A5026411335- Epigenetics and DNA Methylation
- CRISPR and Genetic Engineering
- Genomics and Chromatin Dynamics
- RNA modifications and cancer
- Cancer-related gene regulation
- Advanced biosensing and bioanalysis techniques
- Pluripotent Stem Cells Research
- Histone Deacetylase Inhibitors Research
- RNA and protein synthesis mechanisms
- Acute Myeloid Leukemia Research
- RNA Research and Splicing
- RNA Interference and Gene Delivery
- Ubiquitin and proteasome pathways
- Inflammation biomarkers and pathways
- Protein Degradation and Inhibitors
- Monoclonal and Polyclonal Antibodies Research
- Genetics and Neurodevelopmental Disorders
- Chromatin Remodeling and Cancer
- Neuroinflammation and Neurodegeneration Mechanisms
- Neurological Disease Mechanisms and Treatments
- Genetic Syndromes and Imprinting
- Alzheimer's disease research and treatments
- Immune Cell Function and Interaction
- HIV Research and Treatment
- Chromosomal and Genetic Variations
Ludwig-Maximilians-Universität München
2015-2025
Center for Integrated Protein Science Munich
2012-2021
Faculty (United Kingdom)
2021
Urologische Klinik München
2014
The recent discovery of genomic 5-hydroxymethylcytosine (hmC) and mutations affecting the respective Tet hydroxylases in leukemia raises fundamental questions about this epigenetic modification. We present a sensitive method for fast quantification hmC based on specific transfer radiolabeled glucose to by purified glucosyltransferase. determined levels various adult tissues differentiating embryonic stem cells show correlation with differential expression tet genes.
Epigenome-wide association studies (EWAS) based on human brain samples allow a deep and direct understanding of epigenetic dysregulation in Alzheimer's disease (AD). However, strong variation cell-type proportions across tissue represents significant source data noise. Here, we report the first EWAS sorted neuronal non-neuronal (mostly glia) nuclei from postmortem tissues.We show that cell sorting strongly enhances robust detection disease-related DNA methylation changes even relatively...
Specific control of gene activity is a valuable tool to study and engineer cellular functions. Recent studies uncovered the potential transcription activator-like effector (TALE) proteins that can be tailored activate user-defined target genes. It remains however unclear whether how epigenetic modifications interfere with TALE-mediated transcriptional activation. We studied five designer TALEs (dTALEs) targeting oct4 pluripotency gene. In vitro assays showed dTALEs distinct sites in promoter...
Recent studies have indicated that nuclear protein of 95 kDa (Np95) is essential for maintaining genomic methylation by recruiting DNA methyltransferase (Dnmt) 1 to hemi-methylated sites. Here, we show Np95 interacts more strongly with regulatory domains the de novo methyltransferases Dnmt3a and Dnmt3b. To investigate possible functions, developed an epigenetic silencing assay using fluorescent reporters in embryonic stem cells (ESCs). Interestingly, cytomegalovirus promoter ESCs preceded...
Recent discovery of 5-hydroxymethylcytosine (5hmC) in genomic DNA raises the question how this sixth base is recognized by cellular proteins. In contrast to methyl-CpG binding domain (MBD) MeCP2, we found that SRA Uhrf1, an essential factor maintenance methylation, binds 5hmC and 5-methylcytosine containing substrates with similar affinity. Based on co-crystal structure, performed molecular dynamics simulations SRA:DNA complex flipped cytosine carrying either these epigenetic modifications....
Labeling and tracing of specific sequences in living cells has been a major challenge studying the spatiotemporal dynamics native chromatin. Here we repurposed prokaryotic CRISPR/Cas adaptive immunity system to specifically detect endogenous genomic loci mouse embryonic stem cells. We constructed catalytically inactive version Cas9 endonuclease, fused it with eGFP (dCas9-eGFP) co-expressed small guide RNAs (gRNAs) target pericentric, centric, telomeric repeats, which are enriched distinct...
Covalent chemical modifications of cellular RNAs directly impact all biological processes. However, our mechanistic understanding the enzymes catalyzing these modifications, their substrates and functions, remains vague. Amongst RNA N 6 -methyladenosine (m A) is widespread found in messenger (mRNA), ribosomal (rRNA), noncoding RNAs. Here, we undertook a systematic screen to uncover new methyltransferases. We demonstrate that methyltransferase-like 5 (METTL5) protein catalyzes m A 18S rRNA at...
Chromatin relaxation is one of the earliest cellular responses to DNA damage. However, what determines these structural changes, including their ATP requirement, not well understood. Using live-cell imaging and laser microirradiation induce lesions, we show that local chromatin at damage sites regulated by PARP1 enzymatic activity. We also report H1 mobilized sites, but, since this mobilization largely independent poly(ADP-ribosyl)ation, it cannot solely explain relaxation. Finally,...
Stable inheritance of DNA methylation is critical for maintaining differentiated phenotypes in multicellular organisms. We have recently identified dual mono-ubiquitylation histone H3 (H3Ub2) by UHRF1 as an essential mechanism to recruit DNMT1 chromatin. Here, we show that PCNA-associated factor 15 (PAF15) undergoes UHRF1-dependent (PAF15Ub2) on chromatin a replication-coupled manner. This event will, turn, DNMT1. During early S-phase, preferentially ubiquitylates PAF15, whereas H3Ub2...
Research Article23 May 2019Open Access Source DataTransparent process Opposite microglial activation stages upon loss of PGRN or TREM2 result in reduced cerebral glucose metabolism Julia K Götzl Chair Metabolic Biochemistry, Biomedical Center (BMC), Faculty Medicine, Ludwig-Maximilians-Universität München, Munich, Germany Search for more papers by this author Matthias Brendel Department Nuclear University Hospital, Georg Werner Samira Parhizkar Laura Sebastian Monasor German...
Laron syndrome (LS) is a rare, autosomal recessive disorder in humans caused by loss-of-function mutations of the growth hormone receptor (GHR) gene. To establish large animal model for LS, pigs with GHR knockout (KO) were generated and characterized.
RNA methyltransferase METTL6 is implicated in tumor cell growth and mouse energy consumption.
Several mammalian proteins involved in chromatin and DNA modification contain CXXC zinc finger domains. We compared the structure function of domains methyltransferase Dnmt1 methylcytosine dioxygenase Tet1. Sequence alignment showed that both have a very similar framework but differ central tip region. Based on known MLL1 domain we developed homology models designed expression constructs for isolated Tet1 accordingly. show has no binding activity is dispensable catalytic vivo. In contrast,...
The RING E3 ubiquitin ligase UHRF1 controls DNA methylation through its ability to target the maintenance methyltransferase DNMT1 newly replicated chromatin. recruitment relies on ubiquitylation of histone H3 by UHRF1; however, how deposits onto is unknown. Here, we demonstrate that ubiquitin-like domain (UBL) essential for RING-mediated ubiquitylation. Using chemical crosslinking and mass spectrometry, biochemical assays, recombinant chromatin substrates, show UBL participates in structural...
Acute myeloid leukemia (AML) is an aggressive hematologic neoplasm resulting from the malignant transformation of progenitors. Despite intensive chemotherapy leading to initial treatment responses, relapse caused by intrinsic or acquired drug resistance represents a major challenge. Here, we report that histone 3 lysine 27 demethylase KDM6A (UTX) targeted inactivating mutations and mutation-independent regulation in relapsed AML. Analyses matched diagnosis specimens individuals with showed...
Tumor cells orchestrate their microenvironment. Here, we provide biochemical, structural, functional, and clinical evidence that Cathepsin S (CTSS) alterations induce a tumor-promoting immune microenvironment in follicular lymphoma (FL). We found CTSS mutations at Y132 6% of FL (19/305). Another 13% (37/286) had amplification, which was associated with higher expression. lead to accelerated autocatalytic conversion from an enzymatically inactive profrom active increased substrate cleavage,...
Abstract Genome-wide DNA demethylation is a unique feature of mammalian development and naïve pluripotent stem cells. Here, we describe recently evolved pathway in which global hypomethylation achieved by the coupling active passive demethylation. TET activity required, albeit indirectly, for demethylation, mostly occurs at sites devoid binding. Instead, TET-mediated locus-specific necessary activating subset genes, including pluripotency germline marker Dppa3 ( Stella, Pgc7 ). DPPA3 turn...
The genetic code of mammalian cells can be expanded to allow the incorporation non-canonical amino acids (ncAAs) by suppressing in-frame amber stop codons (UAG) with an orthogonal pyrrolysyl-tRNA synthetase (PylRS)/tRNAPylCUA (PylT) pair. However, feasibility this approach is substantially hampered unpredictable variations in efficiencies at different codon positions within target proteins. Here, we apply a proteomics-based quantify ncAA rates hundreds endogenous cells. With these data,...
Abstract Epigenetic regulation of gene expression involves, besides DNA and histone modifications, the relative positioning sequences within nucleus. To trace specific in living cells, we used programmable sequence-specific binding designer transcription activator-like effectors (dTALEs). We designed a recombinant dTALE (msTALE) with variable repeat domains to specifically bind 19-bp target sequence major satellite DNA. The msTALE was fused green fluorescent protein (GFP) stably expressed...
Chemotherapy resistance is the main impediment in treatment of acute myeloid leukaemia (AML). Despite rapid advances, various mechanisms inducing development remain to be defined detail. Here we report that loss-of-function mutations (LOF) histone methyltransferase EZH2 have potential confer against chemotherapeutic agent cytarabine. We identify seven distinct leading loss H3K27 trimethylation via multiple mechanisms. Analysis matched diagnosis and relapse samples reveal a heterogenous...