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TitleCryo-EM structures and biochemical insights into heterotrimeric PCNA regulation of DNA ligase.
Journal, issue, pagesStructure, Vol. 30, Issue 3, Page 371-385.e5, Year 2022
Publish dateMar 3, 2022
AuthorsAleksandr Sverzhinsky / Alan E Tomkinson / John M Pascal /
PubMed AbstractDNA ligases act in the final step of many DNA repair pathways and are commonly regulated by the DNA sliding clamp proliferating cell nuclear antigen (PCNA), but there are limited insights into the ...DNA ligases act in the final step of many DNA repair pathways and are commonly regulated by the DNA sliding clamp proliferating cell nuclear antigen (PCNA), but there are limited insights into the physical basis for this regulation. Here, we use single-particle cryoelectron microscopy (cryo-EM) to analyze an archaeal DNA ligase and heterotrimeric PCNA in complex with a single-strand DNA break. The cryo-EM structures highlight a continuous DNA-binding surface formed between DNA ligase and PCNA that supports the distorted conformation of the DNA break undergoing repair and contributes to PCNA stimulation of DNA ligation. DNA ligase is conformationally flexible within the complex, with its domains fully ordered only when encircling the repaired DNA to form a stacked ring structure with PCNA. The structures highlight DNA ligase structural transitions while docked on PCNA, changes in DNA conformation during ligation, and the potential for DNA ligase domains to regulate PCNA accessibility to other repair factors.
External linksStructure / PubMed:34838188 / PubMed Central
MethodsEM (single particle)
Resolution4.16 - 4.38 Å
Structure data

24618

7rpo

EMDB-24618, PDB-7rpo:
Archaeal DNA ligase and heterotrimeric PCNA in complex with non-ligatable DNA
Method: EM (single particle) / Resolution: 4.16 Å

24624

7rpw

EMDB-24624, PDB-7rpw:
Archaeal DNA ligase and heterotrimeric PCNA in complex with adenylated DNA
Method: EM (single particle) / Resolution: 4.38 Å

24625

7rpx

EMDB-24625, PDB-7rpx:
Archaeal DNA ligase and heterotrimeric PCNA in complex with end-joined DNA
Method: EM (single particle) / Resolution: 4.2 Å

Chemicals

ChemComp-MN:
Unknown entry

ChemComp-AMP:
ADENOSINE MONOPHOSPHATE / AMP*YM

Source
  • saccharolobus solfataricus (archaea)
  • homo sapiens (human)
KeywordsLIGASE/DNA / DNA ligase / PCNA / cryo-EM / LIGASE-DNA complex

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