A5003390908- DNA Repair Mechanisms
- Genomics and Chromatin Dynamics
- PARP inhibition in cancer therapy
- CRISPR and Genetic Engineering
- Microtubule and mitosis dynamics
- Integrated Circuits and Semiconductor Failure Analysis
- Cancer-related Molecular Pathways
- Epigenetics and DNA Methylation
- Cellular transport and secretion
- Toxin Mechanisms and Immunotoxins
- RNA Research and Splicing
- Calcium signaling and nucleotide metabolism
- Bacterial Genetics and Biotechnology
- Mitochondrial Function and Pathology
- Pluripotent Stem Cells Research
- Biosimilars and Bioanalytical Methods
- Chromosomal and Genetic Variations
- Sirtuins and Resveratrol in Medicine
- Nuclear Structure and Function
- RNA and protein synthesis mechanisms
University of Zurich
2015-2021
Life Science Zurich
2016-2021
University of Michigan
2016
Abstract Intrinsically disordered proteins can phase separate from the soluble intracellular space, and tend to aggregate under pathological conditions. The physiological functions molecular triggers of liquid demixing by separation are not well understood. Here we show in vitro vivo that nucleic acid-mimicking biopolymer poly(ADP-ribose) (PAR) nucleates demixing. PAR levels markedly induced at sites DNA damage, provide evidence PAR-seeded results rapid, yet transient fully reversible...
Embryonic stem cells (ESCs) represent a transient biological state, where pluripotency is coupled with fast proliferation. ESCs display constitutively active DNA damage response (DDR), but its molecular determinants have remained elusive. Here we show in cultured and mouse embryos that H2AX phosphorylation dependent on Ataxia telangiectasia Rad3 related (ATR) associated chromatin loading of the ssDNA-binding proteins RPA RAD51. Single-molecule analysis replication intermediates reveals...
The bivalent histone modification reader 53BP1 accumulates around DNA double-strand breaks (DSBs), where it dictates repair pathway choice decisions by limiting end resection. How this function is regulated locally and across the cell cycle to channel reactions toward non-homologous joining (NHEJ) in G1 promote homology-directed (HDR) S/G2 insufficiently understood. Here, we show that ability of accumulate DSBs declines as cells progress through S phase reveal inverse relationship between...
Abstract Exploiting the full potential of anti-cancer drugs necessitates a detailed understanding their cytotoxic effects. While standard omics approaches are limited to cell population averages, emerging single techniques currently lack throughput and not applicable for compound screens. Here, we employed versatile sensitive high-content microscopy-based approach overcome these limitations quantify multiple parameters cytotoxicity at level in cycle resolved manner. Applied PARP inhibitors...
Cellular mechanisms that safeguard genome integrity are often subverted in cancer. To identify cancer-related caretakers, we employed a convergent multi-screening strategy coupled to quantitative image-based cytometry and ranked candidate genes according multivariate readouts reflecting viability, proliferative capacity, replisome integrity, DNA damage signaling. This unveiled regulators of replication stress resilience, including components the pre-mRNA cleavage polyadenylation complex. We...
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...
In addition to its role as an electron transporter, mitochondrial nicotinamide adenine dinucleotide (NAD+) is important co-factor for enzymatic reactions, including ADP-ribosylation. Although mitochondria harbor the most intra-cellular NAD+, ADP-ribosylation remains poorly understood. Here we provide evidence ADP-ribosylation, which was identified using various methodologies immunofluorescence, western blot, and mass spectrometry. We show that reversibly increases in response respiratory...
The p53-Mdm2 system is key to tumor suppression. We have recently reported that p53 as well Mdm2 are capable of supporting DNA replication fork progression. On the other hand, we found a modifier chromatin, modulating polycomb repressor complex (PRC)-driven histone modifications. Here show that, similar knockdown, depletion PRC members impairs synthesis, determined in fiber assays. In particular, ubiquitin ligase and PRC1 component RNF2/Ring1B required support replication, Mdm2. Moreover,...
The DNA replication machinery frequently encounters impediments that slow fork progression and threaten timely error-free replication. CHK1 protein kinase is essential to deal with stress (RS) ensure genome integrity cell survival, yet how basal levels activity of are maintained under physiological, unstressed conditions not well understood. Here, we reveal stability controlled by its steady-state during unchallenged proliferation. This autoactivatory mechanism, which depends on ATR...
Accurate repair of DNA double-strand breaks (DSBs) is essential for genome stability, and defective underlies diseases such as cancer. Homologous recombination uses an intact homologous sequence to faithfully restore damaged DNA, yet how broken ends find sites in a containing billions non-homologous bases remains unclear. Here, we introduce sister-pore-C, high-resolution method mapping intra- trans-molecular interactions replicated chromosomes. We show that DSBs reshape chromosome...
Abstract The human genome encodes 45 kinesin motor proteins that drive cell division, motility, intracellular trafficking and ciliary function. Determining the cellular function of each would benefit from specific small-molecule inhibitors. However, screens have yielded only a few Here we present novel chemical-genetic approach to engineer motors can carry out wild-type yet also be efficiently inhibited by small, cell-permeable molecules. Using kinesin-1 as prototype, develop two independent...
The iron-sulfur (FeS) cluster helicase DDX11 is associated with a human disorder termed Warsaw Breakage Syndrome. Interestingly, one disease-associated mutation affects the highly conserved arginine-263 in FeS cluster-binding motif. Here, we demonstrate that required for DNA binding, ATP hydrolysis, and activity, most likely because of its positive charge. We further show interacts replication factors polymerase delta WDHD1. In vitro, can remove obstacles ahead Pol δ an ATPase-...
Harmful oxidation of proteins, lipids and nucleic acids is observed when reactive oxygen species (ROS) are produced excessively and/or the antioxidant capacity reduced, causing 'oxidative stress'. Nuclear poly-ADP-ribose (PAR) formation thought to be induced in response oxidative DNA damage promote cell death under sustained stress conditions. However, what exactly triggers PAR induction incompletely understood. Using reverse phase protein array (RPPA) in-depth analysis key signaling...
ABSTRACT The human genome encodes 45 kinesins that drive cell division, motility, intracellular trafficking, and ciliary function. Determining the cellular function of each kinesin would be greatly facilitated by specific small molecule inhibitors, but screens have yielded inhibitors are to only a number kinesins, likely due high conservation motor domain across superfamily. Here we present chemical-genetic approach engineer motors retain microtubule-dependent motility in absence inhibitor...
Current methods employed to evaluate PARP inhibitor \(PARPi) toxicity can be time consuming, have limited sensitivity, may not well suited for screening purposes, or yield results from cell population averages rather than single information.To overcome these limitations we provide a detailed protocol assess PARPi at multiple phenotypic levels based on high-content microscopy work ow.This approach takes two days and allows sensitive cycle resolved analyses of PARPi-evoked DNA damage response...