A5056598193- DNA Repair Mechanisms
- Microtubule and mitosis dynamics
- Genomics and Chromatin Dynamics
- RNA and protein synthesis mechanisms
- Chromosomal and Genetic Variations
- RNA modifications and cancer
- Bacterial Genetics and Biotechnology
- CRISPR and Genetic Engineering
- Fungal and yeast genetics research
- Mitochondrial Function and Pathology
- RNA Research and Splicing
- Photosynthetic Processes and Mechanisms
- Viral Infectious Diseases and Gene Expression in Insects
- Epigenetics and DNA Methylation
- Immune Response and Inflammation
- Physiological and biochemical adaptations
- Retinal Development and Disorders
- Ubiquitin and proteasome pathways
- Protein purification and stability
- Endoplasmic Reticulum Stress and Disease
- Monoclonal and Polyclonal Antibodies Research
- Neonatal and Maternal Infections
- Neonatal Respiratory Health Research
- Cancer Mechanisms and Therapy
- Zebrafish Biomedical Research Applications
Friedrich Miescher Laboratory
2017-2024
Max Planck Society
2020-2024
Max Planck Institute of Molecular Physiology
2013-2020
Max Planck Institute of Biochemistry
2010-2014
MRC Laboratory of Molecular Biology
2006-2009
The crystal structure of the bacterial 70S ribosome refined to 2.8 angstrom resolution reveals atomic details its interactions with messenger RNA (mRNA) and transfer (tRNA). A metal ion stabilizes a kink in mRNA that demarcates boundary between P sites, which is potentially important prevent slippage mRNA. Metal ions also stabilize intersubunit interface. E-site tRNA 50S subunit have both similarities differences compared those archaeal ribosome. rationalizes much biochemical genetic data on...
The ribosome selects a correct transfer RNA (tRNA) for each amino acid added to the polypeptide chain, as directed by messenger RNA. Aminoacyl-tRNA is delivered elongation factor Tu (EF-Tu), which hydrolyzes guanosine triphosphate (GTP) and releases tRNA in response codon recognition. signaling pathway that leads GTP hydrolysis upon recognition critical accurate decoding. Here we present crystal structure of complexed with EF-Tu aminoacyl-tRNA, refined 3.6 angstrom resolution. reveals...
Regulation of macromolecular interactions by phosphorylation is crucial in signaling networks. In the spindle assembly checkpoint (SAC), which enables errorless chromosome segregation, promotes recruitment SAC proteins to tensionless kinetochores. The kinase Mps1 phosphorylates multiple Met-Glu-Leu-Thr (MELT) motifs on kinetochore subunit Spc105/Knl1. phosphorylated MELT (MELTP) then promote downstream components. How MELTP are recognized unclear. this study, we report that Bub3, a 7-bladed...
Flagellin, the protein subunit of bacterial flagellum, stimulates innate immune receptor Toll-like 5 (TLR5) after pattern recognition or evades TLR5 through lack recognition. This binary response fails to explain weak agonism flagellins from commensal bacteria, raising question how is tuned. Here, we screened abundant present in metagenomes human gut for both and activation uncovered a class flagellin-TLR5 interaction termed silent Silent were agonists despite Receptor activity was tuned by...
Kinetochores are multisubunit complexes that assemble on centromeres to bind spindle microtubules and promote faithful chromosome segregation during cell division. A 16-subunit complex named the constitutive centromere–associated network (CCAN) creates centromere–kinetochore interface. CENP-C, a CCAN subunit, is crucial for kinetochore assembly because it links with microtubule-binding interface of kinetochores. The role CENP-C in organization, other hand, had been incompletely understood....
Mtr4 is a conserved RNA helicase that functions together with the nuclear exosome. It participates in processing of structured RNAs, including maturation 5.8S ribosomal (rRNA). also interacts polyadenylating Trf4-Air2 heterodimer to form so-called TRAMP ( Tr f4- A ir2- M tr4 P olyadenylation) complex. involved exosome-mediated degradation aberrant RNAs surveillance pathways. We report 2.9-Å resolution crystal structure Saccharomyces cerevisiae complex ADP and RNA. The shows central ATPase...
Kinetochores, multi-subunit complexes that assemble at the interface with centromeres, bind spindle microtubules to ensure faithful delivery of chromosomes during cell division. The configuration and function kinetochore-centromere is poorly understood. We report a protein this interface, CENP-M, structurally evolutionarily related small GTPases but incapable GTP-binding conformational switching. show CENP-M crucially required for assembly stability tetramer also comprising CENP-I, CENP-H,...
Centromere protein (CENP) A, a histone H3 variant, is key epigenetic determinant of chromosome domains known as centromeres. Centromeres nucleate kinetochores, multi-subunit complexes that capture spindle microtubules to promote segregation during mitosis. Two kinetochore proteins, CENP-C and CENP-N, recognize CENP-A in the context rare nucleosome. Here, we reveal structural basis for exquisite selectivity CENP-N uses charge space complementarity decode L1 loop unique CENP-A. It also engages...
Abstract Programmed DNA double-strand break (DSB) formation is a crucial feature of meiosis in most organisms. DSBs initiate recombination-mediated linking homologous chromosomes, which enables correct chromosome segregation meiosis. are generated on axes by heterooligomeric focal clusters DSB-factors. Whereas DNA-driven protein condensation thought to assemble the DSB-machinery, its targeting poorly understood. We uncover mice that efficient biogenesis DSB-machinery requires seeding axial...
The approximately thirty core subunits of kinetochores assemble on centromeric chromatin containing the histone H3 variant CENP-A and connect chromosomes with spindle microtubules. proximal 16-subunit CCAN (constitutive centromere associated network) creates a mechanically stable bridge between kinetochore's microtubule-binding machinery, 10-subunit KMN assembly. Here, we reconstituted stoichiometric 11-subunit human that forms when CENP-OPQUR complex binds to joint interface CENP-HIKM...
In meiosis, DNA double-strand break (DSB) formation by Spo11 initiates recombination and enables chromosome segregation. Numerous factors are required for activity, couple the DSB machinery to development of a meiosis-specific 'axis-tethered loop' organisation. Through in vitro reconstitution budding yeast genetics, we here provide architectural insight into focussing on foundational factor, Mer2. We characterise interaction Mer2 with histone reader Spp1, show that directly associates...
The spindle assembly checkpoint (SAC) monitors microtubule attachment to kinetochores ensure accurate sister chromatid segregation during mitosis. SAC members Bub1 and BubR1 are paralogs that underwent significant functional specializations evolution. We report an in-depth characterization of the kinase domains BubR1. domain binds nucleotides but is unable deliver catalytic activity in vitro. Conversely, active regulated by intra-molecular phosphorylation at P+1 loop. crystal structure...
Abstract Crossing over between homologs is critical for the stable segregation of chromosomes during first meiotic division. Saccharomyces cerevisiae Mer3 (HFM1 in mammals) a SF2 helicase and member ZMM group proteins, that facilitates formation majority crossovers meiosis. Here, we describe structural organisation using AlphaFold modelling XL-MS further characterise previously described interaction with Mlh1–Mlh2. We find also forms undescribed complex recombination regulating factors Top3...
Abstract The successful production of recombinant protein for biochemical, biophysical, and structural biological studies critically depends on the correct expression organism. Currently, most commonly used organisms are Escherichia coli (~70% all PDB structures) baculovirus/ insect cell system (~5% structures). While is frequently large eukaryotic proteins, it relatively expensive time‐consuming compared to E. expression. Frequently decision carry out a baculovirus project means restarting...
For meiosis I, homologous chromosomes must be paired into bivalents. Maintenance of homolog conjunction in bivalents until anaphase I depends on crossovers canonical meiosis. However, instead crossovers, an alternative system achieves during the achiasmate male Drosophila melanogaster . The proteins SNM, UNO and MNM are likely constituents a physical linkage that conjoins homologs D spermatocytes. Here, we report SNM binds tightly to C-terminal region UNO. This interaction is cohesin...
Summary Programmed DNA double-strand break (DSB) formation is a unique meiotic feature that initiates recombination-mediated linking of homologous chromosomes, thereby enabling chromosome number halving in meiosis. DSBs are generated on axes by heterooligomeric focal clusters DSB-factors. Whereas DNA-driven protein condensation thought to assemble the DSB-machinery, its targeting poorly understood. We discovered mice efficient biogenesis DSB-machinery requires seeding axial IHO1 platforms,...
The bacterial protein flagellin is the sole ligand of innate immune receptor Toll-like 5 (TLR5). Flagellins with strong agonism bind TLR5 at their D1 and D0 domains, while poor agonist silent flagellins only. However, D0-TLR5 interactions insufficiently explain phenotype. Here, we characterize domain binding kinetics RhFlaB compared to canonical StFliC. Using Surface Plasmon Resonance, show that RhFlaB-D1 binds more strongly than StFliC-D1, but forms shorter-lived complexes. Cryo-EM analysis...
Teleost fish of the genus Danio are excellent models to study genetic and cellular bases pigment pattern variation in vertebrates. The two sister species rerio aesculapii show divergent patterns horizontal stripes vertical bars that partly caused by divergence potassium channel gene kcnj13. Here, we kcnj13 is required only melanophores for interactions with xanthophores iridophores, which cause location-specific cell shapes thereby influence colour contrast D. rerio. Cis-regulatory rather...
Pch2 is a meiosis-specific AAA+ protein that controls several important chromosomal processes. We previously demonstrated Orc1, subunit of the ORC, functionally interacts with budding yeast Pch2. The ORC (Orc1-6) complex loads MCM helicase to origins replication, but whether and how collaborates remains unclear. Here, we show hexamer directly associates during meiotic G2/prophase. Biochemical analysis suggests uses its non-enzymatic NH 2 -terminal domain core likely engages interface also...
In meiosis, DNA double strand break (DSB) formation by Spo11 initiates recombination and enables chromosome segregation. Numerous factors are required for activity, couple the DSB machinery to development of a meiosis-specific ?axis-tethered loop? organization. Through in vitro reconstitution budding yeast genetics we here provide architectural insight into focussing on foundational factor, Mer2. We characterise interaction Mer2 with histone reader Spp1, show that directly associates...