Structural basis of replication restart primosome proteins involved in DnaB helicase loading

我们对大肠杆菌中参与复制重启的PriB、DnaT和PriC蛋白进行了结构和功能分析。PriB是一种单链DNA （ssDNA）结合蛋白。我们的NMR和FRET结果表明，PriB以两步结合的方式与ssDNA结合，表明PriB与ssDNA之间存在合作结合。我们还对DnaT和pricc进行了域分析。根据DnaT的结构域信息，我们认为DnaT的n端结构域参与了三聚体的形成和与PriB的相互作用，根据核磁共振分析确定的结构，DnaT的c端结构域参与了ssDNA的结合。此外，pricc的结构域信息表明，pricc的c端结构域参与了ssDNA和SSB（单链DNA结合蛋白）的结合。此外，我们利用核磁共振分析确定了PriC的n端结构域。结合这些结果，我们提出了大肠杆菌的复制重启模型。

We performed the structure and function analysis of PriB, DnaT and PriC proteins engaged in replication restart in Escherichia coli. PriB is a single-stranded DNA (ssDNA) binding protein. Our NMR and FRET results showed that PriB bound to ssDNA in two-step binding manner, suggesting the cooperative binding between PriB and ssDNA. We also performed domain analysis of DnaT and PriC. From the domain information of DnaT, we suggested that the N-terminal domain of DnaT was involved in trimer formation and interaction with PriB, and the C-terminal domain of DnaT was involved in ssDNA binding based on the structure determined by NMR analysis. Furthermore, the domain information of PriC suggested that the C-terminal domain of PriC was involved in the ssDNA and SSB (single-stranded DNA binding protein) binding. Additionally, we determined the N-terminal domain of PriC using NMR analysis. Together with these results, we proposed the model of replication restart in Escherichia coli.