10 results on '"Jucht, Agnieszka E."'
Search Results
2. Oxomer- and Reporter Gene-Based Analysis of FIH Activity in Cells
- Author
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Volkova, Yulia L., primary, Jucht, Agnieszka E., additional, and Scholz, Carsten C., additional
- Published
- 2024
- Full Text
- View/download PDF
3. Oxomer- and Reporter Gene-Based Analysis of FIH Activity in Cells
- Author
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Gilkes, Daniele M, Gilkes, D M ( Daniele M ), Volkova, Yulia L; https://orcid.org/0000-0001-5772-2889, Jucht, Agnieszka E; https://orcid.org/0000-0002-4553-4319, Scholz, Carsten C; https://orcid.org/0000-0001-6579-8015, Gilkes, Daniele M, Gilkes, D M ( Daniele M ), Volkova, Yulia L; https://orcid.org/0000-0001-5772-2889, Jucht, Agnieszka E; https://orcid.org/0000-0002-4553-4319, and Scholz, Carsten C; https://orcid.org/0000-0001-6579-8015
- Published
- 2024
4. Selective Hypoxia-Sensitive Oxomer Formation by FIH Prevents Binding of the NF-κB Inhibitor IκBβ to NF-κB Subunits.
- Author
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Volkova, Yulia L., Jucht, Agnieszka E., Oechsler, Nina, Krishnankutty, Roopesh, von Kriegsheim, Alex, Wenger, Roland H., and Scholz, Carsten C.
- Subjects
- *
OXYGEN detectors , *AMINO acid sequence - Abstract
Pharmacologic inhibitors of cellular hydroxylase oxygen sensors are protective in multiple preclinical in vivo models of inflammation. However, the molecular mechanisms underlying this regulation are only partly understood, preventing clinical translation. We previously proposed a new mechanism for cellular oxygen sensing: oxygen-dependent, (likely) covalent protein oligomer (oxomer) formation. Here, we report that the oxygen sensor factor inhibiting HIF (FIH) forms an oxomer with the NF-κB inhibitor β (IκBβ). The formation of this protein complex required FIH enzymatic activity and was prevented by pharmacologic inhibitors. Oxomer formation was highly hypoxia-sensitive and very stable. No other member of the IκB protein family formed an oxomer with FIH, demonstrating that FIH-IκBβ oxomer formation was highly selective. In contrast to the known FIH-dependent oxomer formation with the deubiquitinase OTUB1, FIH-IκBβ oxomer formation did not occur via an IκBβ asparagine residue, but depended on the amino acid sequence VAERR contained within a loop between IκBβ ankyrin repeat domains 2 and 3. Oxomer formation prevented IκBβ from binding to its primary interaction partners p65 and c-Rel, subunits of NF-κB, the master regulator of the cellular transcriptional response to pro-inflammatory stimuli. We therefore propose that FIH-mediated oxomer formation with IκBβ contributes to the hypoxia-dependent regulation of inflammation. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
5. The Asparagine Hydroxylase FIH: A Unique Oxygen Sensor
- Author
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Volkova, Yulia L., primary, Pickel, Christina, additional, Jucht, Agnieszka E., additional, Wenger, Roland H., additional, and Scholz, Carsten C., additional
- Published
- 2022
- Full Text
- View/download PDF
6. The Deubiquitinase OTUB1 Is a Key Regulator of Energy Metabolism
- Author
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Ruiz-Serrano, Amalia, Boyle, Christina N; https://orcid.org/0000-0002-1060-2529, Monné Rodríguez, Josep M; https://orcid.org/0000-0002-2574-0780, Günter, Julia; https://orcid.org/0000-0002-9397-5653, Jucht, Agnieszka E; https://orcid.org/0000-0002-4553-4319, Pfundstein, Svende, Bapst, Andreas M, Lutz, Thomas A; https://orcid.org/0000-0002-5056-8548, Wenger, Roland H; https://orcid.org/0000-0001-7592-4839, Scholz, Carsten C; https://orcid.org/0000-0001-6579-8015, Ruiz-Serrano, Amalia, Boyle, Christina N; https://orcid.org/0000-0002-1060-2529, Monné Rodríguez, Josep M; https://orcid.org/0000-0002-2574-0780, Günter, Julia; https://orcid.org/0000-0002-9397-5653, Jucht, Agnieszka E; https://orcid.org/0000-0002-4553-4319, Pfundstein, Svende, Bapst, Andreas M, Lutz, Thomas A; https://orcid.org/0000-0002-5056-8548, Wenger, Roland H; https://orcid.org/0000-0001-7592-4839, and Scholz, Carsten C; https://orcid.org/0000-0001-6579-8015
- Abstract
Dysregulated energy metabolism is a major contributor to a multitude of pathologies, including obesity and diabetes. Understanding the regulation of metabolic homeostasis is of utmost importance for the identification of therapeutic targets for the treatment of metabolically driven diseases. We previously identified the deubiquitinase OTUB1 as substrate for the cellular oxygen sensor factor-inhibiting HIF (FIH) with regulatory effects on cellular energy metabolism, but the physiological relevance of OTUB1 is unclear. Here, we report that the induced global deletion of OTUB1 in adult mice (Otub1 iKO) elevated energy expenditure, reduced age-dependent body weight gain, facilitated blood glucose clearance and lowered basal plasma insulin levels. The respiratory exchange ratio was maintained, indicating an unaltered nutrient oxidation. In addition, Otub1 deletion in cells enhanced AKT activity, leading to a larger cell size, higher ATP levels and reduced AMPK phosphorylation. AKT is an integral part of insulin-mediated signaling and Otub1 iKO mice presented with increased AKT phosphorylation following acute insulin administration combined with insulin hypersensitivity. We conclude that OTUB1 is an important regulator of metabolic homeostasis.
- Published
- 2022
7. OTUB1 regulates lung development, adult lung tissue homeostasis, and respiratory control
- Author
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Ruiz‐Serrano, Amalia, primary, Monné Rodríguez, Josep M., additional, Günter, Julia, additional, Sherman, Samantha P. M., additional, Jucht, Agnieszka E., additional, Fluechter, Pascal, additional, Volkova, Yulia L., additional, Pfundstein, Svende, additional, Pellegrini, Giovanni, additional, Wagner, Carsten A., additional, Schneider, Christoph, additional, Wenger, Roland H., additional, and Scholz, Carsten C., additional
- Published
- 2021
- Full Text
- View/download PDF
8. OTUB1 regulates lung development, adult lung tissue homeostasis, and respiratory control
- Author
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Ruiz-Serrano, Amalia, Monné Rodríguez, Josep M, Günter, Julia, Sherman, Samantha P M, Jucht, Agnieszka E; https://orcid.org/0000-0002-4553-4319, Fluechter, Pascal, Volkova, Yulia L; https://orcid.org/0000-0001-5772-2889, Pfundstein, Svende, Pellegrini, Giovanni, Wagner, Carsten A; https://orcid.org/0000-0002-9874-8898, Schneider, Christoph, Wenger, Roland H; https://orcid.org/0000-0001-7592-4839, Scholz, Carsten C; https://orcid.org/0000-0001-6579-8015, Ruiz-Serrano, Amalia, Monné Rodríguez, Josep M, Günter, Julia, Sherman, Samantha P M, Jucht, Agnieszka E; https://orcid.org/0000-0002-4553-4319, Fluechter, Pascal, Volkova, Yulia L; https://orcid.org/0000-0001-5772-2889, Pfundstein, Svende, Pellegrini, Giovanni, Wagner, Carsten A; https://orcid.org/0000-0002-9874-8898, Schneider, Christoph, Wenger, Roland H; https://orcid.org/0000-0001-7592-4839, and Scholz, Carsten C; https://orcid.org/0000-0001-6579-8015
- Abstract
OTUB1 is one of the most highly expressed deubiquitinases, counter-regulating the two most abundant ubiquitin chain types. OTUB1 expression is linked to the development and progression of lung cancer and idiopathic pulmonary fibrosis in humans. However, the physiological function of OTUB1 is unknown. Here, we show that constitutive whole-body Otub1 deletion in mice leads to perinatal lethality by asphyxiation. Analysis of (single-cell) RNA sequencing and proteome data demonstrated that OTUB1 is expressed in all lung cell types with a particularly high expression during late-stage lung development (E16.5, E18.5). At E18.5, the lungs of animals with Otub1 deletion presented with increased cell proliferation that decreased saccular air space and prevented inhalation. Flow cytometry-based analysis of E18.5 lung tissue revealed that Otub1 deletion increased proliferation of major lung parenchymal and mesenchymal/other non-hematopoietic cell types. Adult mice with conditional whole-body Otub1 deletion (wbOtub1del/del ) also displayed increased lung cell proliferation in addition to hyperventilation and failure to adapt the respiratory pattern to hypoxia. On the molecular level, Otub1 deletion enhanced mTOR signaling in embryonic and adult lung tissues. Based on these results, we propose that OTUB1 is a negative regulator of mTOR signaling with essential functions for lung cell proliferation, lung development, adult lung tissue homeostasis, and respiratory regulation. Keywords: FIH; HIF1AN; deubiquitinating enzyme; hypoxia; respiratory distress syndrome; respiratory failure.
- Published
- 2021
9. Oxomer- and Reporter Gene-Based Analysis of FIH Activity in Cells.
- Author
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Volkova YL, Jucht AE, and Scholz CC
- Subjects
- Humans, Genes, Reporter, Mixed Function Oxygenases, Hypoxia, Oxygen, Deubiquitinating Enzymes, Erythropoietin genetics
- Abstract
Cellular and tissue adaptations to oxygen deprivation (hypoxia) are necessary for both normal physiology and disease. Responses to hypoxia are initiated by the cellular oxygen sensors prolyl-4-hydroxylase domain (PHD) proteins 1-3 and factor inhibiting HIF (FIH). These enzymes regulate the transcription factor hypoxia-inducible factor (HIF) in a hypoxia-sensitive manner. FIH also regulates proteins outside the HIF pathway, including the deubiquitinase OTUB1. Numerous preclinical analyses have demonstrated that treatment with HIF hydroxylase inhibitors is beneficial and protective in many hypoxia-associated diseases. However, clinically available HIF hydroxylase inhibitors increase erythropoietin (EPO) gene expression and red blood cell production, which can be detrimental in hypoxia-associated conditions, such as ischemia/reperfusion injury of the heart or chronic inflammation. Our understanding of the relevance of FIH in (patho)physiology is only in its infancy, but FIH activity does not govern erythropoietin expression. Therefore, it is of prime interest to assess the relevance of FIH activity in (patho)physiology in detail, as it may contribute to developing novel therapeutic options for treating hypoxia-associated diseases that do not affect Epo regulation. Here, we describe specific protocols for two different methods to assess FIH enzymatic activity within cells, using a HIF-dependent firefly luciferase-reporter gene and an oxomer-dependent assay. Oxomers are oxygen-dependent stable protein oligomers formed by FIH, for example, with the deubiquitinase OTUB1. Oxomer formation directly depends on FIH activity, providing a suitable cellular readout for an easy-to-use analysis of FIH enzymatic activity in cellulo. These techniques permit an analysis of FIH activity toward HIF and outside the HIF pathway, allowing the investigation of FIH activity under different (patho)physiological conditions and assessment of novel (putative) inhibitors., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
- Published
- 2024
- Full Text
- View/download PDF
10. The Deubiquitinase OTUB1 Is a Key Regulator of Energy Metabolism.
- Author
-
Ruiz-Serrano A, Boyle CN, Monné Rodríguez JM, Günter J, Jucht AE, Pfundstein S, Bapst AM, Lutz TA, Wenger RH, and Scholz CC
- Subjects
- Adenylate Kinase metabolism, Animals, Blood Glucose, Body Weight, Cell Size, Cells, Cultured, Cysteine Endopeptidases metabolism, Energy Metabolism, Fibroblasts cytology, Fibroblasts drug effects, Fibroblasts metabolism, Insulin adverse effects, Mice, Phosphorylation, Proto-Oncogene Proteins c-akt metabolism, Adenosine Triphosphate metabolism, Cysteine Endopeptidases genetics, Gene Deletion, Insulin administration & dosage, Insulin Resistance genetics, Mixed Function Oxygenases metabolism
- Abstract
Dysregulated energy metabolism is a major contributor to a multitude of pathologies, including obesity and diabetes. Understanding the regulation of metabolic homeostasis is of utmost importance for the identification of therapeutic targets for the treatment of metabolically driven diseases. We previously identified the deubiquitinase OTUB1 as substrate for the cellular oxygen sensor factor-inhibiting HIF (FIH) with regulatory effects on cellular energy metabolism, but the physiological relevance of OTUB1 is unclear. Here, we report that the induced global deletion of OTUB1 in adult mice ( Otub1 iKO) elevated energy expenditure, reduced age-dependent body weight gain, facilitated blood glucose clearance and lowered basal plasma insulin levels. The respiratory exchange ratio was maintained, indicating an unaltered nutrient oxidation. In addition, Otub1 deletion in cells enhanced AKT activity, leading to a larger cell size, higher ATP levels and reduced AMPK phosphorylation. AKT is an integral part of insulin-mediated signaling and Otub1 iKO mice presented with increased AKT phosphorylation following acute insulin administration combined with insulin hypersensitivity. We conclude that OTUB1 is an important regulator of metabolic homeostasis.
- Published
- 2022
- Full Text
- View/download PDF
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