Human DNA polymerase θ harbors DNA end-trimming activity critical for DNA repair.
DNA polymerase
Double-strand break repair
dideoxynucleotides
endonuclease
theta-mediated end joining (TMEJ)
Journal
Molecular cell
ISSN: 1097-4164
Titre abrégé: Mol Cell
Pays: United States
ID NLM: 9802571
Informations de publication
Date de publication:
01 04 2021
01 04 2021
Historique:
received:
19
08
2020
revised:
24
11
2020
accepted:
15
01
2021
pubmed:
13
2
2021
medline:
15
4
2021
entrez:
12
2
2021
Statut:
ppublish
Résumé
Cancers with hereditary defects in homologous recombination rely on DNA polymerase θ (pol θ) for repair of DNA double-strand breaks. During end joining, pol θ aligns microhomology tracts internal to 5'-resected broken ends. An unidentified nuclease trims the 3' ends before synthesis can occur. Here we report that a nuclease activity, which differs from the proofreading activity often associated with DNA polymerases, is intrinsic to the polymerase domain of pol θ. Like the DNA synthesis activity, the nuclease activity requires conserved metal-binding residues, metal ions, and dNTPs and is inhibited by ddNTPs or chain-terminated DNA. Our data indicate that pol θ repurposes metal ions in the polymerase active site for endonucleolytic cleavage and that the polymerase-active and end-trimming conformations of the enzyme are distinct. We reveal a nimble strategy of substrate processing that allows pol θ to trim or extend DNA depending on the DNA repair context.
Identifiants
pubmed: 33577776
pii: S1097-2765(21)00041-1
doi: 10.1016/j.molcel.2021.01.021
pmc: PMC8231307
mid: NIHMS1673585
pii:
doi:
Substances chimiques
Metals
0
DNA
9007-49-2
DNA-Directed DNA Polymerase
EC 2.7.7.7
Endonucleases
EC 3.1.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1534-1547.e4Subventions
Organisme : NCI NIH HHS
ID : P01 CA193124
Pays : United States
Organisme : Howard Hughes Medical Institute
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA052040
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA215990
Pays : United States
Organisme : NCI NIH HHS
ID : P01 CA247773
Pays : United States
Informations de copyright
Copyright © 2021 Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests R.D.W. is a scientific advisor for Repare Therapeutics and a shareholder. R.B.J. is named on patent US9150897B2, which references the phCMV1-2xMBP vector adapted by this study to express pol θ-FL.
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