DOCSLIB.ORG

Loss of a Subunit of Vacuolar Atpase Identifies Unexpected Biological Signatures of Reduced Organelle Acidification in Vivo

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Loss of a Subunit of Vacuolar Atpase Identifies Unexpected Biological Signatures of Reduced Organelle Acidification in Vivo bioRxiv preprint doi: https://doi.org/10.1101/2020.11.30.397463; this version posted November 30, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Loss of a subunit of vacuolar ATPase identifies unexpected biological signatures of reduced organelle acidification in vivo Short title: Loss of vacuolar ATPase function and reduced acidification in vivo Pottie Lore1,2, Van Gool Wouter1,2, Vanhooydonck Michiel1,2, Hanisch Franz-Georg3, Goeminne Geert4,5, Rajkovic Andreja 6, Coucke Paul1,2, Sips Patrick1,2* and Callewaert Bert1,2*. 1. Center for Medical Genetics Ghent, Ghent University Hospital, 9000 Ghent, Belgium 2. Department of Biomolecular Medicine, Ghent University, 9000 Ghent 3. Institute of Biochemistry II, Medical Faculty, University of Cologne, 50931 Köln, Germany. 4. VIB Metabolomics Core Ghent, 9052 Ghent, Belgium. 5. Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium. 6. Department of Food technology, Safety and Health, Faculty of Bioscience Engineering, University of Ghent, 9000 Ghent, Belgium * Sips P and Callewaert B contributed equally to this work. Corresponding author Callewaert Bert Center for Medical Genetics Ghent University Hospital Corneel Heymanslaan 10 9000 Gent, Belgium [email protected] Keywords ATP6V1E1, danio rerio, acidification, transcriptome, metabolome, sphingolipids, endo(lyso)somes Funding B.C. is a senior clinical investigator of the Research Foundation Flanders. This work was supported by a starting grant of the Special Research Fund, Flanders of Ghent University (grant 01N04516C to B.C.), by a Methusalem Grant (BOFMET2015000401) from Ghent University, by a junior fundamental research project grant of the Fund for Scientific Research (G035620N) and by the European Union’s Horizon 2020 research and innovation program, under the Marie Skłodowska-Curie grant agreement No. 794365 to P.S. Conflicts of Interest: The authors declare no conflict of interest bioRxiv preprint doi: https://doi.org/10.1101/2020.11.30.397463; this version posted November 30, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract The inability to maintain a strictly regulated endo(lyso)somal acidic pH through the proton-pumping action of the vacuolar-ATPases has been associated with various human diseases including heritable connective tissue disorders, neurodegenerative diseases and cancer. Multiple studies have investigated the pleiotropic effects of reduced acidification in vitro, but the mechanisms elicited by impaired endo(lyso)somal acidification in vivo remain poorly understood. Here, we show that loss of atp6v1e1b in zebrafish leads to early mortality, associated with craniofacial dysmorphisms, vascular anomalies, cardiac dysfunction, hypotonia and epidermal structural defects, reminiscent of the phenotypic manifestations in cutis laxa patients carrying a defect in the ATP6V1E1 gene. Mechanistically, we found that in vivo genetic depletion of atp6v1e1b leads to N-glycosylation defects and reduced maturation of endosomal and lysosomal vesicles, but retains the hypoxia-mediated response. In order to gain further insights into the processes affected by aberrant organelle acidification, we performed an untargeted analysis of the transcriptome and metabolome in early atp6v1e1b-deficient larvae. We report multiple affected pathways including but not limited to oxidative phosphorylation, sphingolipid and fatty acid metabolism with profound defects in mitochondrial respiration. Taken together, our results identify new complex biological effects of reduced organelle acidification in vivo, which likely contribute to the multisystemic manifestations observed in disorders caused by v-ATPase deficiency. Introduction The intracellular pH of specific subcellular compartments is maintained by the proton-pumping action of the vacuolar-ATPases (v-ATPase), thereby regulating a range of molecular processes including activation of enzyme activity, protein folding, vesicle trafficking, and support of organelle function and integrity. V-ATPases consist of ubiquitously expressed multisubunit complexes, composed of a membrane-associated V0 domain and a cytosolic (catalytic) V1 domain. Hydrolysis of ATP by the complex generates proton pumping activity against a concentration gradient in the lumen of endo(lyso)somes, secretory vesicles, Golgi apparatus and across the plasma membrane of specialized cells including osteoclasts, epididymal clear cells and renal epithelial intercalating cells [1]. Various human diseases are linked to defects in genes encoding subunits of the v-ATPase, including neurological disorders (e.g. encephalopathy with epilepsy (ATP6V1A [MIM: 618012]), X-linked mental retardation Hedera type (ATP6AP2 [MIM: 300423]), and X-linked Parkinson Disease with spasticity (ATP6AP2 [MIM: 300911])), congenital disorders of glycosylation (ATP6AP1 [MIM: 300972] and ATP6AP2 [MIM: 301045]), disorders in which the secretion of protons in the extracellular environment is disrupted (e.g. distal renal tubular acidosis and hearing loss (ATP6V1B1 [MIM: 192132] and ATP6V0A4 [MIM: 605239]), syndromic osteopetrosis (ATP6V0A3 [MIM:259700]), Zimmermann- Laband syndrome (ATP6V1B2 [MIM: 616455]), dominant deafness-onychodystrophy syndrome (ATP6V1B2 [MIM: 124480])) and connective tissue disorders (e.g. autosomal recessive (AR) cutis laxa (CL) syndrome type 2A (ATP6V0A2 [MIM: 219200]), ARCL type 2C (ATP6V1E1 [MIM: 617402]), and ARCL type 2D (ATP6V1A [MIM: 617403])) [2-12]. ARCL type 2 shows variable glycosylation bioRxiv preprint doi: https://doi.org/10.1101/2020.11.30.397463; this version posted November 30, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. abnormalities [10, 13, 14], and is characterized by skin wrinkles or loose redundant skin folds, variable level of mental impairment, delayed neuromotor development, sensorineural hearing loss, skeletal abnormalities, facial dysmorphology, hypotonia, and cardiopulmonary involvement including pneumothorax, hypertrophic cardiomyopathy, and aortic root dilation [10, 11, 15, 16]. Finally, v- ATPases have been implicated in cancer growth and metastasis in a variety of contexts [17]. Higher v- ATPase expression and relocation to the plasma membrane is observed in proliferating cancer cells including breast, prostate, liver, pancreatic, melanoma, and esophageal cancers [18]. V-ATPase activity in the plasma membrane promotes cell survival since the resulting alkalization of the intracellular environment supports the Warburg effect by reducing the excess acid load caused by glycolytic lactic acid production [19]. Intracellular v-ATPases further regulate key physiological cell signaling pathways. V-ATPase are involved in downstream Notch signaling [20-23], Wnt signaling [18, 24-29], activation of mechanistic target of rapamycin complex 1 (mTORC1), and AMP-activated protein kinase (AMPK) signaling [17]. Recent studies have highlighted a novel mechanism linking v-ATPase inhibition, hypoxia-inducible factor (HIF) signaling and iron deficiency. Using in vitro experiments, it was shown that genetic and pharmacological inhibition of v-ATPases impairs endo(lyso)somal acidification which leads to defective cellular iron uptake via the transferrin system. Iron depletion results in reduced prolyl hydroxylase activity, leading to stabilization of HIF and activation of HIF-dependent signaling mechanisms [30-32]. This raises the question whether in vivo v-ATPase deficiency impairs iron homeostasis and perturbates HIF-mediated response, and if this represents the major mode of action in genetic disorders related to defects in v-ATPase subunits. Complete loss of the murine proteolipid 16 kDa subunit of the v-ATPase V0 complex is embryonically lethal [33]. Since external fertilization and transparent development allow us to study embryonic development in zebrafish, we modelled genetically depleted atp6v1e1b zebrafish. Despite early mortality, this model mimics ARCL type 2C with hypotonia, craniofacial abnormalities, vascular anomalies, and an altered dermal connective tissue structure. Untargeted analysis of the transcriptome, metabolome, and lipidome furthermore indicates alterations in oxidative phosphorylation, fatty acid metabolism, and sphingolipid metabolism in this in vivo model of v-ATPase deficiency. Results Genetic loss of atp6v1e1b in zebrafish leads to early mortality and mimics the human phenotype of ATP6V1E1-related cutis laxa syndrome. We genetically depleted atp6v1e1b, a subunit of the v-ATPase that connects the transmembrane domain with the cytosolic domain, in zebrafish to identify pleiotropic manifestations of reduced organelle acidification in vivo. We generated a zebrafish line harboring a two base-pair insertion followed by a three base-pair deletion in exon 5 of atp6v1e1b (atp6v1e1bcmg78/+), c.334insGG; c.337-340delCGG, which led to nonsense-mediated decay (NMD) due to a premature stop codon (Supplemental Figure 1A-B). We further obtained a zebrafish line with viral insertion in the 5’UTR of atp6v1e1b bioRxiv preprint doi: https://doi.org/10.1101/2020.11.30.397463; this version posted November 30, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder.
Load more

Recommended publications
  • Mt-Atp8 Gene in the Conplastic Mouse Strain C57BL/6J-Mtfvb/NJ on the Mitochondrial Function and Consequent Alterations to Metabolic and Immunological Phenotypes
    From the Lübeck Institute of Experimental Dermatology of the University of Lübeck Director: Prof. Dr. Saleh M. Ibrahim Interplay of mtDNA, metabolism and microbiota in the pathogenesis of AIBD Dissertation for Fulfillment of Requirements for the Doctoral Degree of the University of Lübeck from the Department of Natural Sciences Submitted by Paul Schilf from Rostock Lübeck, 2016 First referee: Prof. Dr. Saleh M. Ibrahim Second referee: Prof. Dr. Stephan Anemüller Chairman: Prof. Dr. Rainer Duden Date of oral examination: 30.03.2017 Approved for printing: Lübeck, 06.04.2017 Ich versichere, dass ich die Dissertation ohne fremde Hilfe angefertigt und keine anderen als die angegebenen Hilfsmittel verwendet habe. Weder vorher noch gleichzeitig habe ich andernorts einen Zulassungsantrag gestellt oder diese Dissertation vorgelegt. ABSTRACT Mitochondria are critical in the regulation of cellular metabolism and influence signaling processes and inflammatory responses. Mitochondrial DNA mutations and mitochondrial dysfunction are known to cause a wide range of pathological conditions and are associated with various immune diseases. The findings in this work describe the effect of a mutation in the mitochondrially encoded mt-Atp8 gene in the conplastic mouse strain C57BL/6J-mtFVB/NJ on the mitochondrial function and consequent alterations to metabolic and immunological phenotypes. This work provides insights into the mutation-induced cellular adaptations that influence the inflammatory milieu and shape pathological processes, in particular focusing on autoimmune bullous diseases, which have recently been reported to be associated with mtDNA polymorphisms in the human MT-ATP8 gene. The mt-Atp8 mutation diminishes the assembly of the ATP synthase complex into multimers and decreases mitochondrial respiration, affects generation of reactive oxygen species thus leading to a shift in the metabolic balance and reduction in the energy state of the cell as indicated by the ratio ATP to ADP.
    [Show full text]
  • Systemic Hypoxia Inhibits T Cell Response by Limiting Mitobiogenesis
    RESEARCH ARTICLE Systemic hypoxia inhibits T cell response by limiting mitobiogenesis via matrix substrate-level phosphorylation arrest Amijai Saragovi1, Ifat Abramovich2, Ibrahim Omar1, Eliran Arbib1, Ori Toker3, Eyal Gottlieb2, Michael Berger1* 1The Lautenberg center for Immunology and Cancer Research, The Institute for Medical Research Israel-Canada, The Hebrew University Medical School, Jerusalem, Israel; 2The Ruth and Bruce Rappaport, Faculty of Medicine, Technion - Israel Institute of Technology, Jerusalem, Israel; 3Faculty of Medicine, Hebrew University of Jerusalem; The Allergy and Immunology Unit, Shaare Zedek Medical Center, Jerusalem, Israel Abstract Systemic oxygen restriction (SOR) is prevalent in numerous clinical conditions, including chronic obstructive pulmonary disease (COPD), and is associated with increased susceptibility to viral infections. However, the influence of SOR on T cell immunity remains uncharacterized. Here we show the detrimental effect of hypoxia on mitochondrial-biogenesis in activated mouse CD8+ T cells. We find that low oxygen level diminishes CD8+ T cell anti-viral response in vivo. We reveal that respiratory restriction inhibits ATP-dependent matrix processes that are critical for mitochondrial-biogenesis. This respiratory restriction-mediated effect could be rescued by TCA cycle re-stimulation, which yielded increased mitochondrial matrix-localized ATP via substrate-level phosphorylation. Finally, we demonstrate that the hypoxia-arrested CD8+ T cell anti-viral response could be rescued in vivo through brief exposure to atmospheric oxygen pressure. Overall, these findings elucidate the detrimental effect of hypoxia on mitochondrial- + *For correspondence: biogenesis in activated CD8 T cells, and suggest a new approach for reducing viral infections in [email protected] COPD. Competing interests: The authors declare that no competing interests exist.
    [Show full text]
  • Effect of Phosphate and Uncouplers on Substrate Transport And
    Plant Physiol. (1977) 59, 139-144 Effect of Phosphate and Uncouplers on Substrate Transport and Oxidation by Isolated Corn Mitochondria1 Received for publication May 25, 1976 and in revised form September 13, 1976 DAVID A. DAY2 AND JOHN B. HANSON Department of Botany, University of Illinois, Urbana, Illinois 61801 ABSTRACT ADP) stimulates the rate by increasing transport (3, 25, 27). However, Pi stimulation of exogenous NADH oxidation by corn A study was made to determine conditions under which malate oxida- mitochondria has also been reported (9). Oxidation of this tion rates in corn (Zea mays L.) mitochondria are limited by transport substrate does not require its penetration of the inner membrane processes. In the absence of added ADP, inorganic phosphate increased (3, 6) and the Pi stimulation was attributed to an accelerated malate oxidation rates by processes inhibited by mersalyl and oligomy- turnover of the coupling mechanism, since it was sensitive to cin, but phosphate did not stimulate uncoupled respiration. However, oligomycin (9). Oligomycin did not inhibit Pi stimulation of the uncoupled oxidation rates were inhibited by butylmalonate and malate oxidation by cauliflower mitochondria (27). mersalyl. When uncoupler was added prior to substrate, subsequent 02 Inhibition of substrate transport by uncouplers has been ob- uptake rates were reduced when malate and succinate, but not exoge- served with animal mitochondria (23), but the low rates of nous NADH, were used. Uncoupler and butylmalonate also inhibited substrate oxidation by plant mitochondria in the presence of swelling in malate solutions and malate accumulation by these mitochon- uncouplers have generally been attributed to a requirement for dria, which were found to have a high endogenous phosphate content.
    [Show full text]
  • F1-Atpase-Catalyzed Synthesis of ATP from Oleoylphosphate and ADP (Mitochondria/Adenosine Triphosphate) RICHARD JOHNSTON and RICHARD S
    Proc. Natl. Acad. Sci. USA Vol. 74, No. 11, pp. 4919-4923, November 1977 Biochemistry F1-ATPase-catalyzed synthesis of ATP from oleoylphosphate and ADP (mitochondria/adenosine triphosphate) RICHARD JOHNSTON AND RICHARD S. CRIDDLE* Department of Biochemistry and Biophysics, University of California, Davis, California 95616 Communicated by Paul D. Boyer, August 29,1977 ABSTRACT Purified preparations of F1-ATPase (ATP The specific activities of purified preparations of oligomy- phos hohydrolase; EC 3.6.1.3) isolated from yeast mitochondria cin-sensitive ATPase were in the range of 15-20 ,qmole of ATP cata yze the reaction of oleoylphosphate with ADP to yield ATP and oleic acid. Formation of ATP is specifically inhibited by hydrolyzed/min per mg of protein. Specific activities of F1 the F1-ATPase inhibitor 1799 and by dinitrophenol. In the preparations were near 33 umol of ATP hydrolyzed/min per presence of Fi, dinitrophenol "uncouples" the synthase reaction mg. ATPase assays were performed by the coupled spectro- by causing rapid hydrolysis of oleoylphosphate without ATP photometric method of Monroy and Pullman (8) in pH 7.4 formation. It is propse that this Fl-catalyzed ATP synthesis Tris-HCI buffer containing 6 mM MgCI2. Oligomycin, dini- reaction corresponds to the terminal chemical step in oxidative trophenol, and 1799 were added as methanolic solutions. phosphorylation. Oleoylphosphate was prepared by both the methods de- D. Griffiths and coworkers have recently reported ATP syn- scribed by Lehninger (9) and by Hildebrand and Spector (10). thesis catalyzed by preparations of submitochondrial particles Alternatively, oleoylphosphate was prepared by a combination and by the oligomycin-sensitive ATPase from beef heart or of these two methods.
    [Show full text]
  • Characterizing a Better Uncoupler
    APPLICATION NOTE Characterizing a Better Uncoupler Cellular Metabolism Services Group Cayman Chemical Company Key Features · The novel compound BAM15 uncouples oxidative phosphorylation, depleting mitochondrial membrane potential at a wider effective concentration range than the classical uncoupler FCCP. · BAM15 and FCCP uncoupling properties are compared in HCT116, C2C12, and Huh7 cell lines using the Seahorse XF assay to determine maximal oxygen consumption rate values in the presence of an ATP synthase inhibitor. · With proper uncoupler titrations, BAM15 offers more flexibility compared to FCCP when performing optimization experiments with valuable samples. Cayman Chemical · (800) 364-9897 www.caymanchem.com 1180 E. Ellsworth Road · Ann Arbor, MI · 48108 Introduction An uncoupler, when used in the context of mitochondrial biology, is any substance that “uncouples” oxidative phosphorylation, or more specifically, the electron transport chain (ETC) from ATP synthase. Uncoupling is accomplished through the dissipation of the mitochondrial membrane potential (ΔψM) without the generation of ATP, resulting in either full or partial mitochondrial depolarization. The loss of membrane potential resulting from uncoupling leads to the generation of heat. Uncouplers exist as proteins (UCPs) found in brown adipose tissue, and as pharmacologic agents such as the classical uncouplers carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP), carbonyl cyanide 3-chlorophenylhydrazone (CCCP), and 2,4-dinitrophenol (DNP). The experimental use of classical uncouplers has played an integral role in determining mitochondrial function, yet they can be difficult to use due to variability between systems and cell lines. For example, FCCP has been increasingly used in cell systems as a means of determining maximal oxygen consumption rate (OCR). This works well, provided that the FCCP concentration has been optimized for the specific cell line, as the effective dose range of FCCP is typically very narrow.
    [Show full text]
  • Seahorse XF Cell Mito Stress Test Kit User Guide 3 4 Agilent Seahorse XF Cell Mito Stress Test Kit User Guide Agilent Seahorse XF Cell Mito Stress Test Kit User Guide
    Agilent Seahorse XF Cell Mito Stress Test Kit User Guide Kit 103015-100 Agilent Technologies Notices © Agilent Technologies, Inc. 2019 Warranty (June 1987) or DFAR 252.227-7015 (b)(2) (November 1995), as applicable in any No part of this manual may be reproduced The material contained in this docu- technical data. in any form or by any means (including ment is provided “as is,” and is sub- electronic storage and retrieval or transla- ject to being changed, without notice, tion into a foreign language) without prior Safety Notices agreement and written consent from in future editions. Further, to the max- Agilent Technologies, Inc. as governed by imum extent permitted by applicable United States and international copyright law, Agilent disclaims all warranties, CAUTION laws. either express or implied, with regard to this manual and any information A CAUTION notice denotes a contained herein, including but not hazard. It calls attention to an oper- Manual Part Number limited to the implied warranties of ating procedure, practice, or the merchantability and fitness for a par- like that, if not correctly performed 103016-400 ticular purpose. Agilent shall not be or adhered to, could result in liable for errors or for incidental or damage to the product or loss of Kit Part Number consequential damages in connection important data. Do not proceed 103015-100 with the furnishing, use, or perfor- beyond a CAUTION notice until the mance of this document or of any indicated conditions are fully Edition information contained herein. Should understood and met. Agilent and the user have a separate Second edition, May 2019 written agreement with warranty Revision G0 terms covering the material in this WARNING Printed in USA document that conflict with these terms, the warranty terms in the sep- A WARNING notice denotes a Agilent Technologies, Inc.
    [Show full text]
  • ATP Synthase: Structure,Abstract: Functionlet F Denote a Eld and and Let Inhibitionv Denote a Vector Space Over F with Nite Positive Dimension
    Spec. Matrices 2019; 7:1–19 Research Article Open Access Kazumasa Nomura* and Paul Terwilliger BioMol Concepts 2019; 10: 1–10 Self-dual Leonard pairs Research Article Open Access https://doi.org/10.1515/spma-2019-0001 Prashant Neupane*, Sudina Bhuju, Nita Thapa,Received Hitesh May 8, 2018; Kumar accepted Bhattarai September 22, 2018 ATP Synthase: Structure,Abstract: FunctionLet F denote a eld and and let InhibitionV denote a vector space over F with nite positive dimension. Consider a pair A, A∗ of diagonalizable F-linear maps on V, each of which acts on an eigenbasis for the other one in an irreducible tridiagonal fashion. Such a pair is called a Leonard pair. We consider the self-dual case in which https://doi.org/10.1515/bmc-2019-0001 there exists an automorphism of the endomorphism algebra of V that swaps A and A∗. Such an automorphism phosphate (Pi), along with considerable release of energy. received September 18, 2018; accepted December 21, 2018. is unique, and called the duality A A∗. In the present paper we give a comprehensive description of this ADP can absorb energy and regain↔ the group to regenerate duality. In particular, we display an invertible F-linear map T on V such that the map X TXT− is the duality Abstract: Oxidative phosphorylation is carried out by an ATP molecule to maintain constant ATP concentration. → A A∗. We express T as a polynomial in A and A∗. We describe how T acts on ags, decompositions, five complexes, which are the sites for electron transport↔ Other than supporting almost all the cellular and ATP synthesis.
    [Show full text]
  • Mitopathways: Respiratory States and Coupling Control Ratios
    O2k-Protocols Mitochondrial Physiology Network 12.15: 1-11 (2011) 2007-2011 OROBOROS Version 6: 2011-12-11 MitoPathways: LEAK OXPHOS ETS Respiratory L/P P P/E States and L ADP uncoupler E Coupling L/E Control Ratios Erich Gnaiger Medical University of Innsbruck D. Swarovski Research Laboratory 6020 Innsbruck, Austria OROBOROS INSTRUMENTS Corp, high-resolution respirometry Schöpfstr 18, A-6020 Innsbruck, Austria [email protected]; www.oroboros.at ‘The growth of any discipline depends on the ability to communicate and develop ideas, and this in turn relies on a language which is sufficiently detailed and flexible.’ Simon Singh (1997) Fermat’s last theorem. Fourth Estate, London. Section 1. Coupling in Oxidative Phosphorylation ................. 1 Page 2. Respiratory Steady-States .................................. 2 2.1. Leak Respiration, L ............................................ 4 2.2. OXPHOS Capacity, P .......................................... 4 2.3. Electron Transfer System Capacity, E................... 5 2.4. Respiratory States of the Intact Cell .................... 5 3. Coupling Control Ratios (CCR), Respiratory Control Ratio (RCR), Uncoupling Control Ratio (UCR) ... 6 4. Summary of Respiratory States .......................... 7 4.1. Residual Oxygen Consumption - ROX ................... 7 4.2. ETS Capacity - State E ....................................... 8 4.3. OXPHOS Capacity – State P ................................ 8 4.4. ROUTINE Respiration – State R ........................... 8 4.5. LEAK Respiration – State
    [Show full text]
  • A Mitochondrial Polymorphism Alters Immune Cell Metabolism and Protects Mice from Skin Inflammation
    International Journal of Molecular Sciences Article A Mitochondrial Polymorphism Alters Immune Cell Metabolism and Protects Mice from Skin Inflammation Paul Schilf 1 , Axel Künstner 1,2 , Michael Olbrich 1, Silvio Waschina 3 , Beate Fuchs 4, Christina E. Galuska 4, Anne Braun 5, Kerstin Neuschütz 1 , Malte Seutter 5, Katja Bieber 1 , Lars Hellberg 6, Christian Sina 7, Tamás Laskay 6, Jan Rupp 6, Ralf J. Ludwig 1,8, Detlef Zillikens 5,8, Hauke Busch 1,2,8 , Christian D. Sadik 5,8 , Misa Hirose 1,8,* and Saleh M. Ibrahim 1,8,9,* 1 Luebeck Institute of Experimental Dermatology, University of Luebeck, 23562 Luebeck, Germany; [email protected] (P.S.); [email protected] (A.K.); [email protected] (M.O.); [email protected] (K.N.); [email protected] (K.B.); [email protected] (R.J.L.); [email protected] (H.B.) 2 Institute of Cardiogenetics, University of Luebeck, 23562 Luebeck, Germany 3 Institute of Human Nutrition and Food Science, Christian-Albrechts-University of Kiel, 24098 Kiel, Germany; [email protected] 4 Leibniz-Institute for Farm Animal Biology (FBN), Core Facility Metabolomics, 18196 Dummerstorf, Germany; [email protected] (B.F.); [email protected] (C.E.G.) 5 Department of Dermatology, University of Luebeck, 23562 Luebeck, Germany; [email protected] (A.B.); [email protected] (M.S.); [email protected] (D.Z.); [email protected] (C.D.S.) 6 Department of Infectious Diseases and Microbiology, University of Luebeck, 23562 Luebeck,
    [Show full text]
  • Biochemical Phenotypes Associated with the Mitochondrial ATP6 Gene Mutations at Nt8993
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Biochimica et Biophysica Acta 1767 (2007) 913–919 www.elsevier.com/locate/bbabio Biochemical phenotypes associated with the mitochondrial ATP6 gene mutations at nt8993 Alessandra Baracca a, Gianluca Sgarbi a, Marina Mattiazzi b, Gabriella Casalena a, Eleonora Pagnotta a, Maria L. Valentino b, Maurizio Moggio c, Giorgio Lenaz a, ⁎ Valerio Carelli b, Giancarlo Solaini a, a Dipartimento di Biochimica “G. Moruzzi”, via Irnerio 48, Università di Bologna, 40126 Bologna, Italy b Dipartimento di Scienze Neurologiche, via U. Foscolo 7, Università di Bologna, Italy c Fondazione Ospedale Maggiore IRCCS-Centro Dino Ferrari, Università di Milano, Italy Received 20 November 2006; received in revised form 7 May 2007; accepted 11 May 2007 Available online 18 May 2007 Abstract Two point mutations (TNG and TNC) at the same 8993 nucleotide of mitochondrial DNA (at comparable mutant load), affecting the ATPase 6 subunit of the F1F0-ATPase, result in neurological phenotypes of variable severity in humans. We have investigated mitochondrial function in lymphocytes from individuals carrying the 8993TNC mutation: the results were compared with data from five 8993TNG NARP (Neuropathy, Ataxia and Retinitis Pigmentosa) patients. Both 8993TNG and 8993TNC mutations led to energy deprivation and ROS overproduction. However, the relative contribution of the two pathogenic components is different depending on the mutation considered. The 8993TNG change mainly induces an energy deficiency, whereas the 8993TNC favours an increased ROS production. These results possibly highlight the different pathogenic mechanism generated by the two mutations at position 8993 and provide useful information to better characterize the biochemical role of the highly conserved Leu-156 in ATPase 6 subunit of the mitochondrial ATP synthase complex.
    [Show full text]
  • Of Human Mitochondrial ATP6 Gene and Its Functional Consequences on Yeast ATP Synthase
    life Article Case Report: Identification of a Novel Variant (m.8909T>C) of Human Mitochondrial ATP6 Gene and Its Functional Consequences on Yeast ATP Synthase 1, 2, 1, 3 1 Qiuju Ding y,Ró˙zaKucharczyk y , Weiwei Zhao y, Alain Dautant , Shutian Xu , Katarzyna Niedzwiecka 2 , Xin Su 3, Marie-France Giraud 3, Kewin Gombeau 3, Mingchao Zhang 1, Honglang Xie 1, Caihong Zeng 1, Marine Bouhier 3, Jean-Paul di Rago 3, Zhihong Liu 1,Déborah Tribouillard-Tanvier 3,4,* and Huimei Chen 1,* 1 National Clinical Research Center of Kidney Diseases, Jinling Hospital, Nanjing University School of Medicine, Nanjing 211166, China; [email protected] (Q.D.); [email protected] (W.Z.); [email protected] (S.X.); [email protected] (M.Z.); [email protected] (H.X.); [email protected] (C.Z.); [email protected] (Z.L.) 2 Institute of Biochemistry and Biophysics, Polish Academy of Sciences, 00090 Warsaw, Poland; [email protected] (R.K.); [email protected] (K.N.) 3 Institut de Biochimie et Génétique Cellulaires, Université de Bordeaux, CNRS, UMR 5095, F-33000 Bordeaux, France; [email protected] (A.D.); [email protected] (X.S.); [email protected] (M.-F.G.); [email protected] (K.G.); [email protected] (M.B.); [email protected] (J.-P.d.R.) 4 Institut national de la santé et de la recherche médicale, 75000 Paris, France * Correspondence: [email protected] (D.T.-T.); [email protected] (H.C.) These authors contributed to this work equally.
    [Show full text]
  • The ATP Content of Carrot Slices Decreased by 20-25
    SALT ACCUMULATION AND ADENOSINE TRIPHOSPHATE IN CARROT XYLEM TISSUE BY M. R. ATKINSON,* GAIL ECKERMANN,* MARY GRANT, AND R. N. ROBERTSON DEPARTMENTS OF AGRICULTURAL BIOCHEMISTRY AND OF BOTANY, THE UNIVERSITY OF ADELAIDE, SOUTH AUSTRALIA Communicated January 3, 1966 Accumulation of potassium or sodium chloride or other metal halides by washed slices of xylem parenchyma from carrot storage roots is associated with increased respiration-the salt respiration. Both salt accumulation and salt respiration are sensitive to inhibitors that block the energy-linked functions of mitochondria, e.g., carbon monoxide in the dark, cyanide, and 2,4-dinitrophenol (for reviews see Robertson' and Briggs, Hope, and Robertson2). It is not known if accumulation of salts by plant tissues is directly coupled to the electron and hydrogen transport of salt respiration or if it is dependent on the hydrolysis of adenosine triphosphate (ATP) formed by the salt respiration. It has been postulated1 that if salt ac- cumulation is directly coupled, it might be an alternative to oxidative phosphoryla- tion. This paper reports some experiments which might distinguish between these two possibilities. A method for analysis of ATP in small samples of carrot discs has now been developed, and the effects of metabolic inhibitors on salt accumulation, salt res- piration, and ATP concentration have been studied. Experimental.-Carrot slices were prepared by cutting and washing with frequent changes of deionized water in the first few hours. Subsequently they were kept in aerated deionized water or chloromycetin' (50 ,ug/ml; for details see Table 1) changed daily for 7 days before use. Although the conditions of washing do not favor bacterial growth, it was found by direct counting and by dilution in nutrient agar that slices washed in water were contaminated with 107 to 108 bacteria/gm wet weight of tissue.
    [Show full text]