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Title | Structural Insight into the MCM double hexamer activation by Dbf4-Cdc7 kinase. |
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Journal, issue, pages | Nat Commun, Vol. 13, Issue 1, Page 1396, Year 2022 |
Publish date | Mar 16, 2022 |
Authors | Jiaxuan Cheng / Ningning Li / Yunjing Huo / Shangyu Dang / Bik-Kwoon Tye / Ning Gao / Yuanliang Zhai / |
PubMed Abstract | The Dbf4-dependent kinase Cdc7 (DDK) regulates DNA replication initiation by phosphorylation of the MCM double hexamer (MCM-DH) to promote helicase activation. Here, we determine a series of cryo ...The Dbf4-dependent kinase Cdc7 (DDK) regulates DNA replication initiation by phosphorylation of the MCM double hexamer (MCM-DH) to promote helicase activation. Here, we determine a series of cryo electron microscopy (cryo-EM) structures of yeast DDK bound to the MCM-DH. These structures, occupied by one or two DDKs, differ primarily in the conformations of the kinase core. The interactions of DDK with the MCM-DH are mediated exclusively by subunit Dbf4 straddling across the hexamer interface on the three N-terminal domains (NTDs) of subunits Mcm2, Mcm6, and Mcm4. This arrangement brings Cdc7 close to its only essential substrate, the N-terminal serine/threonine-rich domain (NSD) of Mcm4. Dbf4 further displaces the NSD from its binding site on Mcm4-NTD, facilitating an immediate targeting of this motif by Cdc7. Moreover, the active center of Cdc7 is occupied by a unique Dbf4 inhibitory loop, which is disengaged when the kinase core assumes wobbling conformations. This study elucidates the versatility of Dbf4 in regulating the ordered multisite phosphorylation of the MCM-DH by Cdc7 kinase during helicase activation. |
External links | Nat Commun / PubMed:35296675 / PubMed Central |
Methods | EM (single particle) |
Resolution | 2.52 - 4.0 Å |
Structure data | EMDB-31684, PDB-7v3u: EMDB-31685, PDB-7v3v: EMDB-31686: Cryo-EM map of DDK subtracted from DH-DDK complex EMDB-31687: Cryo-EM map of Dbf4-NTD engaged with Mcm2-NTD-A subtracted from DH-DDK complex EMDB-31688: Cryo-EM structure of MCM double hexamer bound with two DDKs (Group I) EMDB-31689: Cryo-EM structure of MCM double hexamer bound with two DDKs (Group II) EMDB-31690: Cryo-EM structure of MCM double hexamer bound with two DDKs (Group III) EMDB-31691: Cryo-EM structure of MCM double hexamer bound with one DDK (Group IV) EMDB-31692: Cryo-EM structure of MCM double hexamer bound with one DDK (Group V) EMDB-31694: Cryo-EM structure of mutant MCM double hexamer (Mcm4 delta N140) bound with DDK EMDB-31695: Cryo-EM structure of mutant MCM double hexamer (Mcm4 delta N174) bound with DDK EMDB-31696: Cryo-EM structure of MCM double hexamer bound with DDK in State II EMDB-31699: Cryo-EM structure of mutant MCM double hexamer (Mcm4 delta N140) EMDB-31700: Cryo-EM structure of mutant MCM double hexamer (Mcm4 delta N174) EMDB-31701: Cryo-EM structure of MCM double hexamer phosphorylated by DDK EMDB-32355, PDB-7w8g: |
Chemicals | ChemComp-AGS: ChemComp-MG: ChemComp-ZN: ChemComp-ADP: |
Source |
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Keywords | CELL CYCLE / DNA replication initiation / Complex / Replicative helicase / Replication / Kinase |