A5014568357- DNA Repair Mechanisms
- Genomics and Chromatin Dynamics
- CRISPR and Genetic Engineering
- RNA Research and Splicing
- Genetics and Neurodevelopmental Disorders
- Carcinogens and Genotoxicity Assessment
- RNA modifications and cancer
- RNA regulation and disease
- Ubiquitin and proteasome pathways
- DNA and Nucleic Acid Chemistry
- PARP inhibition in cancer therapy
- RNA and protein synthesis mechanisms
- Advanced biosensing and bioanalysis techniques
- Hereditary Neurological Disorders
- Porphyrin Metabolism and Disorders
- NF-κB Signaling Pathways
- Chromatin Remodeling and Cancer
- Cancer-related Molecular Pathways
- Mitochondrial Function and Pathology
- Genetically Modified Organisms Research
- Skin and Cellular Biology Research
- Nuclear Structure and Function
- Glycosylation and Glycoproteins Research
- RNA Interference and Gene Delivery
- Fungal and yeast genetics research
Erasmus MC
2013-2024
Erasmus MC Cancer Institute
2021-2024
Oncode Institute
2018-2022
Cancer Genomics Centre
2014-2017
Erasmus University Rotterdam
2009-2015
University of Oslo
2012
Institut de Pharmacologie et de Biologie Structurale
2012
Oslo University Hospital
2012
Centre National de la Recherche Scientifique
2012
Medical Genetics Center
2006
AbstractInterstrand cross-links (ICLs) are an extremely toxic class of DNA damage incurred during normal metabolism or cancer chemotherapy. ICLs covalently tether both strands duplex DNA, preventing the strand unwinding that is essential for polymerase access. The mechanism ICL repair in mammalian cells poorly understood. However, genetic data implicate Ercc1-Xpf endonuclease and proteins required homologous recombination-mediated double-strand break (DSB) repair. To examine role repair, we...
The DNA polymerase processivity factor proliferating cell nuclear antigen (PCNA) is central to both replication and repair. ring-shaped homotrimeric PCNA encircles slides along double-stranded DNA, acting as a "sliding clamp" that localizes proteins DNA. We determined the behavior of green fluorescent protein-tagged human (GFP-hPCNA) in living cells analyze its different engagements Photobleaching tracking foci revealed dynamic equilibrium between two kinetic pools PCNA, i.e., bound free...
Significance Transcription by RNA Polymerase II (Pol II) is a highly dynamic process that tightly regulated at each step of the transcription cycle. We generated GFP-RPB1 knockin cells and developed photobleaching endogenous Pol combined with computational modeling to study in vivo dynamics real time. This approach allowed us dissect promoter-paused from initiating elongating showed initiation promoter proximal pausing are surprisingly events, due premature termination II. Our provides new...
XPC recognizes UV-induced DNA lesions and initiates their removal by nucleotide excision repair (NER). Damage recognition in NER is tightly controlled ubiquitin SUMO modifications. Recent studies have shown that the SUMO-targeted ligase RNF111 promotes K63-linked ubiquitylation of SUMOylated after damage. However, exact regulatory function these modifications vivo remains elusive. Here we show required for efficient ultraviolet-induced lesions. RNF111-mediated release from damaged...
Sensitivity and resistance of cells to platinum drug chemotherapy are a large extent determined by activity the DNA damage response (DDR). Combining with inhibition specific DDR pathways could therefore improve treatment efficacy. Multiple have been implicated in removal platinum-DNA lesions, but it is unclear which exact most important cellular resistance. Here, we used CRISPR/Cas9 screening identify proteins that protect colorectal cancer against clinically applied oxaliplatin. We find...
UV-DDB, consisting of subunits DDB1 and DDB2, recognizes UV-induced photoproducts during global genome nucleotide excision repair (GG-NER). We recently demonstrated a noncanonical role UV-DDB in stimulating base (BER) which raised several questions about the timing arrival at 8-oxoguanine (8-oxoG), dependency on recruitment downstream BER NER proteins. Using two different approaches to introduce 8-oxoG cells, we show that DDB2 is recruited immediately after damage colocalizes with...
Abstract The precise regulation of RNA Polymerase II (Pol II) transcription after genotoxic stress is crucial for proper execution the DNA damage-induced response. While stalling Pol on transcription-blocking lesions (TBLs) blocks transcript elongation and initiates repair in cis, TBLs additionally elicit a response trans that regulates genome-wide. Here we uncover that, an initial block trigger genome-wide VCP-mediated proteasomal degradation promoter-bound, P-Ser5-modified trans. This...
Abstract The SWI/SNF family of ATP-dependent chromatin remodeling complexes is implicated in multiple DNA damage response mechanisms and frequently mutated cancer. BAF, PBAF ncBAF are three major types that functionally distinguished by their exclusive subunits. Accumulating evidence suggests double-strand breaks (DSBs) transcriptionally active preferentially repaired a dedicated homologous recombination pathway. We show different subunits promote rapidly recruited to DSBs...
Transcription-coupled nucleotide excision repair factor Cockayne syndrome protein B (CSB) was suggested to function in the of oxidative DNA damage. However thus far, no clear role for CSB base (BER), dedicated pathway remove abundant damage, could be established. Using live cell imaging with a laser-assisted procedure locally induce 8-oxo-7,8-dihydroguanine (8-oxoG) lesions, we previously showed that is recruited these lesions transcription-dependent but NER-independent fashion. Here...
Abstract DNA damage sensors DDB2 and XPC initiate global genome nucleotide excision repair (NER) to protect from mutagenesis caused by helix-distorting lesions. recognizes helical distortions binding unpaired ssDNA opposite binds UV-induced lesions directly facilitates efficient recognition XPC. We show that not only lesion-binding but also timely dissociation is required for handover swift progression of the multistep reaction. DNA-binding-induced ubiquitylation ensuing degradation regulate...
Transcription/repair factor IIH (TFIIH) is essential for RNA polymerase II transcription and nucleotide excision repair (NER). This multi-subunit complex consists of ten polypeptides, including the recently identified small 8-kDa trichothiodystrophy group A (TTDA)/ hTFB5 protein. Patients belonging to rare neurodevelopmental syndrome TTD-A carry inactivating mutations in TTDA/hTFB5 gene. One these completely inactivates protein, whereas other TFIIH genes only tolerate point that do not...
Xeroderma pigmentosum (XP) is caused by defects in the nucleotide excision repair (NER) pathway. NER removes helix-distorting DNA lesions, such as UV–induced photodimers, from genome. Patients suffering XP exhibit exquisite sun sensitivity, high incidence of skin cancer, and some cases neurodegeneration. The severity varies tremendously depending upon which gene mutated how severely mutation affects capacity. XPF-ERCC1 a structure-specific endonuclease essential for incising damaged strand...
Abstract Congenital nucleotide excision repair (NER) deficiency gives rise to several cancer-prone and/or progeroid disorders. It is not understood how defects in the same DNA pathway cause different disease features and severity. Here, we show that absence of functional ERCC1-XPF or XPG endonucleases leads stable prolonged binding transcription/DNA factor TFIIH damage, which correlates with severity induces senescence human cells. In vivo, C. elegans , this non-excised damage causes...
The rare recessive developmental disorder Trichothiodystrophy (TTD) is characterized by brittle hair and nails. Patients also present a variable set of poorly explained additional clinical features, including ichthyosis, impaired intelligence, delay anemia. About half TTD patients are photosensitive due to inherited defects in the DNA repair transcription factor II H (TFIIH). pathophysiological contributions unrepaired lesions have not been dissected yet. Here, we functionally characterize...
Trichothiodystrophy (TTD) is a rare hereditary neurodevelopmental disorder defined by sulfur-deficient brittle hair and nails scaly skin, but with otherwise remarkably variable clinical features. The photosensitive TTD (PS-TTD) forms exhibits in addition to progressive neuropathy other features of segmental accelerated aging associated impaired genome maintenance transcription. New factors involved various steps gene expression have been identified for the different non-photosensitive...
The position of chromosomal neighborhoods in living cells was followed using three different methods for marking domains occupying arbitrary locations the nucleus; photobleaching GFP-labeled histone H2B, local UV-marked DNA, and fluorescently labeled DNA. All revealed that global organization can be reestablished through one cell division from mother to daughters. By simultaneously monitoring cycle stage which relative domain positions were tracked, we observed neighborhood is apparently...