Single-molecule imaging reveals replication fork coupled formation of G-quadruplex structures hinders local replication stress signaling

Guanine-rich DNA sequences occur throughout the human genome and can transiently form G-quadruplex (G4) structures that may obstruct DNA replication, leading to genomic instability. Here, we apply multi-color single-molecule localization microscopy (SMLM) coupled with robust data-mining algorithms t...

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Vydané v:Nature communications Ročník 12; číslo 1; s. 2525 - 14
Hlavní autori: Lee, Wei Ting C., Yin, Yandong, Morten, Michael J., Tonzi, Peter, Gwo, Pam Pam, Odermatt, Diana C., Modesti, Mauro, Cantor, Sharon B., Gari, Kerstin, Huang, Tony T., Rothenberg, Eli
Médium: Journal Article
Jazyk:English
Vydavateľské údaje: London Nature Publishing Group UK 05.05.2021
Nature Publishing Group
Nature Portfolio
Predmet:
13/106
13/89
14/1
14/34
14/63
631/337/151/2353
631/337/2265
631/80/2373/2238
96/33
Algorithms
Animals
Biophysics
Cell Line
Coupling (molecular)
Damage accumulation
Data mining
Deoxyribonucleic acid
DNA
DNA - chemistry
DNA biosynthesis
DNA Damage
DNA helicase
DNA Helicases - metabolism
DNA Replication - physiology
DNA-binding protein
DNA-Binding Proteins
G-Quadruplexes
Gene sequencing
Genomes
Genomic Instability
Guanine
Humanities and Social Sciences
Humans
Life Sciences
Localization
multidisciplinary
Nucleotide sequence
Recombinant Proteins
Replication
Replication forks
Science
Science (multidisciplinary)
Sf9 Cells
Signaling
Single Molecule Imaging - methods
Synthesis
Super-resolution microscopy
Replisome
Single-molecule biophysics
ISSN:2041-1723, 2041-1723
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Shrnutí:Guanine-rich DNA sequences occur throughout the human genome and can transiently form G-quadruplex (G4) structures that may obstruct DNA replication, leading to genomic instability. Here, we apply multi-color single-molecule localization microscopy (SMLM) coupled with robust data-mining algorithms to quantitatively visualize replication fork (RF)-coupled formation and spatial-association of endogenous G4s. Using this data, we investigate the effects of G4s on replisome dynamics and organization. We show that a small fraction of active replication forks spontaneously form G4s at newly unwound DNA immediately behind the MCM helicase and before nascent DNA synthesis. These G4s locally perturb replisome dynamics and organization by reducing DNA synthesis and limiting the binding of the single-strand DNA-binding protein RPA. We find that the resolution of RF-coupled G4s is mediated by an interplay between RPA and the FANCJ helicase. FANCJ deficiency leads to G4 accumulation, DNA damage at G4-associated replication forks, and silencing of the RPA-mediated replication stress response. Our study provides first-hand evidence of the intrinsic, RF-coupled formation of G4 structures, offering unique mechanistic insights into the interference and regulation of stable G4s at replication forks and their effect on RPA-associated fork signaling and genomic instability. In the genome, repetitive guanine-rich sequences have the potential to spontaneously fold into non-canonical DNA secondary structures known as G-quadruplex (G4). Using novel single-molecule imaging approaches, the authors reveal that G4 formation within active replication forks locally perturb replisome dynamics and damage response signaling, which require RPA and FANCJ for regulation.
Bibliografia:ObjectType-Article-1
SourceType-Scholarly Journals-1
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PMCID: PMC8099879
ISSN:2041-1723
2041-1723
DOI:10.1038/s41467-021-22830-9