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Structure paper

TitleDNSN-1 recruits GINS for CMG helicase assembly during DNA replication initiation in .
Journal, issue, pagesScience, Vol. 381, Issue 6664, Page eadi4932, Year 2023
Publish dateSep 22, 2023
AuthorsYisui Xia / Remi Sonneville / Michael Jenkyn-Bedford / Liqin Ji / Constance Alabert / Ye Hong / Joseph T P Yeeles / Karim P M Labib /
PubMed AbstractAssembly of the CMG (CDC-45-MCM-2-7-GINS) helicase is the key regulated step during eukaryotic DNA replication initiation. Until now, it was unclear whether metazoa require additional factors that ...Assembly of the CMG (CDC-45-MCM-2-7-GINS) helicase is the key regulated step during eukaryotic DNA replication initiation. Until now, it was unclear whether metazoa require additional factors that are not present in yeast. In this work, we show that DNSN-1, the ortholog of human DONSON, functions during helicase assembly in a complex with MUS-101/TOPBP1. DNSN-1 is required to recruit the GINS complex to chromatin, and a cryo-electron microscopy structure indicates that DNSN-1 positions GINS on the MCM-2-7 helicase motor (comprising the six MCM-2 to MCM-7 proteins), by direct binding of DNSN-1 to GINS and MCM-3, using interfaces that we show are important for initiation and essential for viability. These findings identify DNSN-1 as a missing link in our understanding of DNA replication initiation, suggesting that initiation defects underlie the human disease syndrome that results from DONSON mutations.
External linksScience / PubMed:37590372 / PubMed Central
MethodsEM (single particle)
Resolution2.64 - 3.75 Å
Structure data

EMDB-17198: Cryo-EM density map of the C. elegans complex CMG helicase/TIM-1/TIPN-1/DNSN-1 dimer on fork DNA (prior to enrichment of particles containing TIM-1/TIPN-1)
Method: EM (single particle) / Resolution: 3.25 Å

EMDB-17199: Cryo-EM density map of the C. elegans complex CMG helicase/TIM-1/TIPN-1/DNSN-1 dimer on fork DNA (Multi-body Refinement DNSN-1 homodimer)
Method: EM (single particle) / Resolution: 3.44 Å

EMDB-17200: Cryo-EM density map of the C. elegans complex CMG helicase/TIM-1/TIPN-1/DNSN-1 dimer on fork DNA (Multi-body Refinement GINS/CDC-45/DNSN-1 N-terminus)
Method: EM (single particle) / Resolution: 2.88 Å

EMDB-17201: Cryo-EM density map of the C. elegans complex CMG helicase/TIM-1/TIPN-1/DNSN-1 dimer on fork DNA (Multi-body Refinement MCM-2-7 AAA+/ssDNA)
Method: EM (single particle) / Resolution: 3.17 Å

EMDB-17202: Cryo-EM density map of the C. elegans complex CMG helicase/TIM-1/TIPN-1/DNSN-1 dimer on fork DNA (Multi-body Refinement MCM-2-7 NTDs)
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-17203: Cryo-EM density map of the C. elegans complex CMG helicase/TIM-1/TIPN-1/DNSN-1 dimer on fork DNA (Multi-body Refinement TIM-1/TIPN-1/MCM-2-7 NTD/dsDNA)
Method: EM (single particle) / Resolution: 2.64 Å

17204

8ouw

EMDB-17204, PDB-8ouw:
Cryo-EM structure of CMG helicase bound to TIM-1/TIPN-1 and homodimeric DNSN-1 on fork DNA (Caenorhabditis elegans)
Method: EM (single particle) / Resolution: 3.75 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

ChemComp-ANP:
PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER / AMP-PNP, energy-carrying molecule analogue*YM

Source
  • caenorhabditis elegans (invertebrata)
  • saccharomyces cerevisiae (brewer's yeast)
KeywordsREPLICATION / DNA Replication / DNA Replication Initiation / Replisome / DONSON

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