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	MCM double hexamer loading visualized with human proteins.
ジャーナル・号・ページ	Nature, Vol. 636, Issue 8042, Page 499-508, Year 2024
掲載日	2024年11月27日
著者	Florian Weissmann / Julia F Greiwe / Thomas Pühringer / Evelyn L Eastwood / Emma C Couves / Thomas C R Miller / John F X Diffley / Alessandro Costa /
PubMed 要旨	Eukaryotic DNA replication begins with the loading of the MCM replicative DNA helicase as a head-to-head double hexamer at origins of DNA replication. Our current understanding of how the double ...Eukaryotic DNA replication begins with the loading of the MCM replicative DNA helicase as a head-to-head double hexamer at origins of DNA replication. Our current understanding of how the double hexamer is assembled by the origin recognition complex (ORC), CDC6 and CDT1 comes mostly from budding yeast. Here we characterize human double hexamer (hDH) loading using biochemical reconstitution and cryo-electron microscopy with purified proteins. We show that the human double hexamer engages DNA differently from the yeast double hexamer (yDH), and generates approximately five base pairs of underwound DNA at the interface between hexamers, as seen in hDH isolated from cells. We identify several differences from the yeast double hexamer in the order of factor recruitment and dependencies during hDH assembly. Unlike in yeast, the ORC6 subunit of the ORC is not essential for initial MCM recruitment or hDH loading, but contributes to an alternative hDH assembly pathway that requires an intrinsically disordered region in ORC1, which may work through a MCM-ORC intermediate. Our work presents a detailed view of how double hexamers are assembled in an organism that uses sequence-independent replication origins, provides further evidence for diversity in eukaryotic double hexamer assembly mechanisms, and represents a first step towards reconstitution of DNA replication initiation with purified human proteins.
リンク	Nature / PubMed:39604733 / PubMed Central
手法	EM (単粒子)
解像度	3.1 - 5.6 Å
構造データ	19618 8s09 EMDB-19618, PDB-8s09: H. sapiens MCM2-7 double hexamer bound to double stranded DNA 手法: EM (単粒子) / 解像度: 3.1 Å 19619 8s0a EMDB-19619, PDB-8s0a: H. sapiens MCM2-7 hexamer bound to double stranded DNA 手法: EM (単粒子) / 解像度: 3.2 Å 19620 8s0b EMDB-19620, PDB-8s0b: H. sapiens MCM bound to double stranded DNA and ORC6 as part of the MCM-ORC complex 手法: EM (単粒子) / 解像度: 3.6 Å 19621 8s0c EMDB-19621, PDB-8s0c: H. sapiens ORC1-5 bound to double stranded DNA as part of the MCM-ORC complex 手法: EM (単粒子) / 解像度: 4.0 Å 19622 8s0d EMDB-19622, PDB-8s0d: H. sapiens MCM bound to double stranded DNA and ORC1-6 手法: EM (単粒子) / 解像度: 3.6 Å 19623 8s0e EMDB-19623, PDB-8s0e: H. sapiens OCCM bound to double stranded DNA 手法: EM (単粒子) / 解像度: 3.8 Å 19624 8s0f EMDB-19624, PDB-8s0f: H. sapiens OC1M bound to double stranded DNA 手法: EM (単粒子) / 解像度: 4.1 Å EMDB-19625: H. sapiens MCM double hexamer containing the MCM5 R195A/L209G mutant 手法: EM (単粒子) / 解像度: 5.6 Å
化合物	ChemComp-ATP: ADENOSINE-5'-TRIPHOSPHATE / ATP / ATP, エネルギー貯蔵分子YM ChemComp-MG: Unknown entry / マグネシウムジカチオン ChemComp-ZN: Unknown entry ChemComp-ADP: ADENOSINE-5'-DIPHOSPHATE / ADP / ADP, エネルギー貯蔵分子YM ChemComp-AGS: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / ATP-γ-S / ATP-gamma-S, エネルギー貯蔵分子類似体*YM
由来	synthetic construct (人工物) homo sapiens (ヒト) synthetic construct
キーワード	REPLICATION / AAA+ ATPase / DNA helicase