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

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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.