61 results on '"Henning U"'
Search Results
2. Apoε4 disrupts neurovascular regulation and undermines white matter integrity and cognitive function
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Kenzo Koizumi, Yorito Hattori, Sung Ji Ahn, Izaskun Buendia, Antonio Ciacciarelli, Ken Uekawa, Gang Wang, Abigail Hiller, Lingzhi Zhao, Henning U. Voss, Steven M. Paul, Chris Schaffer, Laibaik Park, and Costantino Iadecola
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Science - Abstract
ApoE4 is a risk factor for small vessel disease, which can lead to cognitive impairment. Here the authors assess the microvasculature of the corpus callosum using 3-photon microscopy and find that mice expressing the ApoE4 allele are more susceptible than wild-type to white matter injury and cognitive impairment in a model of hypoperfusion-induced hypoxia.
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- 2018
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3. Sexual dimorphism in striatal dopaminergic responses promotes monogamy in social songbirds
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Kirill Tokarev, Julia Hyland Bruno, Iva Ljubičić, Paresh J Kothari, Santosh A Helekar, Ofer Tchernichovski, and Henning U Voss
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zebra finch ,dopamine ,social behavior ,monogamy ,sexual dimorphism ,reward circuitry ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
In many songbird species, males sing to attract females and repel rivals. How can gregarious, non-territorial songbirds such as zebra finches, where females have access to numerous males, sustain monogamy? We found that the dopaminergic reward circuitry of zebra finches can simultaneously promote social cohesion and breeding boundaries. Surprisingly, in unmated males but not in females, striatal dopamine neurotransmission was elevated after hearing songs. Behaviorally too, unmated males but not females persistently exchanged mild punishments in return for songs. Song reinforcement diminished when dopamine receptors were blocked. In females, we observed song reinforcement exclusively to the mate’s song, although their striatal dopamine neurotransmission was only slightly elevated. These findings suggest that song-triggered dopaminergic activation serves a dual function in social songbirds: as low-threshold social reinforcement in males and as ultra-selective sexual reinforcement in females. Co-evolution of sexually dimorphic reinforcement systems can explain the coexistence of gregariousness and monogamy.
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- 2017
- Full Text
- View/download PDF
4. Searching for Conservation Laws in Brain Dynamics—BOLD Flux and Source Imaging
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Henning U. Voss and Nicholas D. Schiff
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brain dynamics ,brain networking ,conservation law ,functional MRI ,fMRI ,mental imagery ,motor imagery ,synchronization ,Science ,Astrophysics ,QB460-466 ,Physics ,QC1-999 - Abstract
Blood-oxygen-level-dependent (BOLD) imaging is the most important noninvasive tool to map human brain function. It relies on local blood-flow changes controlled by neurovascular coupling effects, usually in response to some cognitive or perceptual task. In this contribution we ask if the spatiotemporal dynamics of the BOLD signal can be modeled by a conservation law. In analogy to the description of physical laws, which often can be derived from some underlying conservation law, identification of conservation laws in the brain could lead to new models for the functional organization of the brain. Our model is independent of the nature of the conservation law, but we discuss possible hints and motivations for conservation laws. For example, globally limited blood supply and local competition between brain regions for blood might restrict the large scale BOLD signal in certain ways that could be observable. One proposed selective pressure for the evolution of such conservation laws is the closed volume of the skull limiting the expansion of brain tissue by increases in blood volume. These ideas are demonstrated on a mental motor imagery fMRI experiment, in which functional brain activation was mapped in a group of volunteers imagining themselves swimming. In order to search for local conservation laws during this complex cognitive process, we derived maps of quantities resulting from spatial interaction of the BOLD amplitudes. Specifically, we mapped fluxes and sources of the BOLD signal, terms that would appear in a description by a continuity equation. Whereas we cannot present final answers with the particular analysis of this particular experiment, some results seem to be non-trivial. For example, we found that during task the group BOLD flux covered more widespread regions than identified by conventional BOLD mapping and was always increasing during task. It is our hope that these results motivate more work towards the search for conservation laws in neuroimaging experiments or at least towards imaging procedures based on spatial interactions of signals. The payoff could be new models for the dynamics of the healthy brain or more sensitive clinical imaging approaches, respectively.
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- 2014
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5. Altered auditory BOLD response to conspecific birdsong in zebra finches with stuttered syllables.
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Henning U Voss, Delanthi Salgado-Commissariat, and Santosh A Helekar
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Medicine ,Science - Abstract
How well a songbird learns a song appears to depend on the formation of a robust auditory template of its tutor's song. Using functional magnetic resonance neuroimaging we examine auditory responses in two groups of zebra finches that differ in the type of song they sing after being tutored by birds producing stuttering-like syllable repetitions in their songs. We find that birds that learn to produce the stuttered syntax show attenuated blood oxygenation level-dependent (BOLD) responses to tutor's song, and more pronounced responses to conspecific song primarily in the auditory area field L of the avian forebrain, when compared to birds that produce normal song. These findings are consistent with the presence of a sensory song template critical for song learning in auditory areas of the zebra finch forebrain. In addition, they suggest a relationship between an altered response related to familiarity and/or saliency of song stimuli and the production of variant songs with stuttered syllables.
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- 2010
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6. Defective mitochondrial COX1 translation due to loss of COX14 function triggers ROS-induced inflammation in mouse liver
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Abhishek Aich, Angela Boshnakovska, Steffen Witte, Tanja Gall, Kerstin Unthan-Fechner, Roya Yousefi, Arpita Chowdhury, Drishan Dahal, Aditi Methi, Svenja Kaufmann, Ivan Silbern, Jan Prochazka, Zuzana Nichtova, Marcela Palkova, Miles Raishbrook, Gizela Koubkova, Radislav Sedlacek, Simon E. Tröder, Branko Zevnik, Dietmar Riedel, Susann Michanski, Wiebke Möbius, Philipp Ströbel, Christian Lüchtenborg, Patrick Giavalisco, Henning Urlaub, Andre Fischer, Britta Brügger, Stefan Jakobs, and Peter Rehling
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Science - Abstract
Abstract Mitochondrial oxidative phosphorylation (OXPHOS) fuels cellular ATP demands. OXPHOS defects lead to severe human disorders with unexplained tissue specific pathologies. Mitochondrial gene expression is essential for OXPHOS biogenesis since core subunits of the complexes are mitochondrial-encoded. COX14 is required for translation of COX1, the central mitochondrial-encoded subunit of complex IV. Here we describe a COX14 mutant mouse corresponding to a patient with complex IV deficiency. COX14M19I mice display broad tissue-specific pathologies. A hallmark phenotype is severe liver inflammation linked to release of mitochondrial RNA into the cytosol sensed by RIG-1 pathway. We find that mitochondrial RNA release is triggered by increased reactive oxygen species production in the deficiency of complex IV. Additionally, we describe a COA3Y72C mouse, affected in an assembly factor that cooperates with COX14 in early COX1 biogenesis, which displays a similar yet milder inflammatory phenotype. Our study provides insight into a link between defective mitochondrial gene expression and tissue-specific inflammation.
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- 2024
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7. Correction to 'The thymocyte-specific RNA-binding protein Arpp21 provides TCR repertoire diversity by binding to the 3’-UTR and promoting Rag1 mRNA expression'
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Meng Xu, Taku Ito-Kureha, Hyun-Seo Kang, Aleksandar Chernev, Timsse Raj, Kai P. Hoefig, Christine Hohn, Florian Giesert, Yinhu Wang, Wenliang Pan, Natalia Ziętara, Tobias Straub, Regina Feederle, Carolin Daniel, Barbara Adler, Julian König, Stefan Feske, George C. Tsokos, Wolfgang Wurst, Henning Urlaub, Michael Sattler, Jan Kisielow, F. Gregory Wulczyn, Marcin Łyszkiewicz, and Vigo Heissmeyer
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Science - Published
- 2024
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8. PRPF8-mediated dysregulation of hBrr2 helicase disrupts human spliceosome kinetics and 5´-splice-site selection causing tissue-specific defects
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Robert Atkinson, Maria Georgiou, Chunbo Yang, Katarzyna Szymanska, Albert Lahat, Elton J. R. Vasconcelos, Yanlong Ji, Marina Moya Molina, Joseph Collin, Rachel Queen, Birthe Dorgau, Avril Watson, Marzena Kurzawa-Akanbi, Ross Laws, Abhijit Saxena, Chia Shyan Beh, Chileleko Siachisumo, Franziska Goertler, Magdalena Karwatka, Tracey Davey, Chris F. Inglehearn, Martin McKibbin, Reinhard Lührmann, David H. Steel, David J. Elliott, Lyle Armstrong, Henning Urlaub, Robin R. Ali, Sushma-Nagaraja Grellscheid, Colin A. Johnson, Sina Mozaffari-Jovin, and Majlinda Lako
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Science - Abstract
Abstract The carboxy-terminus of the spliceosomal protein PRPF8, which regulates the RNA helicase Brr2, is a hotspot for mutations causing retinitis pigmentosa-type 13, with unclear role in human splicing and tissue-specificity mechanism. We used patient induced pluripotent stem cells-derived cells, carrying the heterozygous PRPF8 c.6926 A > C (p.H2309P) mutation to demonstrate retinal-specific endophenotypes comprising photoreceptor loss, apical-basal polarity and ciliary defects. Comprehensive molecular, transcriptomic, and proteomic analyses revealed a role of the PRPF8/Brr2 regulation in 5’-splice site (5’SS) selection by spliceosomes, for which disruption impaired alternative splicing and weak/suboptimal 5’SS selection, and enhanced cryptic splicing, predominantly in ciliary and retinal-specific transcripts. Altered splicing efficiency, nuclear speckles organisation, and PRPF8 interaction with U6 snRNA, caused accumulation of active spliceosomes and poly(A)+ mRNAs in unique splicing clusters located at the nuclear periphery of photoreceptors. Collectively these elucidate the role of PRPF8/Brr2 regulatory mechanisms in splicing and the molecular basis of retinal disease, informing therapeutic approaches.
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- 2024
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9. The thymocyte-specific RNA-binding protein Arpp21 provides TCR repertoire diversity by binding to the 3’-UTR and promoting Rag1 mRNA expression
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Meng Xu, Taku Ito-Kureha, Hyun-Seo Kang, Aleksandar Chernev, Timsse Raj, Kai P. Hoefig, Christine Hohn, Florian Giesert, Yinhu Wang, Wenliang Pan, Natalia Ziętara, Tobias Straub, Regina Feederle, Carolin Daniel, Barbara Adler, Julian König, Stefan Feske, George C. Tsokos, Wolfgang Wurst, Henning Urlaub, Michael Sattler, Jan Kisielow, F. Gregory Wulczyn, Marcin Łyszkiewicz, and Vigo Heissmeyer
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Science - Abstract
Abstract The regulation of thymocyte development by RNA-binding proteins (RBPs) is largely unexplored. We identify 642 RBPs in the thymus and focus on Arpp21, which shows selective and dynamic expression in early thymocytes. Arpp21 is downregulated in response to T cell receptor (TCR) and Ca2+ signals. Downregulation requires Stim1/Stim2 and CaMK4 expression and involves Arpp21 protein phosphorylation, polyubiquitination and proteasomal degradation. Arpp21 directly binds RNA through its R3H domain, with a preference for uridine-rich motifs, promoting the expression of target mRNAs. Analysis of the Arpp21–bound transcriptome reveals strong interactions with the Rag1 3′-UTR. Arpp21–deficient thymocytes show reduced Rag1 expression, delayed TCR rearrangement and a less diverse TCR repertoire. This phenotype is recapitulated in Rag1 3′-UTR mutant mice harboring a deletion of the Arpp21 response region. These findings show how thymocyte-specific Arpp21 promotes Rag1 expression to enable TCR repertoire diversity until signals from the TCR terminate Arpp21 and Rag1 activities.
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- 2024
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10. Protein degradation by human 20S proteasomes elucidates the interplay between peptide hydrolysis and splicing
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Wai Tuck Soh, Hanna P. Roetschke, John A. Cormican, Bei Fang Teo, Nyet Cheng Chiam, Monika Raabe, Ralf Pflanz, Fabian Henneberg, Stefan Becker, Ashwin Chari, Haiyan Liu, Henning Urlaub, Juliane Liepe, and Michele Mishto
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Science - Abstract
Abstract If and how proteasomes catalyze not only peptide hydrolysis but also peptide splicing is an open question that has divided the scientific community. The debate has so far been based on immunopeptidomics, in vitro digestions of synthetic polypeptides as well as ex vivo and in vivo experiments, which could only indirectly describe proteasome-catalyzed peptide splicing of full-length proteins. Here we develop a workflow—and cognate software - to analyze proteasome-generated non-spliced and spliced peptides produced from entire proteins and apply it to in vitro digestions of 15 proteins, including well-known intrinsically disordered proteins such as human tau and α-Synuclein. The results confirm that 20S proteasomes produce a sizeable variety of cis-spliced peptides, whereas trans-spliced peptides are a minority. Both peptide hydrolysis and splicing produce peptides with well-defined characteristics, which hint toward an intricate regulation of both catalytic activities. At protein level, both non-spliced and spliced peptides are not randomly localized within protein sequences, but rather concentrated in hotspots of peptide products, in part driven by protein sequence motifs and proteasomal preferences. At sequence level, the different peptide sequence preference of peptide hydrolysis and peptide splicing suggests a competition between the two catalytic activities of 20S proteasomes during protein degradation.
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- 2024
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11. Prevalent and sex-biased breathing patterns modify functional connectivity MRI in young adults
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Jonathan D. Power, Marc J. Dubin, Rebecca M. Jones, Henning U. Voss, Alex Martin, Benjamin M. Silver, and Charles J. Lynch
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Adult ,Male ,0301 basic medicine ,medicine.medical_specialty ,genetic structures ,Science ,Rest ,General Physics and Astronomy ,Audiology ,behavioral disciplines and activities ,Brain mapping ,Article ,General Biochemistry, Genetics and Molecular Biology ,Young Adult ,03 medical and health sciences ,Sex Factors ,0302 clinical medicine ,Breathing pattern ,medicine ,Humans ,Young adult ,lcsh:Science ,Association (psychology) ,Brain Mapping ,Multidisciplinary ,medicine.diagnostic_test ,Resting state fMRI ,business.industry ,Respiration ,Brain ,Cognitive neuroscience ,Magnetic resonance imaging ,General Chemistry ,Human brain ,Magnetic Resonance Imaging ,030104 developmental biology ,medicine.anatomical_structure ,nervous system ,Breathing ,lcsh:Q ,Female ,business ,psychological phenomena and processes ,030217 neurology & neurosurgery - Abstract
Resting state functional connectivity magnetic resonance imaging (fMRI) is a tool for investigating human brain organization. Here we identify, visually and algorithmically, two prevalent influences on fMRI signals during 440 h of resting state scans in 440 healthy young adults, both caused by deviations from normal breathing which we term deep breaths and bursts. The two respiratory patterns have distinct influences on fMRI signals and signal covariance, distinct timescales, distinct cardiovascular correlates, and distinct tendencies to manifest by sex. Deep breaths are not sex-biased. Bursts, which are serial taperings of respiratory depth typically spanning minutes at a time, are more common in males. Bursts share features of chemoreflex-driven clinical breathing patterns that also occur primarily in males, with notable neurological, psychiatric, medical, and lifespan associations. These results identify common breathing patterns in healthy young adults with distinct influences on functional connectivity and an ability to differentially influence resting state fMRI studies., Functional connectivity measured from fMRI data is widely used in neuroscience. Here the authors report an association between two types of breathing signature and obtained BOLD data, and associated sex differences.
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- 2020
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12. Control of intracellular pH and bicarbonate by CO2 diffusion into human sperm
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Elena Grahn, Svenja V. Kaufmann, Malika Askarova, Momchil Ninov, Luisa M. Welp, Thomas K. Berger, Henning Urlaub, and U.Benjamin Kaupp
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Science - Abstract
Abstract The reaction of CO2 with H2O to form bicarbonate (HCO3 −) and H+ controls sperm motility and fertilization via HCO3 −-stimulated cAMP synthesis. A complex network of signaling proteins participates in this reaction. Here, we identify key players that regulate intracellular pH (pHi) and HCO3 − in human sperm by quantitative mass spectrometry (MS) and kinetic patch-clamp fluorometry. The resting pHi is set by amiloride-sensitive Na+/H+ exchange. The sperm-specific putative Na+/H+ exchanger SLC9C1, unlike its sea urchin homologue, is not gated by voltage or cAMP. Transporters and channels implied in HCO3 − transport are not detected, and may be present at copy numbers
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- 2023
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13. Extended DNA threading through a dual-engine motor module of the activating signal co-integrator 1 complex
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Junqiao Jia, Tarek Hilal, Katherine E. Bohnsack, Aleksandar Chernev, Ning Tsao, Juliane Bethmann, Aruna Arumugam, Lane Parmely, Nicole Holton, Bernhard Loll, Nima Mosammaparast, Markus T. Bohnsack, Henning Urlaub, and Markus C. Wahl
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Science - Abstract
ASCC3 is a multi-functional helicase that contains two consecutive Ski2-like helicase units. Here, the authors show that ASCC3 can unwind DNA by threading one strand of a substrate duplex through both helicase units, supported by the TRIP4 protein.
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- 2023
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14. Sexual dimorphism in striatal dopaminergic responses promotes monogamy in social songbirds
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Paresh J. Kothari, Ofer Tchernichovski, Santosh A. Helekar, Henning U. Voss, Iva Ljubičić, Julia Hyland Bruno, and Kirill Tokarev
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Male ,0301 basic medicine ,Statistics as Topic ,Synaptic Transmission ,Sexual Behavior, Animal ,0302 clinical medicine ,Biology (General) ,Sex Characteristics ,Ecology ,General Neuroscience ,Dopaminergic ,General Medicine ,reward circuitry ,Sexual behavior ,Head Movements ,behavior and behavior mechanisms ,Medicine ,Female ,Brain stimulation reward ,dopamine ,Reinforcement, Psychology ,psychological phenomena and processes ,Research Article ,animal structures ,QH301-705.5 ,Movement ,Science ,Zoology ,Biology ,Stimulus (physiology) ,General Biochemistry, Genetics and Molecular Biology ,social behavior ,03 medical and health sciences ,monogamy ,Animals ,Zebra finch ,General Immunology and Microbiology ,Receptors, Dopamine D2 ,zebra finch ,biology.organism_classification ,Mating system ,Songbird ,Neostriatum ,Sexual dimorphism ,030104 developmental biology ,Acoustic Stimulation ,nervous system ,sexual dimorphism ,Other ,Finches ,Vocalization, Animal ,030217 neurology & neurosurgery ,Neuroscience - Abstract
In many songbird species, males sing to attract females and repel rivals. How can gregarious, non-territorial songbirds such as zebra finches, where females have access to numerous males, sustain monogamy? We found that the dopaminergic reward circuitry of zebra finches can simultaneously promote social cohesion and breeding boundaries. Surprisingly, in unmated males but not in females, striatal dopamine neurotransmission was elevated after hearing songs. Behaviorally too, unmated males but not females persistently exchanged mild punishments in return for songs. Song reinforcement diminished when dopamine receptors were blocked. In females, we observed song reinforcement exclusively to the mate’s song, although their striatal dopamine neurotransmission was only slightly elevated. These findings suggest that song-triggered dopaminergic activation serves a dual function in social songbirds: as low-threshold social reinforcement in males and as ultra-selective sexual reinforcement in females. Co-evolution of sexually dimorphic reinforcement systems can explain the coexistence of gregariousness and monogamy., eLife digest While monogamy is rare within the animal kingdom, some species – including humans and many birds – can be highly social and yet sustain monogamous relationships. Zebra finches, for example, are among a number of species of songbirds in which numerous males and females live closely together but maintain monogamous partnerships. Male songbirds use their songs to attract females, who do not themselves sing. But if female birds are attracted to any male song, how do they manage to remain monogamous when surrounded by potential suitors? In songbirds, and in humans too, a region of the brain called the striatum regulates both social and sexual behaviors. It does this by modulating the release of a molecule called dopamine, which is the brain’s reward signal. Tokarev et al. show that hearing songs triggers dopamine release within the striatum of unattached male zebra finches, but has no such effect in single females. Unattached male songbirds will also put up with irritating puffs of air in exchange for being able to watch videos of singing birds, whereas unattached females will not. Female songbirds with partners will tolerate the air puffs, but only if the videos are accompanied with the songs of their own mate. These findings suggest that song serves as a generic social stimulus for zebra finch males, helping large numbers of birds to live together. By contrast, for a female zebra finch, the song of her partner is a highly selective sexual stimulus. These sex-specific responses to the same socially-relevant stimuli may explain how gregarious animals are able to maintain monogamous pair bonds. More generally, these results are a step towards understanding how brain reward systems regulate social interactions. Studying these mechanisms in songbird species with different social and mating systems could ultimately provide insights into social and sexual behavior in people.
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- 2017
15. 5-HT-dependent synaptic plasticity of the prefrontal cortex in postnatal development
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Guilherme Shigueto Vilar Higa, José Francis-Oliveira, Estevão Carlos-Lima, Alicia Moraes Tamais, Fernando da Silva Borges, Alexandre Hiroaki Kihara, Ianê Carvalho Shieh, Henning Ulrich, Silvana Chiavegatto, and Roberto De Pasquale
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Medicine ,Science - Abstract
Abstract Important functions of the prefrontal cortex (PFC) are established during early life, when neurons exhibit enhanced synaptic plasticity and synaptogenesis. This developmental stage drives the organization of cortical connectivity, responsible for establishing behavioral patterns. Serotonin (5-HT) emerges among the most significant factors that modulate brain activity during postnatal development. In the PFC, activated 5-HT receptors modify neuronal excitability and interact with intracellular signaling involved in synaptic modifications, thus suggesting that 5-HT might participate in early postnatal plasticity. To test this hypothesis, we employed intracellular electrophysiological recordings of PFC layer 5 neurons to study the modulatory effects of 5-HT on plasticity induced by theta-burst stimulation (TBS) in two postnatal periods of rats. Our results indicate that 5-HT is essential for TBS to result in synaptic changes during the third postnatal week, but not later. TBS coupled with 5-HT2A or 5-HT1A and 5-HT7 receptors stimulation leads to long-term depression (LTD). On the other hand, TBS and synergic activation of 5-HT1A, 5-HT2A, and 5-HT7 receptors lead to long-term potentiation (LTP). Finally, we also show that 5-HT dependent synaptic plasticity of the PFC is impaired in animals that are exposed to early-life chronic stress.
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- 2022
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16. Calpain cleavage of Junctophilin-2 generates a spectrum of calcium-dependent cleavage products and DNA-rich NT1-fragment domains in cardiomyocytes
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Gunnar Weninger, Tatiana Pochechueva, Dana El Chami, Xiaojing Luo, Tobias Kohl, Sören Brandenburg, Henning Urlaub, Kaomei Guan, Christof Lenz, and Stephan E. Lehnart
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Medicine ,Science - Abstract
Abstract Calpains are calcium-activated neutral proteases involved in the regulation of key signaling pathways. Junctophilin-2 (JP2) is a Calpain-specific proteolytic target and essential structural protein inside Ca2+ release units required for excitation-contraction coupling in cardiomyocytes. While downregulation of JP2 by Calpain cleavage in heart failure has been reported, the precise molecular identity of the Calpain cleavage sites and the (patho-)physiological roles of the JP2 proteolytic products remain controversial. We systematically analyzed the JP2 cleavage fragments as function of Calpain-1 versus Calpain-2 proteolytic activities, revealing that both Calpain isoforms preferentially cleave mouse JP2 at R565, but subsequently at three additional secondary Calpain cleavage sites. Moreover, we identified the Calpain-specific primary cleavage products for the first time in human iPSC-derived cardiomyocytes. Knockout of RyR2 in hiPSC-cardiomyocytes destabilized JP2 resulting in an increase of the Calpain-specific cleavage fragments. The primary N-terminal cleavage product NT1 accumulated in the nucleus of mouse and human cardiomyocytes in a Ca2+-dependent manner, closely associated with euchromatic chromosomal regions, where NT1 is proposed to function as a cardio-protective transcriptional regulator in heart failure. Taken together, our data suggest that stabilizing NT1 by preventing secondary cleavage events by Calpain and other proteases could be an important therapeutic target for future studies.
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- 2022
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17. Hsp multichaperone complex buffers pathologically modified Tau
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Antonia Moll, Lisa Marie Ramirez, Momchil Ninov, Juliane Schwarz, Henning Urlaub, and Markus Zweckstetter
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Science - Abstract
Alzheimer’s disease is characterized by the accumulation of aggregated tau protein. Here the authors find that Hsp chaperones, which normally protect cell homeostasis, can assemble with co-chaperones in a “multichaperone machinery” to target tau aggregation.
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- 2022
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18. Identification of distinct cytotoxic granules as the origin of supramolecular attack particles in T lymphocytes
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Hsin-Fang Chang, Claudia Schirra, Momchil Ninov, Ulrike Hahn, Keerthana Ravichandran, Elmar Krause, Ute Becherer, Štefan Bálint, Maria Harkiolaki, Henning Urlaub, Salvatore Valitutti, Cosima T. Baldari, Michael L. Dustin, Reinhard Jahn, and Jens Rettig
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Science - Abstract
Cytotoxic T cells have specialised granules that are important for mediating their killing function. Here the authors characterise two types of cytotoxic granules and indicate different functions and temporal release of mediators at the immunological synapse.
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- 2022
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19. Proteomic mapping of atrial and ventricular heart tissue in patients with aortic valve stenosis
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Boris Barbarics, Katja Eildermann, Lars Kaderali, Lukas Cyganek, Uwe Plessmann, Julius Bodemeyer, Thomas Paul, Philipp Ströbel, Henning Urlaub, Theodorus Tirilomis, Christof Lenz, and Hanibal Bohnenberger
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Medicine ,Science - Abstract
Abstract Aortic valve stenosis (AVS) is one of the most common valve diseases in the world. However, detailed biological understanding of the myocardial changes in AVS hearts on the proteome level is still lacking. Proteomic studies using high-resolution mass spectrometry of formalin-fixed and paraffin-embedded (FFPE) human myocardial tissue of AVS-patients are very rare due to methodical issues. To overcome these issues this study used high resolution mass spectrometry in combination with a stem cell-derived cardiac specific protein quantification-standard to profile the proteomes of 17 atrial and 29 left ventricular myocardial FFPE human myocardial tissue samples from AVS-patients. In our proteomic analysis we quantified a median of 1980 (range 1495–2281) proteins in every single sample and identified significant upregulation of 239 proteins in atrial and 54 proteins in ventricular myocardium. We compared the proteins with published data. Well studied proteins reflect disease-related changes in AVS, such as cardiac hypertrophy, development of fibrosis, impairment of mitochondria and downregulated blood supply. In summary, we provide both a workflow for quantitative proteomics of human FFPE heart tissue and a comprehensive proteomic resource for AVS induced changes in the human myocardium.
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- 2021
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20. The RNA helicase Dbp7 promotes domain V/VI compaction and stabilization of inter-domain interactions during early 60S assembly
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Gerald Ryan R. Aquino, Philipp Hackert, Nicolai Krogh, Kuan-Ting Pan, Mariam Jaafar, Anthony K. Henras, Henrik Nielsen, Henning Urlaub, Katherine E. Bohnsack, and Markus T. Bohnsack
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Science - Abstract
Early steps of large 60S ribosomal subunit biogenesis are not well understood. Here, the authors combine biochemical experiments with protein-RNA crosslinking and mass spectrometry to show that the RNA helicase Dbp7 is key player during early 60S ribosomal assembly. Dbp7 regulates a series of events driving compaction of domain V/VI in early pre60S ribosomal particles.
- Published
- 2021
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21. Mapping protein interactions in the active TOM-TIM23 supercomplex
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Ridhima Gomkale, Andreas Linden, Piotr Neumann, Alexander Benjamin Schendzielorz, Stefan Stoldt, Olexandr Dybkov, Markus Kilisch, Christian Schulz, Luis Daniel Cruz-Zaragoza, Blanche Schwappach, Ralf Ficner, Stefan Jakobs, Henning Urlaub, and Peter Rehling
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Science - Abstract
The TOM and TIM23 complexes facilitate the transport of nuclear-encoded proteins into the mitochondrial matrix. Here, the authors use a stalled client protein to purify the translocation supercomplex and gain insight into the TOM-TIM23 interface and the mechanism of protein handover from the TOM to the TIM23 complex.
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- 2021
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22. Structural basis of GTPase-mediated mitochondrial ribosome biogenesis and recycling
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Hauke S. Hillen, Elena Lavdovskaia, Franziska Nadler, Elisa Hanitsch, Andreas Linden, Katherine E. Bohnsack, Henning Urlaub, and Ricarda Richter-Dennerlein
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Science - Abstract
Maturation of the ribosomal peptidyl transferase center (PTC) is mediated by universally conserved GTPases. Here, cryo-EM structures of mitochondrial ribosomal large subunit assembly intermediates and of mature ribosomes offer insight into the roles of several assembly factors, including GTPBP6’s role in both ribosome biogenesis and recycling.
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- 2021
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23. Translation error clusters induced by aminoglycoside antibiotics
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Ingo Wohlgemuth, Raffaella Garofalo, Ekaterina Samatova, Aybeg Nafiz Günenç, Christof Lenz, Henning Urlaub, and Marina V. Rodnina
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Science - Abstract
Aminoglycoside antibiotics target the ribosome and induce misreading, yet which translation errors induce bacterial cell death is unclear. Here authors use quantitative mass spectrometry and show that bactericidal aminoglycosides induce clusters of errors in full-length proteins in vivo with as many as four amino acid substitutions in a row.
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- 2021
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24. Analysis of protein-DNA interactions in chromatin by UV induced cross-linking and mass spectrometry
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Alexandra Stützer, Luisa M. Welp, Monika Raabe, Timo Sachsenberg, Christin Kappert, Alexander Wulf, Andy M. Lau, Stefan-Sebastian David, Aleksandar Chernev, Katharina Kramer, Argyris Politis, Oliver Kohlbacher, Wolfgang Fischle, and Henning Urlaub
- Subjects
Science - Abstract
Cross-linking mass spectrometry (XLMS) allows mapping of protein-protein and protein-RNA interactions, but the analysis of protein-DNA complexes remains challenging. Here, the authors develop a UV light-based XLMS workflow to determine protein-DNA interfaces in reconstituted chromatin and isolated nuclei.
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- 2020
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25. A streamlined pipeline for multiplexed quantitative site-specific N-glycoproteomics
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Pan Fang, Yanlong Ji, Ivan Silbern, Carmen Doebele, Momchil Ninov, Christof Lenz, Thomas Oellerich, Kuan-Ting Pan, and Henning Urlaub
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Science - Abstract
Comprehensive quantitative profiling of intact glycopeptides remains technically challenging. To address this, the authors here develop an integrated quantitative glycoproteomic workflow, including optimized sample preparation, multiplexed quantification and a dedicated data processing tool.
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- 2020
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26. The Archaeal Proteome Project advances knowledge about archaeal cell biology through comprehensive proteomics
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Stefan Schulze, Zachary Adams, Micaela Cerletti, Rosana De Castro, Sébastien Ferreira-Cerca, Christian Fufezan, María Inés Giménez, Michael Hippler, Zivojin Jevtic, Robert Knüppel, Georgio Legerme, Christof Lenz, Anita Marchfelder, Julie Maupin-Furlow, Roberto A. Paggi, Friedhelm Pfeiffer, Ansgar Poetsch, Henning Urlaub, and Mechthild Pohlschroder
- Subjects
Science - Abstract
While archaeal proteomics advanced rapidly, a comprehensive proteome database for archaea is lacking. Therefore, the authors here launch the Archaeal Proteome Project, a community-effort providing insights into archaeal cell biology via the combined reanalysis of Haloferax volcanii proteomics data.
- Published
- 2020
- Full Text
- View/download PDF
27. Receptor-specific Ca2+ oscillation patterns mediated by differential regulation of P2Y purinergic receptors in rat hepatocytes
- Author
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Juliana C. Corrêa-Velloso, Paula J. Bartlett, Robert Brumer, Lawrence D. Gaspers, Henning Ulrich, and Andrew P. Thomas
- Subjects
Biological sciences ,Cell biology ,Cellular physiology ,Functional aspects of cell biology ,Science - Abstract
Summary: Extracellular agonists linked to inositol-1,4,5-trisphosphate (IP3) formation elicit cytosolic Ca2+ oscillations in many cell types, but despite a common signaling pathway, distinct agonist-specific Ca2+ spike patterns are observed. Using qPCR, we show that rat hepatocytes express multiple purinergic P2Y and P2X receptors (R). ADP acting through P2Y1R elicits narrow Ca2+ oscillations, whereas UTP acting through P2Y2R elicits broad Ca2+ oscillations, with composite patterns observed for ATP. P2XRs do not play a role at physiological agonist levels. The discrete Ca2+ signatures reflect differential effects of protein kinase C (PKC), which selectively modifies the falling phase of the Ca2+ spikes. Negative feedback by PKC limits the duration of P2Y1R-induced Ca2+ spikes in a manner that requires extracellular Ca2+. By contrast, P2Y2R is resistant to PKC negative feedback. Thus, the PKC leg of the bifurcated IP3 signaling pathway shapes unique Ca2+ oscillation patterns that allows for distinct cellular responses to different agonists.
- Published
- 2021
- Full Text
- View/download PDF
28. Therapeutic Effects of Cannabinoids and Their Applications in COVID-19 Treatment
- Author
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Rebeca Pérez, Talita Glaser, Cecilia Villegas, Viviana Burgos, Henning Ulrich, and Cristian Paz
- Subjects
refractory epilepsy ,COVID-19 ,phytocannabinoids ,synthetic cannabinoids ,clinical trials ,Science - Abstract
Cannabis sativa is one of the first medicinal plants used by humans. Its medical use remains controversial because it is a psychotropic drug whose use has been banned. Recently, however, some countries have approved its use, including for recreational and medical purposes, and have allowed the scientific study of its compounds. Cannabis is characterized by the production of special types of natural products called phytocannabinoids that are synthesized exclusively by this genus. Phytocannabinoids and endocannabinoids are chemically different, but both pharmacologically modulate CB1, CB2, GRP55, GRP119 and TRPV1 receptor activities, involving activities such as memory, sleep, mood, appetite and motor regulation, pain sensation, neuroinflammation, neurogenesis and apoptosis. Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD) are phytocannabinoids with greater pharmacological potential, including anti-inflammatory, neuroprotective and anticonvulsant activities. Cannabidiol is showing promising results for the treatment of COVID-19, due to its capability of acting on the unleashed cytokine storm, on the proteins necessary for both virus entry and replication and on the neurological consequences of patients who have been infected by the virus. Here, we summarize the latest knowledge regarding the advantages of using cannabinoids in the treatment of COVID-19.
- Published
- 2022
- Full Text
- View/download PDF
29. Insights into the assembly and architecture of a Staufen-mediated mRNA decay (SMD)-competent mRNP
- Author
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Manjeera Gowravaram, Juliane Schwarz, Sana K. Khilji, Henning Urlaub, and Sutapa Chakrabarti
- Subjects
Science - Abstract
The Staufen proteins recognize secondary structures in 3’-untranslated regions in mRNA transcripts and induce degradation of these mRNAs with the help of the RNA helicase UPF1. Here the authors report that the nonsense-mediated mRNA decay factor UPF2 mediates the interaction between Stau1 and UPF1 in Staufen-mediated mRNA decay.
- Published
- 2019
- Full Text
- View/download PDF
30. Structural basis of TFIIH activation for nucleotide excision repair
- Author
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Goran Kokic, Aleksandar Chernev, Dimitry Tegunov, Christian Dienemann, Henning Urlaub, and Patrick Cramer
- Subjects
Science - Abstract
The NER machinery contains the multisubunit transcription factor IIH (TFIIH) that opens the DNA repair bubble, scans for the lesion, and coordinates excision of the damaged site. Here the authors resolve the cryo-electron microscopy structure of the human core TFIIH-XPA-DNA complex and provide insights into its activation.
- Published
- 2019
- Full Text
- View/download PDF
31. Exocyst-mediated membrane trafficking of the lissencephaly-associated ECM receptor dystroglycan is required for proper brain compartmentalization
- Author
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Andriy S Yatsenko, Mariya M Kucherenko, Yuanbin Xie, Henning Urlaub, and Halyna R Shcherbata
- Subjects
dystroglycan ,cobblestone lissencephaly ,exocyst ,differentiating neurons ,neuronal Dg interactome ,brain neuropils ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
To assemble a brain, differentiating neurons must make proper connections and establish specialized brain compartments. Abnormal levels of cell adhesion molecules disrupt these processes. Dystroglycan (Dg) is a major non-integrin cell adhesion receptor, deregulation of which is associated with dramatic neuroanatomical defects such as lissencephaly type II or cobblestone brain. The previously established Drosophila model for cobblestone lissencephaly was used to understand how Dg is regulated in the brain. During development, Dg has a spatiotemporally dynamic expression pattern, fine-tuning of which is crucial for accurate brain assembly. In addition, mass spectrometry analyses identified numerous components associated with Dg in neurons, including several proteins of the exocyst complex. Data show that exocyst-based membrane trafficking of Dg allows its distinct expression pattern, essential for proper brain morphogenesis. Further studies of the Dg neuronal interactome will allow identification of new factors involved in the development of dystroglycanopathies and advance disease diagnostics in humans.
- Published
- 2021
- Full Text
- View/download PDF
32. RNA helicases mediate structural transitions and compositional changes in pre-ribosomal complexes
- Author
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Lukas Brüning, Philipp Hackert, Roman Martin, Jimena Davila Gallesio, Gerald Ryan R. Aquino, Henning Urlaub, Katherine E. Sloan, and Markus T. Bohnsack
- Subjects
Science - Abstract
Pre-ribosomes undergo numerous structural rearrangements during their assembly. Here the authors identify the binding sites of three essential RNA helicases on pre-ribosomal particles, enabling them to provide insights into the structural and compositional changes that occur during biogenesis of the large ribosomal subunit.
- Published
- 2018
- Full Text
- View/download PDF
33. Disrupted alternative splicing for genes implicated in splicing and ciliogenesis causes PRPF31 retinitis pigmentosa
- Author
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Adriana Buskin, Lili Zhu, Valeria Chichagova, Basudha Basu, Sina Mozaffari-Jovin, David Dolan, Alastair Droop, Joseph Collin, Revital Bronstein, Sudeep Mehrotra, Michael Farkas, Gerrit Hilgen, Kathryn White, Kuan-Ting Pan, Achim Treumann, Dean Hallam, Katarzyna Bialas, Git Chung, Carla Mellough, Yuchun Ding, Natalio Krasnogor, Stefan Przyborski, Simon Zwolinski, Jumana Al-Aama, Sameer Alharthi, Yaobo Xu, Gabrielle Wheway, Katarzyna Szymanska, Martin McKibbin, Chris F. Inglehearn, David J. Elliott, Susan Lindsay, Robin R. Ali, David H. Steel, Lyle Armstrong, Evelyne Sernagor, Henning Urlaub, Eric Pierce, Reinhard Lührmann, Sushma-Nagaraja Grellscheid, Colin A. Johnson, and Majlinda Lako
- Subjects
Science - Abstract
Mutations in pre-mRNA processing factors cause autosomal dominant retinitis pigmentosa. Here the authors provide insights into the pathophysiological mechanisms underlying non-syndromic retinal disease caused by heterozygous mutations in genes encoding ubiquitously expressed splicing factors.
- Published
- 2018
- Full Text
- View/download PDF
34. Precisely measured protein lifetimes in the mouse brain reveal differences across tissues and subcellular fractions
- Author
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Eugenio F. Fornasiero, Sunit Mandad, Hanna Wildhagen, Mihai Alevra, Burkhard Rammner, Sarva Keihani, Felipe Opazo, Inga Urban, Till Ischebeck, M. Sadman Sakib, Maryam K. Fard, Koray Kirli, Tonatiuh Pena Centeno, Ramon O. Vidal, Raza-Ur Rahman, Eva Benito, André Fischer, Sven Dennerlein, Peter Rehling, Ivo Feussner, Stefan Bonn, Mikael Simons, Henning Urlaub, and Silvio O. Rizzoli
- Subjects
Science - Abstract
Measuring precise protein turnover rates in animals is technically challenging at the proteomic level. Here, Fornasiero and colleagues use isotopic labeling with mass spectrometry and mathematical modeling to accurately determine protein lifetimes in the mouse brain
- Published
- 2018
- Full Text
- View/download PDF
35. Structure and pro-toxic mechanism of the human Hsp90/PPIase/Tau complex
- Author
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Javier Oroz, Bliss J. Chang, Piotr Wysoczanski, Chung-Tien Lee, Ángel Pérez-Lara, Pijush Chakraborty, Romina V. Hofele, Jeremy D. Baker, Laura J. Blair, Jacek Biernat, Henning Urlaub, Eckhard Mandelkow, Chad A. Dickey, and Markus Zweckstetter
- Subjects
Science - Abstract
The chaperone Hsp90 plays a key role in maintaining cellular homeostasis. Here the authors provide structural insights into substrate recognition and the pro-folding mechanism of Hsp90/co-chaperone complexes by studying the complex of Hsp90 with its co-chaperone FKBP51 and the substrate Tau bound Hsp90/FKBP51 ternary complex using a NMR based integrative approach.
- Published
- 2018
- Full Text
- View/download PDF
36. Architecture of the RNA polymerase II-Paf1C-TFIIS transcription elongation complex
- Author
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Youwei Xu, Carrie Bernecky, Chung-Tien Lee, Kerstin C. Maier, Björn Schwalb, Dimitry Tegunov, Jürgen M. Plitzko, Henning Urlaub, and Patrick Cramer
- Subjects
Science - Abstract
The Paf1 complex (Paf1C) is an elongation factor assembly that forms the interface between transcribing Pol II and chromatin factors. Here the authors describe the architecture of Paf1C and its interface with Pol II, and show that Paf1C is globally required for normal mRNA transcription in yeast.
- Published
- 2017
- Full Text
- View/download PDF
37. Increased expression of heme-binding protein 1 early in Alzheimer's disease is linked to neurotoxicity
- Author
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Oleksandr Yagensky, Mahdokht Kohansal-Nodehi, Saravanan Gunaseelan, Tamara Rabe, Saima Zafar, Inga Zerr, Wolfgang Härtig, Henning Urlaub, and John JE Chua
- Subjects
neuron ,Alzheimer's disease ,Hebp1 ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Alzheimer’s disease is the most prevalent neurodegenerative disorder leading to progressive cognitive decline. Despite decades of research, understanding AD progression at the molecular level, especially at its early stages, remains elusive. Here, we identified several presymptomatic AD markers by investigating brain proteome changes over the course of neurodegeneration in a transgenic mouse model of AD (3×Tg-AD). We show that one of these markers, heme-binding protein 1 (Hebp1), is elevated in the brains of both 3×Tg-AD mice and patients affected by rapidly-progressing forms of AD. Hebp1, predominantly expressed in neurons, interacts with the mitochondrial contact site complex (MICOS) and exhibits a perimitochondrial localization. Strikingly, wildtype, but not Hebp1-deficient, neurons showed elevated cytotoxicity in response to heme-induced apoptosis. Increased survivability in Hebp1-deficient neurons is conferred by blocking the activation of the mitochondrial-associated caspase signaling pathway. Taken together, our data highlight a role of Hebp1 in progressive neuronal loss during AD progression.
- Published
- 2019
- Full Text
- View/download PDF
38. A spliceosome intermediate with loosely associated tri-snRNP accumulates in the absence of Prp28 ATPase activity
- Author
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Carsten Boesler, Norbert Rigo, Maria M. Anokhina, Marcel J. Tauchert, Dmitry E. Agafonov, Berthold Kastner, Henning Urlaub, Ralf Ficner, Cindy L. Will, and Reinhard Lührmann
- Subjects
Science - Abstract
The assembly of the splicesome involves several distinct stages that require the sequential action of DExD/H-box RNA helicases. Here, the authors uncover a new intermediate, the pre-B complex, that accumulates in the presence of an inactive form of the DEAD-box protein Prp28.
- Published
- 2016
- Full Text
- View/download PDF
39. Dynamic and flexible H3K9me3 bridging via HP1β dimerization establishes a plastic state of condensed chromatin
- Author
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Kyoko Hiragami-Hamada, Szabolcs Soeroes, Miroslav Nikolov, Bryan Wilkins, Sarah Kreuz, Carol Chen, Inti A. De La Rosa-Velázquez, Hans Michael Zenn, Nils Kost, Wiebke Pohl, Aleksandar Chernev, Dirk Schwarzer, Thomas Jenuwein, Matthew Lorincz, Bastian Zimmermann, Peter Jomo Walla, Heinz Neumann, Tuncay Baubec, Henning Urlaub, and Wolfgang Fischle
- Subjects
Science - Abstract
Heterochromatin protein 1 (HP1), including HP1 α, β and γ, is a family of non-histone chromatin factors thought to be involved in chromatin organization. Here, the authors show that dimeric HP1β interacts dynamically with H3K9me3, a hallmark of heterochromatin, and bridges condensed chromatin.
- Published
- 2016
- Full Text
- View/download PDF
40. Identification of a small molecule inhibitor that stalls splicing at an early step of spliceosome activation
- Author
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Anzhalika Sidarovich, Cindy L Will, Maria M Anokhina, Javier Ceballos, Sonja Sievers, Dmitry E Agafonov, Timur Samatov, Penghui Bao, Berthold Kastner, Henning Urlaub, Herbert Waldmann, and Reinhard Lührmann
- Subjects
spliceosome ,pre-mRNA splicing ,small molecule inhibitor ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Small molecule inhibitors of pre-mRNA splicing are important tools for identifying new spliceosome assembly intermediates, allowing a finer dissection of spliceosome dynamics and function. Here, we identified a small molecule that inhibits human pre-mRNA splicing at an intermediate stage during conversion of pre-catalytic spliceosomal B complexes into activated Bact complexes. Characterization of the stalled complexes (designated B028) revealed that U4/U6 snRNP proteins are released during activation before the U6 Lsm and B-specific proteins, and before recruitment and/or stable incorporation of Prp19/CDC5L complex and other Bact complex proteins. The U2/U6 RNA network in B028 complexes differs from that of the Bact complex, consistent with the idea that the catalytic RNA core forms stepwise during the B to Bact transition and is likely stabilized by the Prp19/CDC5L complex and related proteins. Taken together, our data provide new insights into the RNP rearrangements and extensive exchange of proteins that occurs during spliceosome activation.
- Published
- 2017
- Full Text
- View/download PDF
41. DPP9 is a novel component of the N-end rule pathway targeting the tyrosine kinase Syk
- Author
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Daniela Justa-Schuch, Maria Silva-Garcia, Esther Pilla, Michael Engelke, Markus Kilisch, Christof Lenz, Ulrike Möller, Fumihiko Nakamura, Henning Urlaub, and Ruth Geiss-Friedlander
- Subjects
Syk ,DPP9 ,N-end rule ,protein half-life ,Cbl ,B cell signalling ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
The aminopeptidase DPP9 removes dipeptides from N-termini of substrates having a proline or alanine in second position. Although linked to several pathways including cell survival and metabolism, the molecular mechanisms underlying these outcomes are poorly understood. We identified a novel interaction of DPP9 with Filamin A, which recruits DPP9 to Syk, a central kinase in B-cell signalling. Syk signalling can be terminated by degradation, requiring the ubiquitin E3 ligase Cbl. We show that DPP9 cleaves Syk to produce a neo N-terminus with serine in position 1. Pulse-chases combined with mutagenesis studies reveal that Ser1 strongly influences Syk stability. Furthermore, DPP9 silencing reduces Cbl interaction with Syk, suggesting that DPP9 processing is a prerequisite for Syk ubiquitination. Consistently, DPP9 inhibition stabilizes Syk, thereby modulating Syk signalling. Taken together, we demonstrate DPP9 as a negative regulator of Syk and conclude that DPP9 is a novel integral aminopeptidase of the N-end rule pathway.
- Published
- 2016
- Full Text
- View/download PDF
42. Analysis of protein phosphorylation in nerve terminal reveals extensive changes in active zone proteins upon exocytosis
- Author
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Mahdokht Kohansal-Nodehi, John JE Chua, Henning Urlaub, Reinhard Jahn, and Dominika Czernik
- Subjects
phosphorylation ,synaptosome ,phosphoproteomics ,exocytosis ,active zone ,protein kinase ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Neurotransmitter release is mediated by the fast, calcium-triggered fusion of synaptic vesicles with the presynaptic plasma membrane, followed by endocytosis and recycling of the membrane of synaptic vesicles. While many of the proteins governing these processes are known, their regulation is only beginning to be understood. Here we have applied quantitative phosphoproteomics to identify changes in phosphorylation status of presynaptic proteins in resting and stimulated nerve terminals isolated from the brains of Wistar rats. Using rigorous quantification, we identified 252 phosphosites that are either up- or downregulated upon triggering calcium-dependent exocytosis. Particularly pronounced were regulated changes of phosphosites within protein constituents of the presynaptic active zone, including bassoon, piccolo, and RIM1. Additionally, we have mapped kinases and phosphatases that are activated upon stimulation. Overall, our study provides a snapshot of phosphorylation changes associated with presynaptic activity and provides a foundation for further functional analysis of key phosphosites involved in presynaptic plasticity.
- Published
- 2016
- Full Text
- View/download PDF
43. The target of the DEAH-box NTP triphosphatase Prp43 in Saccharomyces cerevisiae spliceosomes is the U2 snRNP-intron interaction
- Author
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Jean-Baptiste Fourmann, Olexandr Dybkov, Dmitry E Agafonov, Marcel J Tauchert, Henning Urlaub, Ralf Ficner, Patrizia Fabrizio, and Reinhard Lührmann
- Subjects
Human ,spliceosome ,helicases ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
The DEAH-box NTPase Prp43 and its cofactors Ntr1 and Ntr2 form the NTR complex and are required for disassembling intron-lariat spliceosomes (ILS) and defective earlier spliceosomes. However, the Prp43 binding site in the spliceosome and its target(s) are unknown. We show that Prp43 fused to Ntr1's G-patch motif (Prp43_Ntr1GP) is as efficient as the NTR in ILS disassembly, yielding identical dissociation products and recognizing its natural ILS target even in the absence of Ntr1’s C-terminal-domain (CTD) and Ntr2. Unlike the NTR, Prp43_Ntr1GP disassembles earlier spliceosomal complexes (A, B, Bact), indicating that Ntr2/Ntr1-CTD prevents NTR from disrupting properly assembled spliceosomes other than the ILS. The U2 snRNP-intron interaction is disrupted in all complexes by Prp43_Ntr1GP, and in the spliceosome contacts U2 proteins and the pre-mRNA, indicating that the U2 snRNP-intron interaction is Prp43’s major target.
- Published
- 2016
- Full Text
- View/download PDF
44. Correction: Nanobodies: site-specific labeling for super-resolution imaging, rapid epitope-mapping and native protein complex isolation
- Author
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Tino Pleiner, Mark Bates, Sergei Trakhanov, Chung-Tien Lee, Jan Erik Schliep, Hema Chug, Marc Böhning, Holger Stark, Henning Urlaub, and Dirk Görlich
- Subjects
Medicine ,Science ,Biology (General) ,QH301-705.5 - Published
- 2016
- Full Text
- View/download PDF
45. Nanobodies: site-specific labeling for super-resolution imaging, rapid epitope-mapping and native protein complex isolation
- Author
-
Tino Pleiner, Mark Bates, Sergei Trakhanov, Chung-Tien Lee, Jan Erik Schliep, Hema Chug, Marc Böhning, Holger Stark, Henning Urlaub, and Dirk Görlich
- Subjects
nuclear pore complex ,nanobody ,label displacement ,site-specific ,native purification ,epitope mapping ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Nanobodies are single-domain antibodies of camelid origin. We generated nanobodies against the vertebrate nuclear pore complex (NPC) and used them in STORM imaging to locate individual NPC proteins with
- Published
- 2015
- Full Text
- View/download PDF
46. A deep proteomics perspective on CRM1-mediated nuclear export and nucleocytoplasmic partitioning
- Author
-
Koray Kırlı, Samir Karaca, Heinz Jürgen Dehne, Matthias Samwer, Kuan Ting Pan, Christof Lenz, Henning Urlaub, and Dirk Görlich
- Subjects
XPO1/CRM1 ,NES ,protein localization ,protein transport ,exportin ,Nup ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
CRM1 is a highly conserved, RanGTPase-driven exportin that carries proteins and RNPs from the nucleus to the cytoplasm. We now explored the cargo-spectrum of CRM1 in depth and identified surprisingly large numbers, namely >700 export substrates from the yeast S. cerevisiae, ≈1000 from Xenopus oocytes and >1050 from human cells. In addition, we quantified the partitioning of ≈5000 unique proteins between nucleus and cytoplasm of Xenopus oocytes. The data suggest new CRM1 functions in spatial control of vesicle coat-assembly, centrosomes, autophagy, peroxisome biogenesis, cytoskeleton, ribosome maturation, translation, mRNA degradation, and more generally in precluding a potentially detrimental action of cytoplasmic pathways within the nuclear interior. There are also numerous new instances where CRM1 appears to act in regulatory circuits. Altogether, our dataset allows unprecedented insights into the nucleocytoplasmic organisation of eukaryotic cells, into the contributions of an exceedingly promiscuous exportin and it provides a new basis for NES prediction.
- Published
- 2015
- Full Text
- View/download PDF
47. Ecto-5'-Nucleotidase Overexpression Reduces Tumor Growth in a Xenograph Medulloblastoma Model.
- Author
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Angélica R Cappellari, Micheli M Pillat, Hellio D N Souza, Fabrícia Dietrich, Francine H Oliveira, Fabrício Figueiró, Ana L Abujamra, Rafael Roesler, Joanna Lecka, Jean Sévigny, Ana Maria O Battastini, and Henning Ulrich
- Subjects
Medicine ,Science - Abstract
Ecto-5'-nucleotidase/CD73 (ecto-5'-NT) participates in extracellular ATP catabolism by converting adenosine monophosphate (AMP) into adenosine. This enzyme affects the progression and invasiveness of different tumors. Furthermore, the expression of ecto-5'-NT has also been suggested as a favorable prognostic marker, attributing to this enzyme contradictory functions in cancer. Medulloblastoma (MB) is the most common brain tumor of the cerebellum and affects mainly children.The effects of ecto-5'-NT overexpression on human MB tumor growth were studied in an in vivo model. Balb/c immunodeficient (nude) 6 to 14-week-old mice were used for dorsal subcutaneous xenograph tumor implant. Tumor development was evaluated by pathophysiological analysis. In addition, the expression patterns of adenosine receptors were verified.The human MB cell line D283, transfected with ecto-5'-NT (D283hCD73), revealed reduced tumor growth compared to the original cell line transfected with an empty vector. D283hCD73 generated tumors with a reduced proliferative index, lower vascularization, the presence of differentiated cells and increased active caspase-3 expression. Prominent A1 adenosine receptor expression rates were detected in MB cells overexpressing ecto-5'-NT.This work suggests that ecto-5'-NT promotes reduced tumor growth to reduce cell proliferation and vascularization, promote higher differentiation rates and initiate apoptosis, supposedly by accumulating adenosine, which then acts through A1 adenosine receptors. Therefore, ecto-5'-NT might be considered an important prognostic marker, being associated with good prognosis and used as a potential target for therapy.
- Published
- 2015
- Full Text
- View/download PDF
48. Hey bHLH Proteins Interact with a FBXO45 Containing SCF Ubiquitin Ligase Complex and Induce Its Translocation into the Nucleus.
- Author
-
Daniela Salat, Anja Winkler, Henning Urlaub, and Manfred Gessler
- Subjects
Medicine ,Science - Abstract
The Hey protein family, comprising Hey1, Hey2 and HeyL in mammals, conveys Notch signals in many cell types. The helix-loop-helix (HLH) domain as well as the Orange domain, mediate homo- and heterodimerization of these transcription factors. Although distinct interaction partners have been identified so far, their physiological relevance for Hey functions is still largely unclear. Using a tandem affinity purification approach and mass spectrometry analysis we identified members of an ubiquitin E3-ligase complex consisting of FBXO45, PAM and SKP1 as novel Hey1 associated proteins. There is a direct interaction between Hey1 and FBXO45, whereas FBXO45 is needed to mediate indirect Hey1 binding to SKP1. Expression of Hey1 induces translocation of FBXO45 and PAM into the nucleus. Hey1 is a short-lived protein that is degraded by the proteasome, but there is no evidence for FBXO45-dependent ubiquitination of Hey1. On the contrary, Hey1 mediated nuclear translocation of FBXO45 and its associated ubiquitin ligase complex may extend its spectrum to additional nuclear targets triggering their ubiquitination. This suggests a novel mechanism of action for Hey bHLH factors.
- Published
- 2015
- Full Text
- View/download PDF
49. Modulation of mouse embryonic stem cell proliferation and neural differentiation by the P2X7 receptor.
- Author
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Talita Glaser, Sophia La Banca de Oliveira, Arquimedes Cheffer, Renata Beco, Patrícia Martins, Maynara Fornazari, Claudiana Lameu, Helio Miranda Costa Junior, Robson Coutinho-Silva, and Henning Ulrich
- Subjects
Medicine ,Science - Abstract
BackgroundNovel developmental functions have been attributed to the P2X7 receptor (P2X7R) including proliferation stimulation and neural differentiation. Mouse embryonic stem cells (ESC), induced with retinoic acid to neural differentiation, closely assemble processes occurring during neuroectodermal development of the early embryo.Principal findingsP2X7R expression together with the pluripotency marker Oct-4 was highest in undifferentiated ESC. In undifferentiated cells, the P2X7R agonist Bz-ATP accelerated cell cycle entry, which was blocked by the specific P2X7R inhibitor KN-62. ESC induced to neural differentiation with retinoic acid, reduced Oct-4 and P2X7R expression. P2X7R receptor-promoted intracellular calcium fluxes were obtained at lower Bz-ATP ligand concentrations in undifferentiated and in neural-differentiated cells compared to other studies. The presence of KN-62 led to increased number of cells expressing SSEA-1, Dcx and β3-tubulin, as well as the number of SSEA-1 and β3-tubulin-double-positive cells confirming that onset of neuroectodermal differentiation and neuronal fate determination depends on suppression of P2X7R activity. Moreover, an increase in the number of Ki-67 positive cells in conditions of P2X7R inhibition indicates rescue of progenitors into the cell cycle, augmenting the number of neuroblasts and consequently neurogenesis.ConclusionsIn embryonic cells, P2X7R expression and activity is upregulated, maintaining proliferation, while upon induction to neural differentiation P2X7 receptor expression and activity needs to be suppressed.
- Published
- 2014
- Full Text
- View/download PDF
50. Rescue of amyloid-Beta-induced inhibition of nicotinic acetylcholine receptors by a peptide homologous to the nicotine binding domain of the alpha 7 subtype.
- Author
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Arthur A Nery, Margaret H Magdesian, Cleber A Trujillo, Luciana B Sathler, Maria A Juliano, Luiz Juliano, Henning Ulrich, and Sergio T Ferreira
- Subjects
Medicine ,Science - Abstract
Alzheimer's disease (AD) is characterized by brain accumulation of the neurotoxic amyloid-β peptide (Aβ) and by loss of cholinergic neurons and nicotinic acetylcholine receptors (nAChRs). Recent evidence indicates that memory loss and cognitive decline in AD correlate better with the amount of soluble Aβ than with the extent of amyloid plaque deposits in affected brains. Inhibition of nAChRs by soluble Aβ40 is suggested to contribute to early cholinergic dysfunction in AD. Using phage display screening, we have previously identified a heptapeptide, termed IQ, homologous to most nAChR subtypes, binding with nanomolar affinity to soluble Aβ40 and blocking Aβ-induced inhibition of carbamylcholine-induced currents in PC12 cells expressing α7 nAChRs. Using alanine scanning mutagenesis and whole-cell current recording, we have now defined the amino acids in IQ essential for reversal of Aβ40 inhibition of carbamylcholine-induced responses in PC12 cells, mediated by α7 subtypes and other endogenously expressed nAChRs. We further investigated the effects of soluble Aβ, IQ and analogues of IQ on α3β4 nAChRs recombinantly expressed in HEK293 cells. Results show that nanomolar concentrations of soluble Aβ40 potently inhibit the function of α3β4 nAChRs, and that subsequent addition of IQ or its analogues does not reverse this effect. However, co-application of IQ makes the inhibition of α3β4 nAChRs by Aβ40 reversible. These findings indicate that Aβ40 inhibits different subtypes of nAChRs by interacting with specific receptor domains homologous to the IQ peptide, suggesting that IQ may be a lead for novel drugs to block the inhibition of cholinergic function in AD.
- Published
- 2013
- Full Text
- View/download PDF
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