デルフト工科大学の研究者が、DNA複製のモーターに新たな光を当てる(TU Delft researchers shed new light on the motor of DNA replication)

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2023-04-14 オランダ・デルフト工科大学(TUDelft)

オランダ・デルフト工科大学の研究チームは、DNA複製のための分子モーターであるCMGの運動を前例のない解像度で組み立て、観察するための新しい手法を開発した。この手法により、DNA複製研究の新しい発見が可能になる。
CMGは、DNAの二重螺旋を分離して複製する重要な役割を持っている。また、ATPが欠落すると、CMGはDNA上をランダムに移動することがわかった。
これらの発見は、遺伝子疾患やがんの原因となるDNA複製の誤りについての理解を深めることにつながる。

<関連情報>

真核生物の複製ヘリカーゼの活性化時に、ヌクレオチド結合が拡散を停止させる Nucleotide binding halts diffusion of the eukaryotic replicative helicase during activation

Daniel Ramírez Montero,Humberto Sánchez,Edo van Veen,Theo van Laar,Belén Solano,John F. X. Diffley & Nynke H. Dekker
Nature Communications  Published:14 April 2023
DOI:https://doi.org/10.1038/s41467-023-37093-9

figure 1

Abstract

The eukaryotic replicative helicase CMG centrally orchestrates the replisome and leads the way at the front of replication forks. Understanding the motion of CMG on the DNA is therefore key to our understanding of DNA replication. In vivo, CMG is assembled and activated through a cell-cycle-regulated mechanism involving 36 polypeptides that has been reconstituted from purified proteins in ensemble biochemical studies. Conversely, single-molecule studies of CMG motion have thus far relied on pre-formed CMG assembled through an unknown mechanism upon overexpression of individual constituents. Here, we report the activation of CMG fully reconstituted from purified yeast proteins and the quantification of its motion at the single-molecule level. We observe that CMG can move on DNA in two ways: by unidirectional translocation and by diffusion. We demonstrate that CMG preferentially exhibits unidirectional translocation in the presence of ATP, whereas it preferentially exhibits diffusive motion in the absence of ATP. We also demonstrate that nucleotide binding halts diffusive CMG independently of DNA melting. Taken together, our findings support a mechanism by which nucleotide binding allows newly assembled CMG to engage with the DNA within its central channel, halting its diffusion and facilitating the initial DNA melting required to initiate DNA replication.

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