- EMDB-45400: Cryo-EM structure of a human MCM2-7 double hexamer formed from in... -
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基本情報
登録情報
データベース: EMDB / ID: EMD-45400
タイトル
Cryo-EM structure of a human MCM2-7 double hexamer formed from independently loaded MCM2-7 single hexamers
マップデータ
DeepEMhancer sharpened EM map
試料
複合体: In vitro loaded, human MCM2-7 double hexamer on dsDNA (formed by dimerization of independently loaded MCM2-7 single hexamers)
タンパク質・ペプチド: x 6種
DNA: x 2種
リガンド: x 4種
キーワード
complex / helicase / replication / AAA+ ATPase
機能・相同性
機能・相同性情報
Switching of origins to a post-replicative state / Unwinding of DNA / nuclear origin of replication recognition complex / alpha DNA polymerase:primase complex / Regulation of MITF-M-dependent genes involved in DNA replication, damage repair and senescence / mitotic DNA replication / regulation of phosphorylation / CMG complex / MCM complex / double-strand break repair via break-induced replication ...Switching of origins to a post-replicative state / Unwinding of DNA / nuclear origin of replication recognition complex / alpha DNA polymerase:primase complex / Regulation of MITF-M-dependent genes involved in DNA replication, damage repair and senescence / mitotic DNA replication / regulation of phosphorylation / CMG complex / MCM complex / double-strand break repair via break-induced replication / mitotic DNA replication initiation / regulation of DNA-templated DNA replication initiation / DNA strand elongation involved in DNA replication / DNA replication origin binding / cochlea development / DNA replication initiation / Activation of the pre-replicative complex / Activation of ATR in response to replication stress / cellular response to interleukin-4 / DNA helicase activity / cellular response to epidermal growth factor stimulus / Assembly of the pre-replicative complex / helicase activity / cellular response to xenobiotic stimulus / Orc1 removal from chromatin / nucleosome assembly / single-stranded DNA binding / histone binding / DNA helicase / chromosome, telomeric region / DNA replication / cell population proliferation / cilium / intracellular membrane-bounded organelle / apoptotic process / DNA damage response / chromatin / perinuclear region of cytoplasm / enzyme binding / ATP hydrolysis activity / DNA binding / zinc ion binding / nucleoplasm / ATP binding / identical protein binding / nucleus / membrane / cytoplasm / cytosol 類似検索 - 分子機能
DNA replication licensing factor MCM2-like, winged-helix domain / : / MCM5, C-terminal domain / DNA replication licensing factor MCM7, winged helix / DNA replication licensing factor Mcm5 / MCM4, winged helix domain / DNA replication licensing factor Mcm3 / Mini-chromosome maintenance complex protein 4 / : / MCM3-like, winged helix domain ...DNA replication licensing factor MCM2-like, winged-helix domain / : / MCM5, C-terminal domain / DNA replication licensing factor MCM7, winged helix / DNA replication licensing factor Mcm5 / MCM4, winged helix domain / DNA replication licensing factor Mcm3 / Mini-chromosome maintenance complex protein 4 / : / MCM3-like, winged helix domain / DNA replication licensing factor Mcm6 / DNA replication licensing factor Mcm7 / Mcm6, C-terminal winged-helix domain / MCM6 C-terminal winged-helix domain / DNA replication licensing factor Mcm2 / Mini-chromosome maintenance protein 2 / Mini-chromosome maintenance, conserved site / MCM family signature. / MCM N-terminal domain / MCM N-terminal domain / MCM OB domain / MCM OB domain / Mini-chromosome maintenance protein / MCM, AAA-lid domain / MCM P-loop domain / MCM AAA-lid domain / MCM family domain profile. / minichromosome maintenance proteins / MCM domain / Nucleic acid-binding, OB-fold / ATPases associated with a variety of cellular activities / AAA+ ATPase domain / P-loop containing nucleoside triphosphate hydrolase 類似検索 - ドメイン・相同性
DNA replication licensing factor MCM3 / DNA replication licensing factor MCM4 / DNA replication licensing factor MCM5 / DNA replication licensing factor MCM7 / DNA replication licensing factor MCM2 / DNA replication licensing factor MCM6 類似検索 - 構成要素
生物種
Homo sapiens (ヒト) / Saccharomyces cerevisiae (パン酵母)
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R01-GM141313
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
T32-GM008283
米国
National Institutes of Health/National Cancer Institute (NIH/NCI)
1F31-CA278331-01
米国
National Institutes of Health/National Cancer Institute (NIH/NCI)
S10OD023603
米国
引用
ジャーナル: Nature / 年: 2024 タイトル: Multiple mechanisms for licensing human replication origins. 著者: Ran Yang / Olivia Hunker / Marleigh Wise / Franziska Bleichert / 要旨: Loading of replicative helicases is obligatory for the assembly of DNA replication machineries. The eukaryotic MCM2-7 replicative helicase motor is deposited onto DNA by the origin recognition ...Loading of replicative helicases is obligatory for the assembly of DNA replication machineries. The eukaryotic MCM2-7 replicative helicase motor is deposited onto DNA by the origin recognition complex (ORC) and co-loader proteins as a head-to-head double hexamer to license replication origins. Although extensively studied in budding yeast, the mechanisms of origin licensing in multicellular eukaryotes remain poorly defined. Here we use biochemical reconstitution and electron microscopy to reconstruct the human MCM loading pathway. We find that unlike in yeast, the ORC6 subunit of the ORC is not essential for-but enhances-human MCM loading. Electron microscopy analyses identify several intermediates en route to MCM double hexamer formation in the presence and absence of ORC6, including a DNA-loaded, closed-ring MCM single hexamer intermediate that can mature into a head-to-head double hexamer through multiple mechanisms. ORC6 and ORC3 facilitate the recruitment of the ORC to the dimerization interface of the first hexamer into MCM-ORC (MO) complexes that are distinct from the yeast MO complex and may orient the ORC for second MCM hexamer loading. Additionally, MCM double hexamer formation can proceed through dimerization of independently loaded MCM single hexamers, promoted by a propensity of human MCM2-7 hexamers to self-dimerize. This flexibility in human MCM loading may provide resilience against cellular replication stress, and the reconstitution system will enable studies addressing outstanding questions regarding DNA replication initiation and replication-coupled events in the future.
全体 : In vitro loaded, human MCM2-7 double hexamer on dsDNA (formed by ...
全体
名称: In vitro loaded, human MCM2-7 double hexamer on dsDNA (formed by dimerization of independently loaded MCM2-7 single hexamers)
要素
複合体: In vitro loaded, human MCM2-7 double hexamer on dsDNA (formed by dimerization of independently loaded MCM2-7 single hexamers)
タンパク質・ペプチド: DNA replication licensing factor MCM2
タンパク質・ペプチド: DNA replication licensing factor MCM3
タンパク質・ペプチド: DNA replication licensing factor MCM4
タンパク質・ペプチド: DNA replication licensing factor MCM5
タンパク質・ペプチド: DNA replication licensing factor MCM6
タンパク質・ペプチド: DNA replication licensing factor MCM7
DNA: DNA (44-MER)
DNA: DNA (44-MER)
リガンド: ZINC ION
リガンド: MAGNESIUM ION
リガンド: ADENOSINE-5'-TRIPHOSPHATE
リガンド: ADENOSINE-5'-DIPHOSPHATE
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超分子 #1: In vitro loaded, human MCM2-7 double hexamer on dsDNA (formed by ...
超分子
名称: In vitro loaded, human MCM2-7 double hexamer on dsDNA (formed by dimerization of independently loaded MCM2-7 single hexamers) タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#8