The mechanism of replication stalling and recovery within repetitive DNA.
Date
2022-07-19Author
Casas-Delucchi, CS
Daza-Martin, M
Williams, SL
Coster, G
Type
Journal Article
Metadata
Show full item recordAbstract
Accurate chromosomal DNA replication is essential to maintain genomic stability. Genetic evidence suggests that certain repetitive sequences impair replication, yet the underlying mechanism is poorly defined. Replication could be directly inhibited by the DNA template or indirectly, for example by DNA-bound proteins. Here, we reconstitute replication of mono-, di- and trinucleotide repeats in vitro using eukaryotic replisomes assembled from purified proteins. We find that structure-prone repeats are sufficient to impair replication. Whilst template unwinding is unaffected, leading strand synthesis is inhibited, leading to fork uncoupling. Synthesis through hairpin-forming repeats is rescued by replisome-intrinsic mechanisms, whereas synthesis of quadruplex-forming repeats requires an extrinsic accessory helicase. DNA-induced fork stalling is mechanistically similar to that induced by leading strand DNA lesions, highlighting structure-prone repeats as an important potential source of replication stress. Thus, we propose that our understanding of the cellular response to replication stress may also be applied to DNA-induced replication stalling.
Collections
Subject
DNA
DNA Helicases
DNA Replication
Genomic Instability
Humans
Trinucleotide Repeats
Research team
Genome Replication
Language
eng
Date accepted
2022-06-27
License start date
2022-07-19
Citation
Nature Communications, 2022, 13 (1), pp. 3953 -
Publisher
NATURE PORTFOLIO