A5078170930- Nuclear Structure and Function
- RNA Research and Splicing
- Genomics and Chromatin Dynamics
- Cellular transport and secretion
- RNA and protein synthesis mechanisms
- Advanced Electron Microscopy Techniques and Applications
- RNA modifications and cancer
- Advanced Fluorescence Microscopy Techniques
- Transgenic Plants and Applications
- RNA Interference and Gene Delivery
- Photosynthetic Processes and Mechanisms
- Monoclonal and Polyclonal Antibodies Research
- Bacteriophages and microbial interactions
- Cancer Genomics and Diagnostics
- Near-Field Optical Microscopy
- Forensic and Genetic Research
- SARS-CoV-2 and COVID-19 Research
- Silk-based biomaterials and applications
- Biochemical and Structural Characterization
- Microtubule and mitosis dynamics
- Electron Spin Resonance Studies
- CRISPR and Genetic Engineering
Max Planck Institute for Multidisciplinary Sciences
2022-2024
Max Planck Institute for Biophysical Chemistry
2020-2022
Leibniz Association
2013
Monoclonal anti-SARS-CoV-2 immunoglobulins represent a treatment option for COVID-19. However, their production in mammalian cells is not scalable to meet the global demand. Single-domain (VHH) antibodies (also called nanobodies) provide an alternative suitable microbial production. Using alpaca immune libraries against receptor-binding domain (RBD) of SARS-CoV-2 Spike protein, we isolated 45 infection-blocking VHH antibodies. These include nanobodies that can withstand 95°C. The most...
Abstract Super-resolution techniques have achieved localization precisions in the nanometer regime. Here we report all-optical, room temperature of fluorophores with precision Ångström range. We built on concept MINSTED nanoscopy where is increased by encircling fluorophore low-intensity central region a stimulated emission depletion (STED) donut beam while constantly increasing absolute power. By blue-shifting STED and separating on/off switching, individual bound to DNA strand are...
In eukaryotes, RNA Polymerase (Pol) III is specialized for the transcription of tRNAs and other short, untranslated RNAs. Pol a determinant cellular growth lifespan across eukaryotes. Upregulation observed in cancer causative mutations have been described neurodevelopmental disorders hypersensitivity to viral infection. Here, we report cryo-EM reconstruction at 4.0 Å human III, allowing mapping rationalization reported genetic mutations. Mutations causing defects cluster hotspots affecting...
The proteins ZC3HC1 and TPR are structural components of the nuclear basket (NB), a fibrillar structure attached to nucleoplasmic side pore complex (NPC). initially binds NB in TPR-dependent manner can subsequently recruit additional polypeptides this structure. Here, we examined molecular properties that enable its initial binding TPR. We report identification definition basket-interaction domain (NuBaID)
Abstract Nuclear pore complex (NPC) biogenesis is a still enigmatic example of protein self-assembly. We now introduce several cross-reacting anti-Nup nanobodies for imaging intact nuclear complexes from frog to human. also report simplified assay that directly tracks postmitotic NPC assembly with added fluorophore-labeled nanobodies. During interphase, NPCs are inserted into pre-existing envelope. Monitoring this process challenging because newly assembled indistinguishable ones. overcame...
The nuclear basket (NB) scaffold, a fibrillar structure anchored to the pore complex (NPC), is regarded as constructed of polypeptides coiled-coil dominated protein TPR which other proteins can bind without contributing NB's structural integrity. Here we report vertebrate ZC3HC1 novel inherent constituent NB, common at envelopes (NE) proliferating and non-dividing, terminally differentiated cells different morphogenetic origin. Formerly described functions, instead present NB component...
The nuclear basket (NB), anchored to the pore complex (NPC), is commonly looked upon as a structure built solely of protein TPR polypeptides, latter thus regarded NB's only scaffold-forming components. In current study, we report ZC3HC1 second structural element NB. Recently described an NB-appended omnipresent in vertebrates, now show that ZC3HC1, both vivo and vitro, enables stepwise manner recruitment subpopulations NB their linkage already NPC-anchored polypeptides. We further...
In the last two decades it was shown that plants have a great potential for production of specific heterologous proteins. But high cost and inefficient downstream processing are main technical bottleneck broader use plant-based technology especially protein-based products, as fibres or biodegradable plastics also medical applications. High-performance from recombinant spider silks are, therefore, prominent example. Spiders developed rather different silk materials based on These show...
We report all-optical, room-temperature localization of fluorophores with precision in the Ångström range. These precisions are attained a STED microscope, by encircling fluorophore low-intensity edge donut beam, while constantly increasing absolute power. Individual bound to DNA strand localized σ = 4.7 Å, corresponding fraction size, only 2,000 detected photons. MINSTED fluorescence nanoscopy single-digit nanometer resolution is exemplified imaging nuclear pore complexes and distribution...
Abstract The nuclear basket (NB), anchored to the pore complex (NPC), is commonly thought of as built solely protein TPR polypeptides, latter thus regarded NB’s only scaffold-forming components. In current study, we report ZC3HC1 a second building element NB. Recently described an NB-appended omnipresent in vertebrates, now show that ZC3HC1, both vivo and vitro , enables step-wise fashion recruitment subpopulations NB their linkage already NPC-anchored polypeptides. We further demonstrate...
ABSTRACT In eukaryotes, RNA Polymerase (Pol) III is the enzyme specialised for transcription of entire pool tRNAs and several other short, essential, untranslated RNAs. Pol a critical determinant cellular growth lifespan across eukaryotic kingdom. Upregulation often observed in cancer cells causative mutations have been described patients affected by severe neurodevelopmental disorders hypersensitivity to viral infection. Harnessing CRISPR-Cas9 genome editing HeLa cells, we isolated...
Abstract Proteins ZC3HC1 and TPR are construction elements of the nuclear pore complex (NPC)-attached basket (NB). NB-location depends on already occurring NPC-anchored, whereas additional polypeptides appended to NB by ZC3HC1. The current study examined molecular properties that enable it bind TPR. We report identification definition a basket-interaction domain (NuBaID) Hs comprising two similarly built modules, both essential for binding NB’s NPC-anchored Furthermore, we describe such...