Database of articles cited by EMDB/PDB/SASBDB data

Keywords
Structure methods	All X-ray diffraction NMR electron microscopy small angle scattering neutron diffraction fiber diffraction powder diffraction infrared spectroscopy EPR fluorescence transfer theoretical model Hybrid
Author
Journal
IF	>30 >20 >10 >5 all

Title	MCM double hexamer loading visualized with human proteins.
Journal, issue, pages	Nature, Vol. 636, Issue 8042, Page 499-508, Year 2024
Publish date	Nov 27, 2024
Authors	Florian Weissmann / Julia F Greiwe / Thomas Pühringer / Evelyn L Eastwood / Emma C Couves / Thomas C R Miller / John F X Diffley / Alessandro Costa /
PubMed Abstract	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.
External links	Nature / PubMed:39604733 / PubMed Central
Methods	EM (single particle)
Resolution	3.1 - 5.6 Å
Structure data	19618 8s09 EMDB-19618, PDB-8s09: H. sapiens MCM2-7 double hexamer bound to double stranded DNA Method: EM (single particle) / Resolution: 3.1 Å 19619 8s0a EMDB-19619, PDB-8s0a: H. sapiens MCM2-7 hexamer bound to double stranded DNA Method: EM (single particle) / Resolution: 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 Method: EM (single particle) / Resolution: 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 Method: EM (single particle) / Resolution: 4.0 Å 19622 8s0d EMDB-19622, PDB-8s0d: H. sapiens MCM bound to double stranded DNA and ORC1-6 Method: EM (single particle) / Resolution: 3.6 Å 19623 8s0e EMDB-19623, PDB-8s0e: H. sapiens OCCM bound to double stranded DNA Method: EM (single particle) / Resolution: 3.8 Å 19624 8s0f EMDB-19624, PDB-8s0f: H. sapiens OC1M bound to double stranded DNA Method: EM (single particle) / Resolution: 4.1 Å EMDB-19625: H. sapiens MCM double hexamer containing the MCM5 R195A/L209G mutant Method: EM (single particle) / Resolution: 5.6 Å
Chemicals	ChemComp-ATP: ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying moleculeYM ChemComp-MG: Unknown entry ChemComp-ZN: Unknown entry ChemComp-ADP: ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying moleculeYM ChemComp-AGS: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / ATP-gamma-S, energy-carrying molecule analogue*YM
Source	synthetic construct (others) homo sapiens (human) synthetic construct
Keywords	REPLICATION / AAA+ ATPase / DNA helicase