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TitleMechanism of human Lig1 regulation by PCNA in Okazaki fragment sealing.
Journal, issue, pagesNat Commun, Vol. 13, Issue 1, Page 7833, Year 2022
Publish dateDec 20, 2022
AuthorsKerry Blair / Muhammad Tehseen / Vlad-Stefan Raducanu / Taha Shahid / Claudia Lancey / Fahad Rashid / Ramon Crehuet / Samir M Hamdan / Alfredo De Biasio /
PubMed AbstractDuring lagging strand synthesis, DNA Ligase 1 (Lig1) cooperates with the sliding clamp PCNA to seal the nicks between Okazaki fragments generated by Pol δ and Flap endonuclease 1 (FEN1). We present ...During lagging strand synthesis, DNA Ligase 1 (Lig1) cooperates with the sliding clamp PCNA to seal the nicks between Okazaki fragments generated by Pol δ and Flap endonuclease 1 (FEN1). We present several cryo-EM structures combined with functional assays, showing that human Lig1 recruits PCNA to nicked DNA using two PCNA-interacting motifs (PIPs) located at its disordered N-terminus (PIP) and DNA binding domain (PIP). Once Lig1 and PCNA assemble as two-stack rings encircling DNA, PIP is released from PCNA and only PIP is required for ligation to facilitate the substrate handoff from FEN1. Consistently, we observed that PCNA forms a defined complex with FEN1 and nicked DNA, and it recruits Lig1 to an unoccupied monomer creating a toolbelt that drives the transfer of DNA to Lig1. Collectively, our results provide a structural model on how PCNA regulates FEN1 and Lig1 during Okazaki fragments maturation.
External linksNat Commun / PubMed:36539424 / PubMed Central
MethodsEM (single particle)
Resolution4.2 - 7.8 Å
Structure data

14078

7qnz

EMDB-14078, PDB-7qnz:
human Lig1-DNA-PCNA complex reconstituted in absence of ATP
Method: EM (single particle) / Resolution: 4.58 Å

14080

7qo1

EMDB-14080, PDB-7qo1:
complex of DNA ligase I and FEN1 on PCNA and DNA
Method: EM (single particle) / Resolution: 4.4 Å

EMDB-15385: FEN1 holoenzyme with a nicked DNA substrate
Method: EM (single particle) / Resolution: 7.8 Å

15921

8b8t

EMDB-15921, PDB-8b8t:
Open conformation of the complex of DNA ligase I on PCNA and DNA in the presence of ATP
Method: EM (single particle) / Resolution: 4.2 Å

Chemicals

ChemComp-AMP:
ADENOSINE MONOPHOSPHATE / AMP*YM

Source
  • homo sapiens (human)
  • synthetic construct (others)
KeywordsREPLICATION / DNA / Complex / Ligase / PCNA / Ligation / Okazaki fragment maturation / FEN1 / Flap Endonuclease I

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