BibTex format
@article{Wells:2025:10.1038/s41467-024-55772-z,
author = {Wells, J and Edwardes, L and Leber, V and Allyjaun, S and Peach, M and Tomkins, J and Kefala-Stavridi, A and Faull, S and Aramayo, R and Pestana, C and Ranjha, L and Speck, C},
doi = {10.1038/s41467-024-55772-z},
journal = {Nature Communications},
title = {Reconstitution of human DNA licensing and the structural and functional analysis of key intermediates},
url = {http://dx.doi.org/10.1038/s41467-024-55772-z},
volume = {16},
year = {2025}
}
RIS format (EndNote, RefMan)
TY - JOUR
AB - Human DNA licensing initiates replication fork assembly and DNA replication. This reaction promotes the loading of the hMCM2-7 complex on DNA, which represents the core of the replicative helicase that unwinds DNA during S-phase. Here, we report the reconstitution of human DNA licensing using purified proteins. We showed that the in vitro reaction is specific and results in the assembly of high-salt resistant hMCM2-7 double-hexamers. With ATPγS, an hORC1-5-hCDC6-hCDT1-hMCM2-7 (hOCCM) assembles independent of hORC6, but hORC6 enhances double-hexamer formation. We determined the hOCCM structure, which showed that hORC-hCDC6 recruits hMCM2-7 via five hMCM winged-helix domains. The structure highlights how hORC1 activates the hCDC6 ATPase and uncovered an unexpected role for hCDC6 ATPase in complex disassembly. We identified that hCDC6 binding to hORC1-5 stabilises hORC2-DNA interactions and supports hMCM3-dependent recruitment of hMCM2-7. Finally, the structure allowed us to locate cancer-associated mutations at the hCDC6-hMCM3 interface, which showed specific helicase loading defects.
AU - Wells,J
AU - Edwardes,L
AU - Leber,V
AU - Allyjaun,S
AU - Peach,M
AU - Tomkins,J
AU - Kefala-Stavridi,A
AU - Faull,S
AU - Aramayo,R
AU - Pestana,C
AU - Ranjha,L
AU - Speck,C
DO - 10.1038/s41467-024-55772-z
PY - 2025///
SN - 2041-1723
TI - Reconstitution of human DNA licensing and the structural and functional analysis of key intermediates
T2 - Nature Communications
UR - http://dx.doi.org/10.1038/s41467-024-55772-z
UR - https://www.nature.com/articles/s41467-024-55772-z
VL - 16
ER -