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Structure paper

TitleStructure of the PCNA unloader Elg1-RFC.
Journal, issue, pagesSci Adv, Vol. 10, Issue 9, Page eadl1739, Year 2024
Publish dateMar 1, 2024
AuthorsFengwei Zheng / Nina Y Yao / Roxana E Georgescu / Huilin Li / Michael E O'Donnell /
PubMed AbstractDuring DNA replication, the proliferating cell nuclear antigen (PCNA) clamps are loaded onto primed sites for each Okazaki fragment synthesis by the AAA heteropentamer replication factor C (RFC). ...During DNA replication, the proliferating cell nuclear antigen (PCNA) clamps are loaded onto primed sites for each Okazaki fragment synthesis by the AAA heteropentamer replication factor C (RFC). PCNA encircling duplex DNA is quite stable and is removed from DNA by the dedicated clamp unloader Elg1-RFC. Here, we show the cryo-EM structure of Elg1-RFC in various states with PCNA. The structures reveal essential features of Elg1-RFC that explain how it is dedicated to PCNA unloading. Specifically, Elg1 contains two external loops that block opening of the Elg1-RFC complex for DNA binding, and an "Elg1 plug" domain that fills the central DNA binding chamber, thereby reinforcing the exclusive PCNA unloading activity of Elg1-RFC. Elg1-RFC was capable of unloading PCNA using non-hydrolyzable AMP-PNP. Both RFC and Elg1-RFC could remove PCNA from covalently closed circular DNA, indicating that PCNA unloading occurs by a mechanism that is distinct from PCNA loading. Implications for the PCNA unloading mechanism are discussed.
External linksSci Adv / PubMed:38427736 / PubMed Central
MethodsEM (single particle)
Resolution3.2 - 3.67 Å
Structure data

41252

8thb

EMDB-41252: Cryo-EM map of the Saccharomyces cerevisiae PCNA clamp unloader Elg1-RFC complex
PDB-8thb: Structure of the Saccharomyces cerevisiae PCNA clamp unloader Elg1-RFC complex
Method: EM (single particle) / Resolution: 3.2 Å

41253

8thc

EMDB-41253: Cryo-EM map of the Saccharomyces cerevisiae clamp unloader Elg1-RFC bound to a cracked PCNA
PDB-8thc: Structure of the Saccharomyces cerevisiae clamp unloader Elg1-RFC bound to a cracked PCNA
Method: EM (single particle) / Resolution: 3.67 Å

41254

8thd

EMDB-41254: Cryo-EM map of the Saccharomyces cerevisiae clamp unloader Elg1-RFC bound to PCNA
PDB-8thd: Structure of the Saccharomyces cerevisiae clamp unloader Elg1-RFC bound to PCNA
Method: EM (single particle) / Resolution: 3.25 Å

Chemicals

ChemComp-AGS:
PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / ATP-gamma-S, energy-carrying molecule analogue*YM

ChemComp-MG:
Unknown entry

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

Source
  • saccharomyces cerevisiae (brewer's yeast)
KeywordsREPLICATION / DNA replication / DNA sliding clamp / PCNA clamp / clamp unloader / Elg1-RFC unloader / clamp loader/unloader

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