2RU8

DnaT C-terminal domain

2RU8 の概要

• エントリーDOI: 10.2210/pdb2ru8/pdb
• NMR情報: BMRB: 11549
• 分子名称: Primosomal protein 1 (1 entity in total)
• 機能のキーワード: primosome, replication restart, dnat, dna binding, replication
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 11237.62

構造登録者

Abe, Y.,Tani, J.,Fujiyama, S.,Urabe, M.,Sato, K.,Aramaki, T.,Katayama, T.,Ueda, T. (登録日: 2014-01-29, 公開日: 2014-10-08, 最終更新日: 2024-05-15)

主引用文献

Fujiyama, S.,Abe, Y.,Tani, J.,Urabe, M.,Sato, K.,Aramaki, T.,Katayama, T.,Ueda, T.
Structure and mechanism of the primosome protein DnaT-functional structures for homotrimerization, dissociation of ssDNA from the PriB·ssDNA complex, and formation of the DnaT·ssDNA complex.
Febs J., 281:5356-5370, 2014

PubMed Abstract: In Escherichia coli, the primosome plays an essential role in replication restart after dissociation of replisomes at the damaged replication fork. As well as PriA and PriB, DnaT, an ssDNA-binding protein, is a key member of the primosome. In this study, limited proteolysis indicated that E. coli DnaT was composed of two domains, consistent with the results of recent studies using Klebsiella pneumonia DnaT. We also found that a specific 24-residue region (Phe42-Asp66) in the N-terminal domain (1-88) was crucial for DnaT trimerization. Moreover, we determined the structure of the DnaT C-terminal domain (89-179) by NMR spectroscopy. This domain included three α-helices and a long flexible C-terminal tail, similar to the C-terminal subdomain of the AAA+ ATPase family. The neighboring histidines, His136 and His137, at a position corresponding to the AAA+ sensor II motif, were suggested to form an ssDNA-binding site. Furthermore, we found that the acidic linker between the two domains had an activity for dissociating ssDNA from the PriB·ssDNA complexes in a manner supported by the conserved acidic residues Asp70 and Glu76. Thus, these findings provide a novel structural basis for understanding the mechanism of DnaT in exposure of ssDNA and reloading of the replicative helicase at the stalled replication fork.

PubMed: 25265331
DOI: 10.1111/febs.13080

実験手法

SOLUTION NMR