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Structure paper

TitleTwo Distinct Modes of DNA Binding by an MCM Helicase Enable DNA Translocation.
Journal, issue, pagesInt J Mol Sci, Vol. 23, Issue 23, Year 2022
Publish dateNov 24, 2022
AuthorsMartin Meagher / Alexander Myasnikov / Eric J Enemark /
PubMed AbstractA six-subunit ATPase ring forms the central hub of the replication forks in all domains of life. This ring performs a helicase function to separate the two complementary DNA strands to be replicated ...A six-subunit ATPase ring forms the central hub of the replication forks in all domains of life. This ring performs a helicase function to separate the two complementary DNA strands to be replicated and drives the replication machinery along the DNA. Disruption of this helicase/ATPase ring is associated with genetic instability and diseases such as cancer. The helicase/ATPase rings of eukaryotes and archaea consist of six minichromosome maintenance (MCM) proteins. Prior structural studies have shown that MCM rings bind one encircled strand of DNA in a spiral staircase, suggesting that the ring pulls this strand of DNA through its central pore in a hand-over-hand mechanism where the subunit at the bottom of the staircase dissociates from DNA and re-binds DNA one step above the staircase. With high-resolution cryo-EM, we show that the MCM ring of the archaeal organism binds an encircled DNA strand in two different modes with different numbers of subunits engaged to DNA, illustrating a plausible mechanism for the alternating steps of DNA dissociation and re-association that occur during DNA translocation.
External linksInt J Mol Sci / PubMed:36499022 / PubMed Central
MethodsEM (single particle)
Resolution2.34 - 3.01 Å
Structure data

EMDB-27974, PDB-8eaf:
SsoMCM hexamer bound to Mg/ADP-BeFx and 12-mer oligo-dT. Class 1
Method: EM (single particle) / Resolution: 2.62 Å

EMDB-27975, PDB-8eag:
SsoMCM hexamer bound to Mg/ADP-BeFx and 12-mer oligo-dT. Class 2
Method: EM (single particle) / Resolution: 3.01 Å

EMDB-27976, PDB-8eah:
SsoMCM hexamer bound to Mg/ADP-BeFx and 16-mer oligo-dT. Class 1
Method: EM (single particle) / Resolution: 2.48 Å

EMDB-27977, PDB-8eai:
SsoMCM hexamer bound to Mg/ADP-BeFx and 16-mer oligo-dT. Class 2
Method: EM (single particle) / Resolution: 2.76 Å

EMDB-27978, PDB-8eaj:
SsoMCM hexamer bound to Mg/ADP-BeFx and 46-mer DNA strand. Class 1
Method: EM (single particle) / Resolution: 2.45 Å

EMDB-27979, PDB-8eak:
SsoMCM hexamer bound to Mg/ADP-BeFx and 46-mer DNA strand. Class 2
Method: EM (single particle) / Resolution: 2.67 Å

EMDB-27980, PDB-8eal:
SsoMCM hexamer bound to Mg/ADP-BeFx and DNA. Class 1. Merged particles from datasets with 3 different DNA entities
Method: EM (single particle) / Resolution: 2.34 Å

EMDB-27981, PDB-8eam:
SsoMCM hexamer bound to Mg/ADP-BeFx and DNA. Class 2. Merged particles from datasets with 3 different DNA entities
Method: EM (single particle) / Resolution: 2.59 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

ChemComp-08T:
[[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-bis(oxidanyl)oxolan-2-yl]methoxy-oxidanyl-phosphoryl]oxy-oxidanyl-phosphoryl]oxy-tris(fluoranyl)beryllium

ChemComp-HOH:
WATER / Water

Source
  • saccharolobus solfataricus p2 (archaea)
  • synthetic construct (others)
KeywordsREPLICATION / TRANSFERASE/DNA / Helicase / ATPase / TRANSFERASE-DNA complex

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