A5064035350- Cellular transport and secretion
- Calcium signaling and nucleotide metabolism
- COVID-19 Clinical Research Studies
- SARS-CoV-2 and COVID-19 Research
- Autophagy in Disease and Therapy
- Long-Term Effects of COVID-19
- PI3K/AKT/mTOR signaling in cancer
- Microtubule and mitosis dynamics
- SARS-CoV-2 detection and testing
- Lipid metabolism and biosynthesis
- Renal Diseases and Glomerulopathies
- Intensive Care Unit Cognitive Disorders
- Cellular Mechanics and Interactions
- Biosensors and Analytical Detection
- Lysosomal Storage Disorders Research
- Advanced Proteomics Techniques and Applications
- Glycosylation and Glycoproteins Research
- Lipid Membrane Structure and Behavior
- Pancreatic function and diabetes
- Biomedical Research and Pathophysiology
- Genetic and Kidney Cyst Diseases
- Anodic Oxide Films and Nanostructures
- Photosynthetic Processes and Mechanisms
- Heparin-Induced Thrombocytopenia and Thrombosis
- Endoplasmic Reticulum Stress and Disease
Innsbruck Medical University
2015-2025
Universität Innsbruck
2014-2024
University Hospital Innsbruck
2020
Co-operative College
2014
Vienna Biocenter
2013
Max Planck Unit for Structural Molecular Biology
2004
Ras GTPase-activating protein-binding proteins 1 and 2 (G3BP1 G3BP2, respectively) are widely recognized as core components of stress granules (SGs). We report that G3BPs reside at the cytoplasmic surface lysosomes. They act in a non-redundant manner to anchor tuberous sclerosis complex (TSC) protein lysosomes suppress activation metabolic master regulator mechanistic target rapamycin (mTORC1) by amino acids insulin. Like TSC complex, G3BP1 deficiency elicits phenotypes related mTORC1...
Abstract The transcription factor TFEB is a master regulator of lysosomal biogenesis and autophagy 1 . phosphorylation by the mechanistic target rapamycin complex (mTORC1) 2–5 unique in its mTORC1 substrate recruitment mechanism, which strictly dependent on amino acid-mediated activation RagC GTPase activating protein FLCN 6,7 lacks TOR signalling motif responsible for other substrates. We used cryogenic-electron microscopy to determine structure as presented phosphorylation, we refer...
Structure of human mTORC1 components The (mechanistic target rapamycin complex 1) garners much attention as a signaling hub that coordinates input from growth-factor receptors and nutrient availability with metabolism cell growth proliferation. de Araujo et al. report the crystal structure LAMTOR (or “Ragulator”) helps assemble at lysosomal membrane for activation. functional studies reveal how LAMTOR1 wraps around other subunits to hold them in place interacts Rag guanosine triphosphatases...
Signaling from lysosomes controls cellular clearance and energy metabolism. Lysosomal malfunction has been implicated in several pathologies, including neurodegeneration, cancer, infection, immunodeficiency, obesity. Interestingly, many functions are dependent on the organelle position. motility requires integration of extracellular intracellular signals that converge a competition between motor proteins ultimately control lysosomal movement microtubules. Here, we identify novel upstream...
Cell migration is mediated by the dynamic remodeling of focal adhesions (FAs). Recently, an important role endosomal signaling in regulation cell was recognized. Here, we show essential function for late endosomes carrying p14-MP1 (LAMTOR2/3) complex FA dynamics. p14-MP1-positive move to periphery along microtubules (MTs) a kinesin1- and Arl8b-dependent manner. There they specifically target FAs regulate turnover, which required migration. Using genetically modified fibroblasts from...
Lp(a) concentrations represent a major cardiovascular risk factor and are almost entirely controlled by one single locus (LPA). However, many genetic factors in LPA governing the enormous variance of levels still unknown. Since up to 70% coding sequence located difficult access hypervariable copy number variation named KIV-2, we hypothesized that it may contain novel functional variants with pronounced effects on concentrations. We performed large scale mutation analysis KIV-2 using an...
Signaling pathways in eukaryotic cells are often controlled by the formation of specific signaling complexes, which coordinated scaffold and adaptor proteins. Elucidating their molecular architecture is essential to understand spatial temporal regulation cellular signaling. p14 MP1 form a tight ( K d = 12.8 nM) endosomal adaptor/scaffold complex, regulates mitogen-activated protein kinase (MAPK) Here, we present 1.9-Å crystal structure biologically functional p14/MP1 complex. The overall...
The late endosomal adaptor protein LAMTOR2/p14 is essential for tissue homeostasis by controlling MAPK and mTOR signaling, which in turn regulate cell growth proliferation, migration spreading. Moreover, LAMTOR2 critically controls architecture function of the endocytic system, including epidermal factor receptor (EGFR) degradation lysosomes, positioning endosomes defense against intracellular pathogens. Here we describe multifaceted ultrastructural phenotype endo/lysosomal system...
Density gradient centrifugation is a common method for separating intracellular organelles. During centrifugation, organelles float or sediment until they reach their isopycnic position within the gradient. The density of an organelle depends on its content, size, shape, and lipid:protein ratio. degree separation between different will therefore be highly dependent how points are in given buffer. Separation also depend medium used to prepare gradient, whether it sucrose (the most common)...
Abstract Heterozygous mutations in the gene encoding RagD GTPase were shown to cause a novel autosomal dominant condition characterized by kidney tubulopathy and cardiomyopathy. We previously demonstrated that RagD, its paralogue RagC, mediate non-canonical mTORC1 signaling pathway inhibits activity of TFEB TFE3, transcription factors MiT/TFE family master regulators lysosomal biogenesis autophagy. Here we show causing cardiomyopathy are “auto- activating”, even absence Folliculin, GAP...
ABSTRACT Cells experience oxidative stress and widespread cellular damage during stress‐induced premature senescence (SIPS). Senescent cells show an increase in lysosomal content, which may contribute to mitigating by promoting autophagy. This study investigates the dynamics of quality control human dermal fibroblasts (HDF), specifically examining signaling pathways SIPS. Our results reveal distinct responses between initial phase ensuing senescent phenotype. During phase, treatment with...
The neurodevelopmental disorders Prader-Willi syndrome (PWS) and Schaaf-Yang (SYS) both arise from genomic alterations within human chromosome 15q11–q13. A deletion of the SNORD116 cluster, encoding small nucleolar RNAs, or frameshift mutations MAGEL2 result in closely related phenotypes individuals with PWS SYS, respectively. By investigation their subcellular localization, we observed that contrast to a predominant cytoplasmic localization wild-type (WT) MAGEL2, truncated mutant was evenly...
Homogenization is the name given to methodological steps necessary for releasing organelles and other cellular constituents as a free suspension of intact individual components. Most homogenization procedures used mammalian cells (e.g., cavitation pump Dounce homogenizer) rely on mechanical force break plasma membrane may be supplemented with osmotic or temperature alterations facilitate disruption. In this protocol, we describe syringe-based method that does not require specialized...
Abstract The diagnosis of COVID-19 relies on the direct detection SARS-CoV-2 RNA in respiratory specimens by RT-PCR. pandemic spread disease caused an imbalance between demand and supply materials reagents needed for diagnostic purposes including swab sets. In a comparative effectiveness study, we conducted serial follow-up swabs hospitalized laboratory-confirmed patients. We assessed performance in-house system developed according to recommendations US CDC. total 96 swabs, found significant...
Immunogold labeling of permeabilized whole‐mount cells or thin‐sectioned material is widely used for the subcellular localization biomolecules at high spatial resolution electron microscopy (EM). Those approaches are well compatible with either 3‐dimensional (3D) reconstruction organelle morphology and antigen distribution rapid cryofixation—but not easily both once. We describe here a specimen preparation protocol animal cell cultures, which represents novel blend specifically adapted...
The successful combination of highly sensitive mass spectrometry and pre-fractionation techniques has provided a powerful tool to detect dynamic changes in low abundant regulatory proteins at the organelle level. Subcellular fractionation, being flexible, adjustable (both cell tissues), allowing analysis their physiologic/ intracellular context, become most commonly used preparative/enrichment method. This chapter introduces state-of-the-art subcellular fractionation protocols briefly...
LAMTOR2 (p14), a part of the larger LAMTOR/Ragulator complex, plays crucial role in EGF-dependent activation p42/44 mitogen-activated protein kinases (MAPK, ERK1/2). In this study, we investigated nerve growth factor (NGF)-mediated neuronal differentiation. Stimulation PC12 (rat adrenal pheochromocytoma) cells with NGF is known to activate MAPK. Pharmacological inhibition MEK1 as well siRNA–mediated knockdown both p42 and p44 MAPK resulted neurite outgrowth. Contrary expectations,...
Immortalized macrophage lines and primary macrophages display the ability to internalize small latex beads through endocytic pathway. This protocol describes a simple robust method for separating organelles from on sucrose gradient, taking advantage of significantly lower density containing compared with other intracellular organelles. The are retained in endosomes as they mature; therefore, harvesting cells at different time points after internalization permits purification organelle...
Seizure threshold 2 (SZT2) is a component of the KICSTOR complex which, under catabolic conditions, functions as negative regulator in amino acid-sensing branch mTORC1. Mutations this gene cause severe neurodevelopmental and epileptic encephalopathy whose main symptoms include epilepsy, intellectual disability, macrocephaly. As SZT2 remains one least characterized regulators mTORC1, work we performed systematic interactome analysis anabolic conditions. Besides numerous mTORC1 AMPK signaling...
Proteomic analysis of early and late endosomes has been constrained by the limited purity endosomal fractions that can be achieved biochemical methods. Here we briefly review endocytic pathways, then introduce fractionation strategies have used to improve isolated endosomes. In addition, describe innovative proteomics methods shown partially circumvent limitations found in enrichment steps.