A5056528432- DNA Repair Mechanisms
- CRISPR and Genetic Engineering
- T-cell and B-cell Immunology
- Carcinogens and Genotoxicity Assessment
- Ubiquitin and proteasome pathways
- Hematopoietic Stem Cell Transplantation
- CAR-T cell therapy research
- Epigenetics and DNA Methylation
- Immunotherapy and Immune Responses
- Immune Cell Function and Interaction
- Cancer-related Molecular Pathways
- Advanced biosensing and bioanalysis techniques
- PARP inhibition in cancer therapy
- Pluripotent Stem Cells Research
- Cancer therapeutics and mechanisms
- DNA and Nucleic Acid Chemistry
- RNA and protein synthesis mechanisms
- Plant Disease Resistance and Genetics
- Liver physiology and pathology
- Glycosylation and Glycoproteins Research
- Parasitic Infections and Diagnostics
- Polyomavirus and related diseases
- Genetic factors in colorectal cancer
- Genetic Neurodegenerative Diseases
- vaccines and immunoinformatics approaches
The Netherlands Cancer Institute
2011-2024
Oncode Institute
2009-2023
Google (United States)
2008
Genesys (United States)
2005
Leiden University
1996
B cells use translesion DNA synthesis (TLS) to introduce somatic mutations around genetic lesions caused by activation-induced cytidine deaminase. Monoubiquitination at lysine(164) of proliferating cell nuclear antigen (PCNA(K164)) stimulates TLS. To determine the role PCNA(K164) modifications in hypermutation, PCNA(K164R) knock-in mice were generated. PCNA(K164R/K164R) mutants are born a sub-Mendelian frequency. Although proliferate and class switch normally, mutation spectrum hypermutated...
Histone ubiquitination at DNA breaks is required for activation of the damage response (DDR) and repair. How dynamic removal this modification by deubiquitinating enzymes (DUBs) impacts genome maintenance in vivo largely unknown. To address question, we generated mice deficient Ub-specific protease 3 (USP3; Usp3Δ/Δ), a histone H2A DUB which negatively regulates ubiquitin-dependent DDR signaling. Notably, USP3 deletion increased levels adult tissues, reduced hematopoietic stem cell (HSC)...
To determine the feasibility, toxicity, and immunologic effects of vaccination with autologous tumor cells retrovirally transduced GM-CSF gene, we performed a phase I/II study in stage IV metastatic melanoma patients.Sixty-four patients were randomly assigned to receive three vaccinations high-dose or low-dose at 3-week intervals. Tumor cell vaccine preparation succeeded for 56 (88%), but because progressive disease, well-tolerated was completed only 28 patients. We analyzed priming T...
PrimPol is a DNA damage tolerant polymerase displaying both translesion synthesis (TLS) and (re)-priming properties. This led us to study the consequences of deficiency in tolerating mutagenic lesions induced by members APOBEC/AID family cytosine deaminases. Interestingly, during somatic hypermutation, counteracts generation C>G transversions on leading strand. Independently, mutation analyses human invasive breast cancer confirmed pro-mutagenic activity APOBEC3B revealed genome-wide...
Translesion synthesis (TLS) provides a highly conserved mechanism that enables DNA on damaged template. TLS is performed by specialized polymerases of which polymerase (Pol) κ important for the cellular response to damage induced benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE), ultraviolet (UV) light and alkylating agent methyl methanesulfonate (MMS). As are intrinsically error-prone, tight regulation their activity required. One level control provided ubiquitination homotrimeric clamp...
During somatic hypermutation (SHM), B cells introduce mutations into their immunoglobulin genes to generate high affinity antibodies. Current models suggest a separation in the generation of G/C transversions by Ung2-dependent pathway and A/T Msh2/ubiquitinated proliferating cell nuclear antigen (PCNA-Ub)–dependent pathway. It is currently unknown whether these pathways compete initiate mutagenesis PCNA-Ub functions downstream Ung2. Furthermore, do not explain why mice lacking Msh2 have more...
Somatic hypermutation (SHM) and class switch recombination (CSR) of immunoglobulin (Ig) genes are initiated by the enzymatic deamination cytosine (C) to uracil (U). Uracil-DNA-glycosylase (Ung2) converts uracils into apyrimidinic (AP) sites, which is essential for generation transversions (TVs) at G/C basepairs during SHM efficient DNA break formation CSR. Besides Ung2, mismatch repair protein Msh2 translesion synthesis (TLS) polymerase (Pol) Rev1 implicated in To further unravel role Rev1,...
The Aicda locus encodes the activation induced cytidine deaminase (AID) and is highly expressed in germinal center (GC) B cells to initiate somatic hypermutation (SHM) class switch recombination (CSR) of immunoglobulin (Ig) genes. Besides these Ig specific activities cells, AID has been implicated active DNA demethylation non-B cell systems. We here determined a potential role as an epigenetic eraser transcriptional regulator cells. RNA-Seq on different subsets revealed that Aicda−/− are...
Abstract Crosslink repair depends on the Fanconi anemia pathway and translesion synthesis polymerases that replicate over unhooked crosslinks. Translesion is regulated via ubiquitination of PCNA, independently polymerase REV1. The division labor between PCNA-ubiquitination REV1 in interstrand crosslink unclear. Inhibition either these pathways has been proposed as a strategy to increase cytotoxicity platinating agents cancer treatment. Here, we defined importance for DNA mammalian ICL...
DNA damage tolerance (DDT) enables replication to continue in the presence of a damaged template and constitutes key step interstrand crosslink repair. In this way DDT minimizes stress inflicted by wide range endogenous exogenous agents, provides critical first line defense against alkylating platinating chemotherapeutics. Effective strongly depends on damage-induced, site-specific PCNA-ubiquitination at Lysine (K) 164 E2/E3 complex (RAD6/18). A survey The Cancer Genome Atlas (TCGA) revealed...
DNA damage threatens genomic integrity and instigates stem cell failure. To bypass genotoxic lesions during replication, cells employ tolerance (DDT), which is regulated via PCNA ubiquitination REV1. DDT conserved in all domains of life, yet its relevance mammals remains unclear. Here, we show that inactivation both PCNA-ubiquitination REV1 results embryonic adult lethality, the accumulation hematopoietic progenitor (HSPCs) ultimately resulted their depletion. Our reveal crucial maintenance...
The antihelmintic drug ABZ and its metabolites belong to the chemical family of benzimidazoles (BZM) that act as potent tubulin polymerization inhibitors, suggesting a potential re-direction BZMs for cancer therapy. Applying UV-Vis spectrometry we here demonstrate DNA intercalator. This insight led us determine primary mode action in mammalian cells. As revealed by RNA sequencing, did neither grossly affect replication analyzed survival stress signaling, nor transcriptome. Actually, unbiased...
Proliferating cell nuclear antigen (PCNA) encircles DNA as a ring-shaped homotrimer and, by tethering polymerases to their template, PCNA serves critical replication factor. In contrast high-fidelity polymerases, the activation of low-fidelity translesion synthesis (TLS) seems require damage-inducible monoubiquitylation (Ub) at lysine residue 164 (PCNA-Ub). TLS can tolerate damage, i.e. they replicate across lesions. The lack proofreading activity, however, renders highly mutagenic....
The data described here provide genome-wide expression profiles of murine primitive hematopoietic stem and progenitor cells (LSK) B cell populations, obtained by high throughput sequencing. Cells are derived from wild-type mice deficient for the ubiquitin-specific protease 3 (USP3; Usp3Δ/Δ). Modification histone proteins ubiquitin plays a crucial role in cellular response to DNA damage (DDR) (Jackson Durocher, 2013) [1]. USP3 is H2A deubiquitinating enzyme (DUB) that regulates...
The development and differentiation of B cells is intimately linked to cell proliferation the generation diverse immunoglobulin gene (Ig) repertoires. ubiquitin E3 ligase HUWE1 controls proliferation, DNA damage responses, repair, including base excision repair (BER) pathway. These processes are crucial importance for B-cell in bone marrow, germinal center (GC) response, which results clonal expansion expressing high affinity immunoglobulins. Here, we re-examined role Ig diversification,...
Fanconi anemia (FA) develops due to a mutation in one of the FANC genes that are involved repair interstrand crosslinks (ICLs). FANCG, member FA core complex, is essential for ICL repair. Previous FANCG-deficient mouse models were generated with drug-based selection cassettes mixed mice backgrounds, leading disparity interpretation genotype-related phenotype. We created Fancg-KO (KO) model using CRISPR/Cas9 exclude these confounders. The entire Fancg locus was targeted and maintained on...
Monoubiquitylation of the homotrimeric DNA sliding clamp PCNA at lysine residue 164 (PCNAK164) is a highly conserved, damage-inducible process that mediated by E2/E3 complex Rad6/Rad18. This ubiquitylation event recruits translesion synthesis (TLS) polymerases capable replicating across damaged templates. Besides PCNA, Rad6/Rad18 was recently shown in yeast to ubiquitylate also 9-1-1, heterotrimeric composed Rad9, Rad1, and Hus1 manner. Based on similar crystal structures K185 Rad1...
To generate high affinity antibodies during an immune response, B cells undergo somatic hypermutation (SHM) of their immunoglobulin genes. Error-prone translesion synthesis (TLS) DNA polymerases have been reported to be responsible for all mutations at template A/T and least a fraction G/C transversions. In contrast which depend on PCNA ubiquitination, it remains unclear how transversions are regulated SHM. Several lines evidence indicate mechanistic link between the Fanconi Anemia (FA)...
The MET protooncogene, c-MET, encodes a cell surface tyrosine kinase receptor. ligand for c-MET is hepatocyte growth factor (HGF), also known as scatter (SF), which to affect proliferation and motility of primarily epithelial cells. Recently, HGF/SF was shown haemopoiesis. Studies with transfected NIH3T3 cells indicated that the HGF/SF-c-MET interaction promotes invasion in vitro vivo. We previously demonstrated induces adhesion c-MET-positive B-lymphoma extracellular matrix molecules,...
Fanconi anemia (FA) develops due to a mutation in one of the FANC genes that are involved repair interstrand crosslinks (ICLs). FANCG, member FA core complex, is essential for ICL repair. Previous FANCG-deficient mouse models were generated with drug-based selection cassettes mixed mice backgrounds, leading disparity interpretation genotype-related phenotype. To exclude these confounders, we created Fancg-KO (KO) model using CRISPR/Cas9. The entire Fancg locus was targeted and maintained on...
The Fanconi anemia (FA) repair pathway governs of highly genotoxic DNA interstrand crosslinks (ICLs) and relies on translesion synthesis (TLS). TLS is facilitated by REV1 or site-specific monoubiquitination proliferating cell nuclear antigen (PCNA) (PCNA-Ub) at lysine 164 (K164). A
DNA damage tolerance (DDT) enables replication to continue in the presence of fork stalling lesions. In mammalian cells, DDT is regulated by two independent pathways, controlled polymerase REV1 and ubiquitinated PCNA, respectively.