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18 results on '"Huber, Stefan T."'

1. Snapshots ofPseudomonas aeruginosaSOS response activation complex reveal structural prerequisites for LexA engagement and cleavage

Author

Vascon, Filippo, primary, Felice, Sofia De, additional, Gasparotto, Matteo, additional, Huber, Stefan T., additional, Catalano, Claudio, additional, Chinellato, Monica, additional, Grinzato, Alessandro, additional, Filippini, Francesco, additional, Maso, Lorenzo, additional, Jakobi, Arjen J., additional, and Cendron, Laura, additional

Published
2024
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6. Structural basis of antimicrobial membrane coat assembly by human GBP1

Author

Kuhm, Tanja, Taisne, Clémence, de Agrela Pinto, Cecilia, Gross, Luca, Giannopoulou, Evdokia A., Huber, Stefan T., Pardon, Els, Steyaert, Jan, Tans, Sander J., and Jakobi, Arjen J.

Abstract

Guanylate-binding proteins (GBPs) are interferon-inducible guanosine triphosphate hydrolases (GTPases) mediating host defense against intracellular pathogens. Their antimicrobial activity hinges on their ability to self-associate and coat pathogen-associated compartments or cytosolic bacteria. Coat formation depends on GTPase activity but how nucleotide binding and hydrolysis prime coat formation remains unclear. Here, we report the cryo-electron microscopy structure of the full-length human GBP1 dimer in its guanine nucleotide-bound state and describe the molecular ultrastructure of the GBP1 coat on liposomes and bacterial lipopolysaccharide membranes. Conformational changes of the middle and GTPase effector domains expose the isoprenylated C terminus for membrane association. The α-helical middle domains form a parallel, crossover arrangement essential for coat formation and position the extended effector domain for intercalation into the lipopolysaccharide layer of gram-negative membranes. Nucleotide binding and hydrolysis create oligomeric scaffolds with contractile abilities that promote membrane extrusion and fragmentation. Our data offer a structural and mechanistic framework for understanding GBP1 effector functions in intracellular immunity.

Published
2024
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14. Large-Scale Mapping of Axonal Arbors Using High-Density Microelectrode Arrays

Author

Bullmann, Torsten, Radivojevic, Milos, Huber, Stefan T., Deligkaris, Kosmas, Hierlemann, Andreas, and Frey, Urs

Subjects
action potential, action potential propagation, high-density microelectrode array, nervous system, extracellular electrical field, fungi, axons, axonal arborizations, high-throughput screening
Abstract

Understanding the role of axons in neuronal information processing is a fundamental task in neuroscience. Over the last years, sophisticated patch-clamp investigations have provided unexpected and exciting data on axonal phenomena and functioning, but there is still a need for methods to investigate full axonal arbors at sufficient throughput. Here, we present a new method for the simultaneous mapping of the axonal arbors of a large number of individual neurons, which relies on their extracellular signals that have been recorded with high-density microelectrode arrays (HD-MEAs). The segmentation of axons was performed based on the local correlation of extracellular signals. Comparison of the results with both, ground truth and receiver operator characteristics, shows that the new segmentation method outperforms previously used methods. Using a standard HD-MEA, we mapped the axonal arbors of 68 neurons in, Frontiers in Cellular Neuroscience, 13, ISSN:1662-5102

Published
2019

15. Structural basis of p62/SQSTM1 helical filaments, their presence in p62 bodies and role in cargo recognition in the cell

Author

Jakobi, Arjen J., primary, Huber, Stefan T., additional, Mortensen, Simon A., additional, Schultz, Sebastian W., additional, Palara, Anthimi, additional, Kuhm, Tanja, additional, Shrestha, Birendra Kumar, additional, Lamark, Trond, additional, Hagen, Wim J.H., additional, Wilmanns, Matthias, additional, Johansen, Terje, additional, Brech, Andreas, additional, and Sachse, Carsten, additional

Published
2019
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17. Structural basis of p62/SQSTM1 helical filaments and their role in cellular cargo uptake

Author

Jakobi, Arjen J., Huber, Stefan T., Mortensen, Simon A., Schultz, Sebastian W., Palara, Anthimi, Kuhm, Tanja, Shrestha, Birendra Kumar, Lamark, Trond, Hagen, Wim J. H., Wilmanns, Matthias, Johansen, Terje, Brech, Andreas, and Sachse, Carsten

Subjects
3. Good health
Abstract

Nature Communications 11(1), 440 (2020). doi:10.1038/s41467-020-14343-8, p62/SQSTM1 is an autophagy receptor and signaling adaptor with an N-terminal PB1 domain that forms the scaffold of phase-separated p62 bodies in the cell. The molecular determinants that govern PB1 domain filament formation in vitro remain to be determined and the role of p62 filaments inside the cell is currently unclear. We here determine four high-resolution cryo-EM structures of different human and Arabidopsis PB1 domain assemblies and observed a filamentous ultrastructure of p62/SQSTM1 bodies using correlative cellular EM. We show that oligomerization or polymerization, driven by a double arginine finger in the PB1 domain, is a general requirement for lysosomal targeting of p62. Furthermore, the filamentous assembly state of p62 is required for autophagosomal processing of the p62-specific cargo KEAP1. Our results show that using such mechanisms, p62 filaments can be critical for cargo uptake in autophagy and are an integral part of phase-separated p62 bodies., Published by Nature Publishing Group UK, [London]

18. Large-Scale Mapping of Axonal Arbors Using High-Density Microelectrode Arrays.

Author

Bullmann T, Radivojevic M, Huber ST, Deligkaris K, Hierlemann A, and Frey U

Abstract

Understanding the role of axons in neuronal information processing is a fundamental task in neuroscience. Over the last years, sophisticated patch-clamp investigations have provided unexpected and exciting data on axonal phenomena and functioning, but there is still a need for methods to investigate full axonal arbors at sufficient throughput. Here, we present a new method for the simultaneous mapping of the axonal arbors of a large number of individual neurons, which relies on their extracellular signals that have been recorded with high-density microelectrode arrays (HD-MEAs). The segmentation of axons was performed based on the local correlation of extracellular signals. Comparison of the results with both, ground truth and receiver operator characteristics, shows that the new segmentation method outperforms previously used methods. Using a standard HD-MEA, we mapped the axonal arbors of 68 neurons in <6 h. The fully automated method can be extended to new generations of HD-MEAs with larger data output and is estimated to provide data of axonal arbors of thousands of neurons within recording sessions of a few hours.

Published
2019
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