Conditional knockout of RAD51-related genes in Leishmania major reveals a critical role for homologous recombination during genome replication DOI Creative Commons
Jeziel D. Damasceno, João Luís Reis-Cunha, Kathryn Crouch

et al.

PLoS Genetics, Journal Year: 2020, Volume and Issue: 16(7), P. e1008828 - e1008828

Published: July 1, 2020

Homologous recombination (HR) has an intimate relationship with genome replication, both during repair of DNA lesions that might prevent synthesis and in tackling stalls to the replication fork. Recent studies led us ask if HR have a more central role replicating Leishmania, eukaryotic parasite. Conflicting evidence emerged regarding whether or not genes are essential, genome-wide mapping provided for unorthodox organisation initiation sites, termed origins. To answer this question, we employed combined CRISPR/Cas9 DiCre approach rapidly generate assess effect conditional ablation RAD51 three RAD51-related proteins Leishmania major. Using approach, demonstrate loss any these factors is immediately lethal but each case growth slows time leads damage accumulation cells aberrant content. Despite similarities, show only RAD51-3 impairs causes elevated levels mutation. Furthermore, two act distinct ways, since RAD51, RAD51-3, profound on causing at major origins increased subtelomeres. Our work clarifies questions importance survival reveals unanticipated, programme microbial eukaryote.

Language: Английский

Computed structures of core eukaryotic protein complexes DOI
Ian R. Humphreys, Jimin Pei, Minkyung Baek

et al.

Science, Journal Year: 2021, Volume and Issue: 374(6573)

Published: Nov. 11, 2021

Protein-protein interactions play critical roles in biology, but the structures of many eukaryotic protein complexes are unknown, and there likely not yet identified. We take advantage advances proteome-wide amino acid coevolution analysis deep-learning–based structure modeling to systematically identify build accurate models core within

Language: Английский

Citations

452
The Unfolded Protein Response: Detecting and Responding to Fluctuations in the Protein-Folding Capacity of the Endoplasmic Reticulum DOI Open Access
G Elif Karagöz, Diego Acosta‐Alvear, Peter Walter

et al.

Cold Spring Harbor Perspectives in Biology, Journal Year: 2019, Volume and Issue: 11(9), P. a033886 - a033886

Published: Jan. 22, 2019

Most of the secreted and plasma membrane proteins are synthesized on membrane-bound ribosomes endoplasmic reticulum (ER). They require engagement ER-resident chaperones foldases that assist in their folding maturation. Since protein homeostasis ER is crucial for cellular function, protein-folding status organelle's lumen continually surveyed by a network signaling pathways, collectively called unfolded response (UPR). Protein-folding imbalances, or "ER stress," detected highly conserved sensors adjust ER's capacity according to physiological needs cell. We review recent developments field have provided new insights into stress-sensing mechanisms used UPR which they integrate various inputs organelle accommodate fluctuations demands.

Language: Английский

Citations

263
RAD51 Gene Family Structure and Function DOI Open Access
Braulio Bonilla, Sarah R Hengel, McKenzie K. Grundy

et al.

Annual Review of Genetics, Journal Year: 2020, Volume and Issue: 54(1), P. 25 - 46

Published: July 14, 2020

Accurate DNA repair and replication are critical for genomic stability cancer prevention. RAD51 its gene family key regulators of fidelity through diverse roles in double-strand break repair, stress, meiosis. is an ATPase that forms a nucleoprotein filament on single-stranded DNA. has the function finding invading homologous sequences to enable accurate timely repair. Its paralogs, which arose from ancient duplications RAD51, have evolved regulate promote function. Underscoring importance, misregulation associated with diseases such as Fanconi anemia. In this review, we focus mammalian structure highlight use model systems mechanistic understanding cellular roles. We also discuss how members contributes disease consider new approaches pharmacologically inhibit RAD51.

Language: Английский

Citations

167
Homologous Recombination Deficiency: Cancer Predispositions and Treatment Implications DOI Open Access

Ming-Ren Toh,

Joanne Ngeow

The Oncologist, Journal Year: 2021, Volume and Issue: 26(9), P. e1526 - e1537

Published: May 22, 2021

Abstract Homologous recombination (HR) is a highly accurate DNA repair mechanism. Several HR genes are established cancer susceptibility with clinically actionable pathogenic variants (PVs). Classically, BRCA1 and BRCA2 germline PVs associated significant breast ovarian risks. Patients or display worse clinical outcomes but respond better to platinum-based chemotherapies poly-ADP ribose polymerase inhibitors, trait termed “BRCAness.” With the advent of whole-exome sequencing multigene panels, in other increasingly identified among familial cancers. As such, several such as PALB2 reclassified predisposition genes. But evidence for risks remains unclear many others. In this review, we will discuss predispositions treatment implications beyond BRCA2, focus on 24 genes: 53BP1, ATM, ATR, ATRIP, BARD1, BLM, BRIP1, DMC1, MRE11A, NBN, PALB2, RAD50, RAD51, RAD51B, RAD51C, RAD51D, RIF1, RMI1, RMI2, RPA1, TOP3A, TOPBP1, XRCC2, XRCC3. Implications Practice This review provides comprehensive reference readers quickly identify potential predisposing homologous genes, generate research questions inconclusive evidence. also evaluates “BRCAness” each member. Clinicians can refer these discussions candidates future trials.

Language: Английский

Citations

88
Role of Rad51 and DNA repair in cancer: A molecular perspective DOI
Erik Laurini, Domenico Marson,

Alice Fermeglia

et al.

Pharmacology & Therapeutics, Journal Year: 2020, Volume and Issue: 208, P. 107492 - 107492

Published: Jan. 27, 2020

Language: Английский

Citations

87
Sequential role of RAD51 paralog complexes in replication fork remodeling and restart DOI Creative Commons

Matteo Berti,

Federico Teloni, Sofija Mijic

et al.

Nature Communications, Journal Year: 2020, Volume and Issue: 11(1)

Published: July 15, 2020

Abstract Homologous recombination (HR) factors were recently implicated in DNA replication fork remodeling and protection. While maintaining genome stability, HR-mediated promotes cancer chemoresistance, by as-yet elusive mechanisms. Five HR cofactors – the RAD51 paralogs RAD51B, RAD51C, RAD51D, XRCC2 XRCC3 emerged as crucial tumor suppressors. Albeit extensively characterized repair, their role has not been addressed systematically. Here, we identify all while screening for modulators of recombinase upon stress. Single-molecule analysis progression architecture isogenic cellular systems shows that BCDX2 subcomplex restrains stress, promoting reversal. Accordingly, primes unscheduled degradation reversed forks BRCA2-defective cells, boosting genomic instability. Conversely, CX3 is dispensable reversal, but mediates efficient restart forks. We propose sequentially orchestrate clinically relevant transactions at forks, cooperatively restart.

Language: Английский

Citations

75
High-content imaging-based pooled CRISPR screens in mammalian cells DOI Creative Commons
Xiaowei Yan, Nico Stuurman, Susana A. Ribeiro

et al.

The Journal of Cell Biology, Journal Year: 2021, Volume and Issue: 220(2)

Published: Jan. 19, 2021

CRISPR (clustered regularly interspaced short palindromic repeats)-based gene inactivation provides a powerful means for linking genes to particular cellular phenotypes. CRISPR-based screening typically uses large genomic pools of single guide RNAs (sgRNAs). However, this approach is limited phenotypes that can be enriched by chemical selection or FACS sorting. Here, we developed microscopy-based approach, which name optical enrichment, select cells displaying CRISPR-induced phenotype automated imaging-based computation, mark them photoactivation an expressed photoactivatable fluorescent protein, and then isolate the using fluorescence-activated cell sorting (FACS). A plugin was open source software μManager automate phenotypic identification cells, allowing ∼1.5 million individual screened in 8 h. We used screen 6,092 sgRNAs targeting 544 their effects on nuclear size regulation identified 14 bona fide hits. These results present scalable facilitate pooled screens.

Language: Английский

Citations

71
The Rad51 paralog complex Rad55-Rad57 acts as a molecular chaperone during homologous recombination DOI Creative Commons
Upasana Roy, Youngho Kwon, Léa Marie

et al.

Molecular Cell, Journal Year: 2021, Volume and Issue: 81(5), P. 1043 - 1057.e8

Published: Jan. 12, 2021

Language: Английский

Citations

58
Structural insights into BCDX2 complex function in homologous recombination DOI
Yashpal Rawal,

Lijia Jia,

Aviv Meir

et al.

Nature, Journal Year: 2023, Volume and Issue: 619(7970), P. 640 - 649

Published: June 21, 2023

Language: Английский

Citations

23
Human Fallopian Tube-Derived Organoids with TP53 and RAD51D Mutations Recapitulate an Early Stage High-Grade Serous Ovarian Cancer Phenotype In Vitro DOI Open Access
Yilin Dai, Jing Xu,

Xiaofeng Gong

et al.

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(2), P. 886 - 886

Published: Jan. 10, 2024

RAD51D mutations have been implicated in the transformation of normal fallopian tube epithelial (FTE) cells into high-grade serous ovarian cancer (HGSOC), one most prevalent and aggressive gynecologic malignancies. Currently, no suitable model exists to elucidate role disease initiation progression. Here, we established organoids from primary human FTE introduced TP53 as well knockdown enable exploration their mutational impact on lesion generation. We observed that deletion rescued adverse effects proliferation, stemness, senescence, apoptosis organoids. impaired homologous recombination (HR) function induced G2/M phase arrest, whereas concurrent mitigated G0/G1 arrest boosted DNA replication when combined with mutation. The co-deletion downregulated cilia assembly, development, motility, but upregulated multiple HGSOC-associated pathways, including IL-17 signaling pathway. IL-17A treatment significantly improved cell viability. co-deleted exhibited heightened sensitivity platinum, poly-ADP ribose polymerase inhibitors (PARPi), cycle-related medication. In summary, our research highlighted use an invaluable vitro platform for early detection carcinogenesis, mechanistic exploration, drug screening.

Language: Английский

Citations

9