Dsk1 repairs DSBs by homologous recombination via Rad52 in Schizosaccharomyces pombe
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1099302 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR28637411_1.fastq.gz | Fastqsanger.gz | |||
| SRR28637411_2.fastq.gz | Fastqsanger.gz | |||
| SRR28637412_1.fastq.gz | Fastqsanger.gz | |||
| SRR28637412_2.fastq.gz | Fastqsanger.gz | |||
| SRR28637413_1.fastq.gz | Fastqsanger.gz |
Items per page: 5 1 - 5 of 12 |
Similar Datasets
Project description:Homologous recombination (HR) repairs double-stranded DNA breaks (DSBs). The DSBs are resected to yield single-stranded DNA (ssDNA) that are coated by Replication Protein A (RPA). Rad51 is a recombinase and catalyzes strand invasion and the search for homology. However, it binds to ssDNA with lower affinity than RPA. Thus, mediator proteins such as Rad52 (yeast) or BRCA2 (humans) are required to promote Rad51 binding to RPA-coated ssDNA, but the underlying mechanisms remain poorly understood. Saccharomyces cerevisiae Rad52 interacts with Rad51 through two distinct binding modes. We here uncover that the Rad51-binding site in the disordered C-terminus of Rad52 sorts polydisperse Rad51 into discrete monomers. We developed fluorescent-Rad51 to directly visualize filament formation in single molecule optical tweezer analysis and capture Rad52 catalyzed Rad51 loading onto RPA-coated ssDNA with a distinct preference for junctions. Addition of the Rad51-paralog Rad55-Rad57 increases the number of Rad51 bound by ~60%. Deletion of the C-terminus of Rad52 results in loss of Rad51 sorting and abrogates Rad51 binding to RPA-coated DNA. While BRCA2 and Rad52 are structurally unrelated, we show that many of the functional features are conserved. We describe a concerted Sort, Stack & Extend (SSE) mechanism for mediator-paralog catalyzed Rad51 filament formation in HR.
2025-07-28 | PXD065573 | Pride
Project description:To investigate the different of gene expression between dsk1Δ and wild strain, and then, we performed the gene expression analysis from the RNA-Seq data.
2024-12-25 | GSE263826 | GEO
Project description:The SWI/SNF family of ATP-dependent chromatin remodeling complexes are implicated in multiple DNA damage response mechanisms and frequently mutated in cancer. The BAF and PBAF complexes are two major types of SWI/SNF complexes that are functionally distinguished by their exclusive subunits. Accumulating evidence suggests that double-strand breaks (DSBs) in transcriptionally active DNA are preferentially repaired by a dedicated homologous recombination pathway. We show that different BAF and PBAF subunits promote homologous recombination and are rapidly recruited to DSBs in a transcription-dependent manner. The PBAF complex promotes RNA polymerase II eviction near DNA damage to rapidly initiate transcriptional silencing, while the BAF complex helps to maintain this transcriptional silencing. Furthermore, ARID1A-containing BAF complexes promote RNaseH1 and RAD52 recruitment to facilitate R-loop resolution and DNA repair. Our results highlight how multiple SWI/SNF complexes perform different functions to enable DNA repair in the context of actively transcribed genes.
2023-07-31 | PXD035140 | Pride
Project description:In homologous recombination deficient cells, polymerase theta (Pol θ)-mediated end-joining (TMEJ) repairs replicative DNA breaks during mitosis through interactions with RHINO and TOPBP1. V(D)J recombination and class switch recombination (CSR) rely on DNA breakage in G1-phase B lymphocytes and repair by non-homologous end-joining (NHEJ). In this study, we assayed CSR in primary B cells deficient in XRCC4, SHLD1, and Pol θ. We found that B cells deficient in both XRCC4 and Pol θ, or SHLD1 and Pol θ, exhibit an increased rate of IgH breaks, while retaining CSR levels similar to those of B cells deficient in XRCC4 or SHLD1. We show that in XRCC4- or SHLD1-deficient B cells, Pol θ promotes the formation of aberrant recombination products characterized by exacerbated double-strand break (DSB) end resection, inversion and microhomology, and that these end-joining events occur independently of RHINO. Furthermore, we find that TMEJ of RAG-generated DSBs is also independent of RHINO and concomitant to S/G2 entry. Based on these results, we propose that in the absence of NHEJ, TMEJ repairs persistent G1-phase DSBs in S/G2, rather than in mitosis.
2025-08-26 | GSE306291 | GEO
Project description:SUMOylation, a conserved post-translational modification in eukaryotes, regulates protein function, localization, and stability. One of the least understood aspects of SUMOylation is formation of polymeric chains and their impact on DNA metabolism. Here, using Schizosaccharomyces pombe, we show that loss of SUMO chains results in elevated spontaneous replication stress and DNA damage. Notably, SUMO chains are required for proper centromeric organization; their absence leads to aberrant recruitment of recombination factors and increased recombination within centromeres. To explore site-specific SUMOylation connected with recombination repair, we established a split-SUMO-ID proteomics approach that allowed identification of SUMO- and Rad52-dependent interactomes. This revealed an enrichment of polySUMOylated proteins at the sites of Rad52 repair, including the essential replication factor PCNA. We demonstrate that SUMO chain-modified PCNA counteracts Rad8-mediated PCNA polyubiquitination, thereby regulating the channeling toward the template-switch branch of post-replication repair at stalled forks within centromeres. In the absence of SUMO chains, excessive template-switching drives elevated recombination at centromeres. Finally, we show that artificial tethering of a SUMO chain to Rad52 suppresses centromeric recombination caused by loss of SUMO chains. Together, our findings uncover an essential role for SUMO chains in maintaining centromere stability by modulating DNA repair pathway choice under endogenous replication stress.
2024-09-15 | GSE276805 | GEO
Project description:We report the development of a novel strand-specific ChIP-seq strategy and application of this strategy in studying genome instability events We examined vulnerable genomic sites by applying strand-specific ChIP-seq of Rad52 to S. pombe haploid mutants showing elevated level of genome instability.
2012-07-07 | E-GEOD-39166 | biostudies-arrayexpress
Project description:The level of H2BK5acK10acK15ac affects the repair of DSBs in Schizosaccharomyces pombe
| PRJNA1103096 | ENA
Project description:Precise double-strand break (DSB) signaling and repair is paramount for maintaining genome stability. Homologous recombination (HR) is the chosen DSB repair pathway when cyclin-dependent kinase (CDK) activity is high, as it correlates well with the availability of an intact sister chromatid to be used as a template. However, the late stages of mitosis, anaphase and telophase, are paradoxical scenarios since high CDK levels favor HR repair despite sister chromatids being no longer aligned. To identify factors that specifically are involved in DSB repair in late mitosis, we have now undertaken a comparative proteomic analysis in Saccharomyces cerevisiae and found that Msc1, a poorly characterized protein previously identified as important in meiotic HR, is significantly enriched upon both random and guided DSBs. We further show that the knockout mutant for MSC1 is more sensitive to DSBs in late mitosis, and that msc1Δ has a delayed repair of DBSs as indicated by increased Rad53 hyperphosphorylation, fewer Rad52 repair factories and slower HR completion. We have found that Msc1 is an NE protein that faces the NE lumen and tends to form patches in nuclear halves that contain Rad52 factories. Either depletion or overexpression of Msc1 leads to DSB-independent abnormal nuclear morphologies in late mitosis, including blebbing, compartmentalization and premature signs of karyokinesis. In this regard, one of the two Msc1 orthologs in Schizosaccharomyces pombe, Les1, has been shown to regulate karyokinesis. We discuss how Msc1 may protect the late NE from abnormal events during DSB repair, providing a previously unreported link between NE homeostasis and DSB repair in late mitosis.
2024-08-10 | PXD043515 | Pride
Project description:Gene disruption by CRISPR/Cas9 is highly efficient and relies on the error-prone non-homologous end-joining (NHEJ) pathway. Conversely, precise gene editing requires homology-directed repair (HDR), which occurs at a lower frequency than NHEJ in mammalian cells. Here, by testing whether manipulation of DNA repair factors would improve HDR efficacy, we show that transient ectopic co-expression of RAD52 and a dominant-negative 53BP1 (dn53BP1) synergize to enable efficient HDR using a single-stranded oligonucleotide DNA donor template at multiple loci in human cells, including patient-derived induced pluripotent stem (iPS) cells. Co-expression of RAD52 and dn53BP1 improves multiplexed HDR-mediated editing, whereas expression of RAD52 alone enhances HDR with Cas9 nickase. Our data show that the frequency of NHEJ-mediated DSB repair in the presence of these two factors is not suppressed, and suggest that dn53BP1 competitively antagonizes 53BP1 to augment HDR in combination with RAD52. Importantly, co-expression of RAD52 and dn53BP1 does not alter Cas9 off-target activity. These findings support the use of RAD52 and dn53BP1 co-expression to overcome bottlenecks that limit HDR in precision genome editing.
2017-11-23 | GSE103241 | GEO
Project description:Serine-Arginine Protein Kinases (SRPKs) play important role in diverse biological processes such as alternative splicing and cell cycle. However, the functions and mechanisms of SRPKs in DNA damage response and repair remain unclear. Here we characterized the DNA repair function of SRPKs homolog Dsk1 in the fission yeast Schizosaccharomyces pombe.
2024-07-09 | PXD053831 |