A5028157175- DNA Repair Mechanisms
- Genomics and Chromatin Dynamics
- CRISPR and Genetic Engineering
- RNA Research and Splicing
- Cancer-related Molecular Pathways
- Ubiquitin and proteasome pathways
- Microtubule and mitosis dynamics
- PARP inhibition in cancer therapy
- Fungal and yeast genetics research
- Cancer therapeutics and mechanisms
- RNA modifications and cancer
- Genetics and Neurodevelopmental Disorders
- Lung Cancer Research Studies
- Microbial Natural Products and Biosynthesis
- RNA Interference and Gene Delivery
- Plant Gene Expression Analysis
- Pneumocystis jirovecii pneumonia detection and treatment
- Plant Virus Research Studies
- Chromatin Remodeling and Cancer
- NF-κB Signaling Pathways
- Neutropenia and Cancer Infections
- Epigenetics and DNA Methylation
- RNA and protein synthesis mechanisms
- Cell death mechanisms and regulation
- Silk-based biomaterials and applications
University of Cambridge
2011-2025
Cancer Research UK
2023-2025
The Gurdon Institute
2010-2023
Wellcome Trust
2010-2020
Howard Hughes Medical Institute
2003-2004
Wayne State University
2003
Rutgers, The State University of New Jersey
2003
The Wistar Institute
1999-2002
The FACT (facilitates chromatin transcription) complex is required for transcript elongation through nucleosomes by RNA polymerase II (Pol II) in vitro. Here, we show that facilitates Pol II-driven transcription destabilizing nucleosomal structure so one histone H2A-H2B dimer removed during enzyme passage. We also demonstrate possesses intrinsic chaperone activity and can deposit core histones onto DNA. Importantly, requires both of its constituent subunits dependent on the highly acidic C...
Chromosomal deletions and rearrangements in tumors are often associated with common fragile sites, which specific genomic loci prone to gaps breaks metaphase chromosomes. Common sites appear arise through incomplete DNA replication because they induced after partial inhibition by agents such as aphidicolin. Here, we show that G1 cells, large nuclear bodies contain p53 binding protein 1 (53BP1), phosphorylated H2AX (γH2AX), mediator of damage checkpoint (MDC1), well components previously...
Protein ubiquitylation and sumoylation play key roles in regulating cellular responses to DNA double-strand breaks (DSBs). Here, we show that human RNF4, a small ubiquitin-like modifier (SUMO)-targeted ubiquitin E3 ligase, is recruited DSBs manner requiring its SUMO interaction motifs, the ligases PIAS1 PIAS4, various DSB-responsive proteins. Furthermore, reveal RNF4 depletion impairs adduct formation at DSB sites causes persistent histone H2AX phosphorylation (γH2AX) associated with...
The signaling cascade initiated in response to DNA double-strand breaks (DSBs) has been extensively investigated interphase cells. Here, we show that mitotic cells treated with DSB-inducing agents activate a “primary” damage (DDR) comprised of early events, including activation the protein kinases ataxia telangiectasia mutated (ATM) and DNA-dependent kinase (DNA-PK), histone H2AX phosphorylation together recruitment mediator checkpoint 1 (MDC1), Mre11–Rad50–Nbs1 (MRN) complex sites. However,...
SAGA, a recently described protein complex in Saccharomyces cerevisiae, is important for transcription vivo and possesses histone acetylation function. Here we report both biochemical genetic analyses of members three classes regulatory factors contained within the SAGA complex. We demonstrate correlation between phenotypic severity mutants structural integrity. Specifically, null mutations Gcn5/Ada2/Ada3 or Spt3/Spt8 cause moderate phenotypes subtle alterations, while third subgroup,...
Abstract ATR plays key roles in cellular responses to DNA damage and replication stress, a pervasive feature of cancer cells. inhibitors (ATRi) are clinical development for treating various cancers, including those with high such as is elicited by ARID1A deficiency, but the mechanisms that determine ATRi efficacy backgrounds unclear. Here, we have conducted unbiased genome-scale CRISPR screens -deficient proficient cells treated ATRi. We found loss transcription factor KLF5 has severe...
Loss of functional BRCA1 protein leads to defects in DNA double-strand break (DSB) repair by homologous recombination (HR) and renders cells hypersensitive poly (ADP-ribose) polymerase (PARP) inhibitors used treat BRCA1/2-deficient cancers. However, upon chronic treatment BRCA1-mutant with PARP inhibitors, resistant clones can arise via several mechanisms, including loss 53BP1 or its downstream co-factors. Defects the axis partially restore ability a BRCA1-deficient cell form RAD51 filaments...
SAGA is a 1.8-MDa yeast protein complex that composed of several distinct classes transcription-related factors, including the adaptor/acetyltransferase Gcn5, Spt proteins, and subset TBP-associated factors. Our results indicate mutations completely disrupt (deletions SPT7 orSPT20) strongly reduce transcriptional activation at theHIS3 TRP3 genes Gcn5 required for normal HIS3 start site selection. Surprisingly, in proteins involved SAGA-TBP interaction (Spt3 Spt8) cause derepression andTRP3...
Spt-Ada-Gcn5 acetyltransferase (SAGA) is a previously described histone acetyltransferase/transcriptional coactivator complex in yeast. At promoters of certain genes ( HIS3 and TRP3 ), SAGA has an inhibitory function involving nonproductive TATA-binding protein interaction mediated by the Spt3 Spt8 subunits. Related to this, Spt8-less major form under activating conditions for these genes. In present study, we purify this activation-specific complex, called SALSA (SAGA altered, absent)....
The DNA damage response (DDR) is critical for genome stability and the suppression of a wide variety human malignancies, including neurodevelopmental disorders, immunodeficiency, cancer. In addition, efficacy many chemotherapeutic strategies dictated by status DDR. Ubiquitin-specific protease 28 (USP28) was reported to govern multiple factors that are diverse aspects Here, we examined effects USP28 depletion on DDR in cells vivo. We found recruited double-strand breaks manner requires tandem...
Abstract Histone H2AX and MDC1 are key DNA repair DNA-damage signalling proteins. When double-strand breaks (DSBs) occur, is phosphorylated then recruits MDC1, which in turn serves as a docking platform to promote the localization of other factors, including 53BP1, DSB sites. Here, by using CRISPR-Cas9 engineered human cell lines, we identify hitherto unknown, H2AX-independent, function mediated its PST-repeat region. We show that region directly interacts with chromatin via nucleosome...
The catalytic cycle of topoisomerase 2 (TOP2) enzymes proceeds via a transient DNA double-strand break (DSB) intermediate termed the TOP2 cleavage complex (TOP2cc), in which protein is covalently bound to DNA. Anticancer agents such as etoposide operate by stabilizing TOP2ccs, ultimately generating genotoxic TOP2-DNA cross-links that require processing and repair. Here, we identify RAD54 like (RAD54L2) factor promoting TOP2cc resolution. We demonstrate RAD54L2 acts through novel mechanism...
The Fanconi anemia pathway orchestrates the repair of DNA interstrand cross-links and stalled replication forks. A key step in this is UBE2T FANCL-dependent monoubiquitylation FANCD2-FANCI complex. represents an attractive therapeutic target, because activation has been linked to chemotherapy resistance several cancers. However, date, very few selective inhibitors ubiquitin conjugation pathways are known. By using a high-throughput screen-compatible assay, we have identified small-molecule...
HAP1 is a near-haploid human cell line commonly used for mutagenesis and genome editing studies due to its hemizygous nature. We noticed an unusual hypersensitivity of camptothecin, antineoplastic drug that stabilizes topoisomerase I cleavage complexes (TOP1ccs). have attributed this deficiency TDP1, key phosphodiesterase involved in resolving abortive TOP1ccs. Through whole-exome sequencing subsequent restoration TDP1 protein via CRISPR-Cas9 endogenous editing, we demonstrate camptothecin...
Histone H2AX and MDC1 are key DNA repair DNA-damage signalling proteins. When double-strand breaks (DSBs) occur, is phosphorylated then recruits MDC1, which in turn serves as a docking platform to promote the localization of other factors, including 53BP1, DSB sites. Here, by using CRISPR-Cas9 engineered human cell lines, we identify hitherto unknown, H2AX-independent, function mediated its PST-repeat region. We show that region directly interacts with chromatin via nucleosome acidic patch...
Loss of functional BRCA1 protein leads to defects in DNA double-strand break (DSB) repair by homologous recombination (HR) and renders cells hypersensitive poly (ADP-ribose) polymerase (PARP) inhibitors used treat BRCA1/2-deficient cancers. However, upon chronic treatment BRCA1-mutant with PARP inhibitors, resistant clones can arise via several mechanisms, including loss 53BP1 or its downstream co-factors. Defects the axis partially restore ability a BRCA1-deficient cell form RAD51 filaments...