Biomolecular condensates at sites of DNA damage: More than just a phase DOI Creative Commons

Vincent Spegg,

Matthias Altmeyer

DNA repair, Journal Year: 2021, Volume and Issue: 106, P. 103179 - 103179

Published: July 14, 2021

Protein recruitment to DNA break sites is an integral part of the damage response (DDR). Elucidation hierarchy and temporal order with which sensors as well repair signaling factors assemble around chromosome breaks has painted a complex picture tightly regulated macromolecular interactions that build specialized compartments facilitate maintenance genome integrity. While many underlying interactions, e.g. between damage-induced histone marks, can be explained by lock-and-key or induced fit binding models assuming fixed stoichiometries, structurally less defined such highly dynamic multivalent implicated in phase separation, also participate formation multi-protein assemblies genotoxic stress. Although much remains learned about these types cooperative their functional roles, rapidly growing interest material properties biomolecular condensates concepts from polymer chemistry soft matter physics understand biological processes at different scales holds great promises. Here, we discuss nuclear context integrity maintenance, highlighting potential clustered stoichiometric separation. Rather than viewing them opposing scenarios, combined effects balance structural specificity favorable physicochemical relevant for regulation function multilayered condensates.

Language: Английский

Mechanisms of BRCA1–BARD1 nucleosome recognition and ubiquitylation DOI
Qi Hu, Maria Victoria Botuyan, Debiao Zhao

et al.

Nature, Journal Year: 2021, Volume and Issue: 596(7872), P. 438 - 443

Published: July 28, 2021

Language: Английский

Citations

111
RHINO directs MMEJ to repair DNA breaks in mitosis DOI
Alessandra Brambati, Olivia Sacco, Sarina Y. Porcella

et al.

Science, Journal Year: 2023, Volume and Issue: 381(6658), P. 653 - 660

Published: July 13, 2023

Nonhomologous end-joining (NHEJ) and homologous recombination (HR) are the primary pathways for repairing DNA double-strand breaks (DSBs) during interphase, whereas microhomology-mediated (MMEJ) has been regarded as a backup mechanism. Through CRISPR-Cas9-based synthetic lethal screens in cancer cells, we identified subunits of 9-1-1 complex (RAD9A-RAD1-HUS1) its interacting partner, RHINO, crucial MMEJ factors. We uncovered an unexpected function RHINO restricting to mitosis. accumulates M phase, undergoes Polo-like kinase 1 (PLK1) phosphorylation, interacts with polymerase θ (Polθ), enabling recruitment DSBs subsequent repair. Additionally, provide evidence that activity mitosis repairs persistent originate S phase. Our findings offer insights into relationship between genes

Language: Английский

Citations

63
DNA Repair and Therapeutic Strategies in Cancer Stem Cells DOI Open Access

Matthew S. Gillespie,

Ciara Ward,

Clare C. Davies

et al.

Cancers, Journal Year: 2023, Volume and Issue: 15(6), P. 1897 - 1897

Published: March 22, 2023

First-line cancer treatments successfully eradicate the differentiated tumour mass but are comparatively ineffective against stem cells (CSCs), a self-renewing subpopulation thought to be responsible for initiation, metastasis, heterogeneity, and recurrence. CSCs thus presented as principal target elimination during treatment. However, challenging drug because of numerous intrinsic extrinsic mechanisms resistance. One such mechanism that remains relatively understudied is DNA damage response (DDR). presumed possess properties enable enhanced repair efficiency relative their highly proliferative bulk progeny, facilitating improved double-strand breaks induced by radiotherapy most chemotherapeutics. This can occur through multiple mechanisms, including increased expression splicing fidelity genes, robust activation cell cycle checkpoints, elevated homologous recombination-mediated repair. Herein, we summarise current knowledge concerning genome integrity in non-transformed CSCs, discuss therapeutic opportunities within DDR re-sensitising genotoxic stressors, consider challenges posed regarding unbiased identification novel DDR-directed strategies CSCs. A better understanding mediating chemo/radioresistance could lead approaches, thereby enhancing treatment efficacy patients.

Language: Английский

Citations

43
Decoding chromatin states by proteomic profiling of nucleosome readers DOI Creative Commons
Saulius Lukauskas, Andrey Tvardovskiy, Nhuong V. Nguyen

et al.

Nature, Journal Year: 2024, Volume and Issue: 627(8004), P. 671 - 679

Published: March 6, 2024

Abstract DNA and histone modifications combine into characteristic patterns that demarcate functional regions of the genome 1,2 . While many ‘readers’ individual have been described 3–5 , how chromatin states comprising composite modification signatures, variants internucleosomal linker are interpreted is a major open question. Here we use multidimensional proteomics strategy to systematically examine interaction around 2,000 nuclear proteins with over 80 modified dinucleosomes representing promoter, enhancer heterochromatin states. By deconvoluting complex nucleosome-binding profiles networks co-regulated distinct nucleosomal features driving protein recruitment or exclusion, show comprehensively decoded by readers. We find highly distinctive binding responses different features, factors recognize multiple operate largely independently in regulating chromatin. Our online resource, Modification Atlas Regulation Chromatin States (MARCS), provides in-depth analysis tools engage our results advance discovery fundamental principles regulation

Language: Английский

Citations

30
The fork protection complex promotes parental histone recycling and epigenetic memory DOI Creative Commons
Sebastian Jespersen Charlton, Valentin Flury, Yutaka Kanoh

et al.

Cell, Journal Year: 2024, Volume and Issue: 187(18), P. 5029 - 5047.e21

Published: Aug. 1, 2024

The inheritance of parental histones across the replication fork is thought to mediate epigenetic memory. Here, we reveal that fission yeast Mrc1 (CLASPIN in humans) binds H3-H4 tetramers and operates as a central coordinator symmetric histone inheritance. mutants key connector domain disrupted segregation lagging strand comparable Mcm2 histone-binding mutants. Both showed clonal asymmetric loss H3K9me-mediated gene silencing. AlphaFold predicted co-chaperoning by Mcm2, with bridging binding. Biochemical functional analysis validated this model revealed duality function: disabling binding lagging-strand recycling while another mutation impaired leading recycling. We propose toggles between pathways, part intra-replisome co-chaperoning, ensure transmission both daughter cells.

Language: Английский

Citations

19
Dynamic histone modification patterns coordinating DNA processes DOI

Laura López-Hernández,

Patrick Toolan-Kerr, Andrew J. Bannister

et al.

Molecular Cell, Journal Year: 2025, Volume and Issue: 85(2), P. 225 - 237

Published: Jan. 1, 2025

Language: Английский

Citations

2
Nuclear and genome dynamics underlying DNA double-strand break repair DOI
Irene Chiolo, Matthias Altmeyer, Gaëlle Legube

et al.

Nature Reviews Molecular Cell Biology, Journal Year: 2025, Volume and Issue: unknown

Published: March 17, 2025

Language: Английский

Citations

2
Templated Insertions: A Smoking Gun for Polymerase Theta-Mediated End Joining DOI
Joost Schimmel, Robin van Schendel, Johan T. den Dunnen

et al.

Trends in Genetics, Journal Year: 2019, Volume and Issue: 35(9), P. 632 - 644

Published: July 8, 2019

Language: Английский

Citations

127
Studying DNA Double-Strand Break Repair: An Ever-Growing Toolbox DOI Creative Commons
Alexandra C Vítor, Pablo Huertas, Gaëlle Legube

et al.

Frontiers in Molecular Biosciences, Journal Year: 2020, Volume and Issue: 7

Published: Feb. 21, 2020

To ward off against the catastrophic consequences of persistent DNA double-strand breaks (DSBs), eukaryotic cells have developed a set complex signaling networks that detect these lesions, orchestrate cell cycle checkpoints and ultimately lead to their repair. Collectively, comprise damage response (DDR). The current knowledge molecular determinants mechanistic details DDR owes greatly continuous development ground-breaking experimental tools couple controlled induction DSBs at distinct genomic positions with assays reporters investigate repair pathways, impact on other DNA-templated processes specific contribution chromatin environment. In this review, we present tools, discuss pros cons illustrate our understanding DDR. ​

Language: Английский

Citations

127
Stimulation of CRISPR-mediated homology-directed repair by an engineered RAD18 variant DOI Creative Commons

Tarun S. Nambiar,

Pierre Billon,

Giacomo Diedenhofen

et al.

Nature Communications, Journal Year: 2019, Volume and Issue: 10(1)

Published: July 30, 2019

Precise editing of genomic DNA can be achieved upon repair CRISPR-induced double-stranded breaks (DSBs) by homology-directed (HDR). However, the efficiency this process is limited DSB pathways competing with HDR, such as non-homologous end joining (NHEJ). Here we individually express in human cells 204 open reading frames involved damage response (DDR) and determine their impact on CRISPR-mediated HDR. From these studies, identify RAD18 a stimulator By defining domains required to promote derive an enhanced variant (e18) that stimulates HDR multiple cell types, including embryonic stem cells. Mechanistically, e18 induces suppressing localization NHEJ-promoting factor 53BP1 DSBs. Altogether, study identifies enhancer highlights promise engineering DDR factors augment precision genome editing.

Language: Английский

Citations

112