feb 28;50(4):2143-2156. doi: 10.1093/nar/gkac069.
The coexistence of DNA replication and transcription during S-phase requires their tight coordination to prevent harmful conflicts. While extensive research revealed important mechanisms for minimizing these conflicts and their consequences, little is known regarding how the replication and transcription machinery are coordinated in real-time. Here, we developed a live-cell imaging approach for the real-time monitoring of replisome progression and transcription dynamics during a transcription-replication encounter. We found a wave of partial transcriptional repression ahead of the moving replication fork, which may contribute to efficient fork progression through the transcribed gene. Real-time detection of conflicts revealed their negative impact on both processes, leading to fork stalling or slowdown as well as lower transcription levels during gene replication, with different trade-offs observed in defined subpopulations of cells. Our real-time measurements of transcription-replication encounters demonstrate how these processes can proceed simultaneously while maintaining genomic stability, and how conflicts can arise when coordination is impaired.
在 S 期，DNA 复制和转录的共存需要它们紧密协调，以防止有害冲突。虽然广泛的研究揭示了最小化这些冲突及其后果的重要机制，但对于复制和转录机制如何实时协调知之甚少。在这里，我们开发了一种活细胞成像方法，用于实时监测转录-复制遭遇期间复制体的进展和转录动态。我们发现，在移动的复制叉之前，存在一波部分转录抑制，这可能有助于复制叉有效地通过转录基因前进。实时检测冲突揭示了它们对这两个过程的负面影响，导致复制叉停滞或减速，以及在基因复制过程中转录水平降低，在不同的细胞亚群中观察到不同的权衡。我们对转录-复制遭遇的实时测量表明，这些过程如何在保持基因组稳定性的同时可以同时进行，以及当协调受到破坏时冲突如何产生。
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