Mechanistic investigation of human maturation of Okazaki fragments reveals slow kinetics.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
15 11 2022
15 11 2022
Historique:
received:
12
06
2022
accepted:
04
11
2022
entrez:
15
11
2022
pubmed:
16
11
2022
medline:
19
11
2022
Statut:
epublish
Résumé
The final steps of lagging strand synthesis induce maturation of Okazaki fragments via removal of the RNA primers and ligation. Iterative cycles between Polymerase δ (Polδ) and Flap endonuclease-1 (FEN1) remove the primer, with an intermediary nick structure generated for each cycle. Here, we show that human Polδ is inefficient in releasing the nick product from FEN1, resulting in non-processive and remarkably slow RNA removal. Ligase 1 (Lig1) can release the nick from FEN1 and actively drive the reaction toward ligation. These mechanisms are coordinated by PCNA, which encircles DNA, and dynamically recruits Polδ, FEN1, and Lig1 to compete for their substrates. Our findings call for investigating additional pathways that may accelerate RNA removal in human cells, such as RNA pre-removal by RNase Hs, which, as demonstrated herein, enhances the maturation rate ~10-fold. They also suggest that FEN1 may attenuate the various activities of Polδ during DNA repair and recombination.
Identifiants
pubmed: 36379932
doi: 10.1038/s41467-022-34751-2
pii: 10.1038/s41467-022-34751-2
pmc: PMC9666535
doi:
Substances chimiques
DNA
9007-49-2
DNA Polymerase III
EC 2.7.7.7
Flap Endonucleases
EC 3.1.-
Okazaki fragments
0
RNA
63231-63-0
FEN1 protein, human
EC 3.1.11.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
6973Informations de copyright
© 2022. The Author(s).
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