A5082422826- Nuclear Structure and Function
- RNA Research and Splicing
- Genomics and Chromatin Dynamics
- Microtubule and mitosis dynamics
- Force Microscopy Techniques and Applications
- Advanced Fluorescence Microscopy Techniques
- Advanced Electron Microscopy Techniques and Applications
- Reproductive Biology and Fertility
- Biotin and Related Studies
- Cellular transport and secretion
- DNA Repair Mechanisms
- Chromosomal and Genetic Variations
- Monoclonal and Polyclonal Antibodies Research
- Glycosylation and Glycoproteins Research
- Endoplasmic Reticulum Stress and Disease
- Bacteriophages and microbial interactions
- ATP Synthase and ATPases Research
- Molecular Communication and Nanonetworks
- Porphyrin Metabolism and Disorders
- RNA and protein synthesis mechanisms
- Evolution and Genetic Dynamics
- Trace Elements in Health
- RNA regulation and disease
- Telomeres, Telomerase, and Senescence
- Mitochondrial Function and Pathology
European Molecular Biology Laboratory
2017-2025
European Molecular Biology Laboratory
2014-2024
Medical University of Vienna
2019-2024
Vienna Biocenter
2021-2024
Max Perutz Labs
2019-2024
Kyoto University
2006-2013
The nuclear pore complex (NPC) mediates nucleocytoplasmic transport through the envelope. How NPC assembles into this double membrane boundary has remained enigmatic. Here, we captured temporally staged assembly intermediates by correlating live cell imaging with high-resolution electron tomography and super-resolution microscopy. Intermediates were dome-shaped evaginations of inner (INM), that grew in diameter depth until they fused flat outer membrane. Live super-resolved fluorescence...
Abstract Dividing eukaryotic cells package extremely long chromosomal DNA molecules into discrete bodies to enable microtubule-mediated transport of one genome copy each the newly forming daughter 1–3 . Assembly mitotic chromosomes involves looping by condensin 4–8 and chromatin compaction global histone deacetylation 9–13 Although confers mechanical resistance spindle pulling forces 14–16 , it is not known how affects material properties and, as a consequence, segregation mechanics...
Understanding how the nuclear pore complex (NPC) is assembled of fundamental importance to grasp mechanisms behind its essential function and understand role during evolution eukaryotes
Dynamic processes involving biomolecules are essential for the function of cell. Here, we introduce an integrative method computing models these based on multiple heterogeneous sources information, including time-resolved experimental data and physical dynamic processes. First, each time point, a set coarse compositional structural heterogeneity is computed (heterogeneity models). Second, model, static structure (a snapshot model). Finally, selected connected into series trajectories that...
Importin-beta mediates protein transport across the nuclear envelope through pore complex (NPC) by interacting with components of NPC, called nucleoporins, and a small G protein, Ran. Although there is accumulated knowledge on specific interaction between importin-beta Phe-Gly (FG) motif in nucleoporins as well effect RanGTP this interaction, molecular mechanism which shuttles NPC unknown. In study, we focused four binding pockets for FG motifs characterized using single-molecule...
Abstract Junctions between the endoplasmic reticulum (ER) and outer membrane of nuclear envelope (NE) physically connect both organelles. These ER–NE junctions are essential for supplying NE with lipids proteins synthesized in ER. However, little is known about structure these junctions. Here, we systematically study ultrastructure cryo-fixed mammalian cells staged anaphase, telophase, interphase by correlating live cell imaging three-dimensional electron microscopy. Our results show that...
The nuclear pore complex (NPC) is the only gateway for molecular trafficking across envelope. NPC not merely a static nuclear-cytoplasmic transport gate; functional analysis of nucleoporins has revealed dynamic features in various cellular functions, such as mitotic spindle formation and protein modification. However, it known whether undergoes changes during biological processes cell differentiation. In present study, we evaluate expression levels several show that amount Nup358/RanBP2...
Disulfide (S-S) bonds play important roles in the regulation of protein function and cellular stress responses. In this study, we demonstrate that distinct sets nucleoporins (Nups), components nuclear pore complex (NPC), form S-S regulate transport through NPC. Kinetic analysis importin β demonstrated permeability NPC was increased by dithiothreitol treatment reduced oxidative stress. The small proteins such as GFP not affected either or a reducing reagent. Immunoblot revealed significantly...
The accuracy and the fidelity of a single‐molecule force measurement largely rely on how molecule interest is attached to solid substrate surface (bead, cantilever, cover glass etc.). A site‐specific attachment protein without affecting its structure enzymatic function has been major concern. Here, we established glutathione‐coupled cantilever which any glutathione S ‐transferase (GST)‐fused proteins can be in desired direction. rupture between GST was ∼100 pN average. By using this...
Different types of anaphase bridges are reported to form between segregating chromosomes during cell division. Previous studies using laser microsurgery suggested that elastic tethers connect the telomeres separating in many animal meiotic and mitotic cells. However, structural evidence is lacking for their existence. In this study, by correlating live imaging with electron tomography, we examined whether visible structures meiosis I crane-fly primary spermatocytes. We found extending all...
Karyopherin-dependent molecular transport through the nuclear pore complex is maintained by constant recycling pathways of karyopherins coupled with Ran-dependent cargo catch-and-release mechanism. Although many studies have revealed bidirectional dynamics karyopherins, entire kinetics steady-state karyopherin and still not fully understood. In this study, we used fluorescence recovery after photobleaching loss in on live cells to provide convincing vivo proof that karyopherin-mediated...
ABSTRACT We introduce an engineered nanobody whose affinity to green fluorescent protein (GFP) can be switched on and off with small molecules. By controlling the cellular localization of GFP fusion proteins, allows study their role in basic biological processes, approach that should applicable numerous previously described fusions. also outline how binding affinities other nanobodies controlled by
Abstract Dividing eukaryotic cells package extremely long chromosomal DNA molecules into discrete bodies to enable microtubule-mediated transport of one genome copy each the newly forming daughter 1–3 . Assembly mitotic chromosomes involves looping by condensin 4–8 and chromatin compaction global histone deacetylation 9–13 While confers mechanical resistance towards spindle pulling forces 14–16 , it is not known how affects material properties segregation mechanics chromosomes. Here, we show...
Abstract The endoplasmic reticulum (ER) is physically connected to the nucleus by junctions with outer membrane of nuclear envelope (NE). ER–NE are essential for supplying NE lipids and proteins synthesized in ER. However, little known about structure these junctions. Here, we systematically studied ultrastructure cryo-fixed mammalian cells staged anaphase, telophase, interphase correlating live cell imaging three-dimensional electron microscopy. Strikingly, our results revealed that have a...
Summary Understanding how the nuclear pore complex (NPC) assembles is of fundamental importance to grasp mechanisms behind its essential function and understand role during evolution eukaryotes 1–4 . While at least two NPC assembly pathways exist, one exit from mitosis growth in interphase, we currently lack a quantitative map molecular events. Here, use fluorescence correlation spectroscopy (FCS) calibrated live imaging endogenously fluorescently-tagged nucleoporins changes composition...
Cellular communication: unlocking the secrets of ER-NE junctions The conNEctoER project aims to reveal mechanism governing essential within our cells, that supply proteins and lipids cell nucleus. By combining electron microscopy live imaging, we plan elucidate junction formation function in human shedding light on gene expression, nuclear organisation disease mechanisms.