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Single-molecule FRET Studies on DNA Mismatch Repair

机译：DNA错配修复的单分子FRET研究

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DNA mismatch repair (MMR) is the process that corrects misincorporation errors introduced by DNA polymerases during replication. MMR is also associated with other biological processes such as the suppression of recombination between partially homologous sequences (homeologous recombination) and DNA-damage signaling. Mutations of the human MMR genes are the cause of Lynch syndrome, also known as hereditary nonpolyposis colorectal cancer (LS/HNPCC). The detailed mechanism of MMR components in these biological processes remains enigmatic. MMR that is coupled to replication is an excision-resynthesis reaction. It is initiated at a distant strand scission that is 100’s to 1000’s of bp from the mismatch and the excision tract extends from that strand scission to just past the mismatch. MutS and MutL are unique core components of MMR that recognize a mismatch and initiate the excision reaction. Additional components include the replication clamp (β-clamp in prokaryotes and PCNA in eukaryotes), the replication clamp loader (γδδ’ in prokaryotes and RFC in eukaryotes), an exonuclease (Exo1, ExoX, RecJ, ExoVII in prokaryotes and EXOI in eukaryotes), and single strand binding protein (SSB in prokaryotes and RPA in eukaryotes). Recently single molecule FRET/Fluorescent Tracking (smFRET/FT) studies have made a significant impact on understanding the MMR process. More extensive future smFRET/FT studies are expected to further detail the MMR mechanism. This review is intended to offer a guide to applying smFRET/FT studies to understand the entire MMR process, by pinpointing key questions and poorly understood phenomena. We summarize recent smFRET/FT results and address major issues in the application of the smFRET/FT system.
机译：DNA错配修复（MMR）是纠正DNA聚合酶在复制过程中引入的错误掺入错误的过程。 MMR还与其他生物学过程相关，例如抑制部分同源序列之间的重组（同源重组）和DNA损伤信号传导。人类MMR基因的突变是Lynch综合征的原因，也称为遗传性非息肉性结直肠癌（LS / HNPCC）。这些生物过程中MMR组件的详细机制仍然是个谜。与复制偶联的MMR是切除再合成反应。它是在距离错配100到1000 bp的远距离链断裂处开始的，并且切割线从该链断裂延伸到刚刚超过错配。 MutS和MutL是MMR的独特核心组件，可识别错配并启动切除反应。其他组件包括复制钳（原核生物中的β钳和真核生物中的PCNA），复制钳装载器（原核生物中的γδδ'和真核生物中的RFC），外切核酸酶（原核生物中的Exo1，ExoX，RecJ，ExoVII和真核生物中的EXOI） ，和单链结合蛋白（原核生物中的SSB和真核生物中的RPA）。最近，单分子FRET /荧光跟踪（smFRET / FT）研究对理解MMR过程产生了重大影响。预期将来会进行更广泛的smFRET / FT研究，以进一步详述MMR机制。这篇综述旨在通过指出关键问题和理解不足的现象，为应用smFRET / FT研究提供指南，以了解整个MMR过程。我们总结了最近的smFRET / FT结果，并解决了smFRET / FT系统应用中的主要问题。

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