EMDB/PDB/SASBDBから引用されている文献のデータベース

キーワード
構造解析手法	All X線回折 NMR 電子顕微鏡小角散乱中性子線回折線維回折粉末回折赤外分光 EPR FRET 理論モデルハイブリッド
著者・登録者
ジャーナル
IF	>30 >20 >10 >5 all

タイトル	Structural insights into the exchange mechanism of a replicative DNA polymerase.
ジャーナル・号・ページ	Nucleic Acids Res, Vol. 53, Issue 22, Year 2025
掲載日	2025年11月26日
著者	Xiang Feng / Michelle M Spiering / Almeida Ruda de Luna Santos / Stephen J Benkovic / Huilin Li /
PubMed 要旨	Replicative DNA polymerases are distinguished by their speed, processivity, and fidelity. While speed and fidelity arise from the polymerase's intrinsic catalytic and proofreading activities, ...Replicative DNA polymerases are distinguished by their speed, processivity, and fidelity. While speed and fidelity arise from the polymerase's intrinsic catalytic and proofreading activities, processivity is typically attributed to the DNA sliding clamp that tethers the polymerase to DNA. However, additional mechanisms may also contribute. The T4 bacteriophage polymerase can exchange on-the-fly, a process likely contributing to its ∼10-fold higher synthesis rate compared with human polymerases. Here, we reconstituted the T4 holoenzyme and polymerase exchange complexes using purified gp43 polymerase, gp45 sliding clamp, and a primer-template DNA substrate. Cryo-electron microscopy (cryo-EM) analysis revealed either one or two polymerases bound to the clamp and DNA. In the one-polymerase complex, the DNA threads perpendicularly through the clamp, supporting processive synthesis. In contrast, the two-polymerase complex displays a markedly tilted DNA orientation, impeding sliding and representing exchange intermediates. Three distinct conformational states of the two-polymerase complex define a multistep exchange mechanism. To our knowledge, these findings provide the first molecular-level view of replicative polymerase exchange.
リンク	Nucleic Acids Res / PubMed:41459747 / PubMed Central
手法	EM (単粒子)
解像度	3.57 - 3.99 Å
構造データ	47531 9e5y EMDB-47531: T4 bacteriophage replicative holoenzyme, DNA in right Polymerase(State 1) PDB-9e5y: T4 Bacteriophage Replicative Polymerase Captured in Polymerase Exchange State 1 手法: EM (単粒子) / 解像度: 3.57 Å 47824 9ea2 EMDB-47824: T4 bacteriophage replicative holoenzyme, DNA in left Polymerase(State 2) PDB-9ea2: T4 Bacteriophage Replicative Polymerase Captured in Polymerase Exchange State 2 手法: EM (単粒子) / 解像度: 3.7 Å 47825 9ea3 EMDB-47825: T4 bacteriophage replicative holoenzyme with triple mutants D75R, Q430E, and K432E on exo- Polymerase PDB-9ea3: T4 bacteriophage replicative holoenzyme containing triple mutations D75R, Q430E, and K432E in the exonuclease-deficient polymerase 手法: EM (単粒子) / 解像度: 3.99 Å 47826 9ea6 EMDB-47826: T4 bacteriophage replicative holoenzyme, DNA in left Polymerase(State 3) PDB-9ea6: T4 Bacteriophage Replicative Polymerase Captured in Polymerase Exchange State 3 手法: EM (単粒子) / 解像度: 3.74 Å
化合物	ChemComp-CA: Unknown entry ChemComp-D3T: 2',3'-DIDEOXY-THYMIDINE-5'-TRIPHOSPHATE
由来	escherichia phage t4 (ファージ) synthetic construct (人工物)
キーワード	replication/DNA / replication / T4 / holoenzyme / replication-DNA complex / Transferase/DNA / Transferase-DNA complex