7U82

Human DNA polymerase eta-DNA-dGMPNPP ternary mismatch complex in 1.0 mM Mn2+ for 600s

7U82 の概要

• エントリーDOI: 10.2210/pdb7u82/pdb
• 分子名称: DNA polymerase eta, DNA (5'-D(*CP*AP*TP*TP*AP*TP*GP*AP*CP*GP*CP*T)-3'), DNA (5'-D(*AP*GP*CP*GP*TP*CP*AP*T)-3'), ... (7 entities in total)
• 機能のキーワード: polymerase, transferase-dna complex, transferase/dna
• 由来する生物種: Homo sapiens (human); 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 55481.98

構造登録者

Chang, C.,Gao, Y. (登録日: 2022-03-07, 公開日: 2022-05-04, 最終更新日: 2023-10-18)

主引用文献

Chang, C.,Lee Luo, C.,Gao, Y.
In crystallo observation of three metal ion promoted DNA polymerase misincorporation.
Nat Commun, 13:2346-2346, 2022

PubMed Abstract: Error-free replication of DNA is essential for life. Despite the proofreading capability of several polymerases, intrinsic polymerase fidelity is in general much higher than what base-pairing energies can provide. Although researchers have investigated this long-standing question with kinetics, structural determination, and computational simulations, the structural factors that dictate polymerase fidelity are not fully resolved. Time-resolved crystallography has elucidated correct nucleotide incorporation and established a three-metal-ion-dependent catalytic mechanism for polymerases. Using X-ray time-resolved crystallography, we visualize the complete DNA misincorporation process catalyzed by DNA polymerase η. The resulting molecular snapshots suggest primer 3´-OH alignment mediated by A-site metal ion binding is the key step in substrate discrimination. Moreover, we observe that C-site metal ion binding preceded the nucleotidyl transfer reaction and demonstrate that the C-site metal ion is strictly required for misincorporation. Our results highlight the essential but separate roles of the three metal ions in DNA synthesis.

PubMed: 35487947
DOI: 10.1038/s41467-022-30005-3

実験手法

X-RAY DIFFRACTION (1.55 Å)