DOCSLIB.ORG

ALS-Linked Mutant Superoxide Dismutase 1 (SOD1) Alters Mitochondrial Protein Composition and Decreases Protein Import

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
ALS-Linked Mutant Superoxide Dismutase 1 (SOD1) Alters Mitochondrial Protein Composition and Decreases Protein Import ALS-linked mutant superoxide dismutase 1 (SOD1) alters mitochondrial protein composition and decreases protein import Quan Lia,1, Christine Vande Veldeb,c,1, Adrian Israelsonb, Jing Xied, Aaron O. Baileye, Meng-Qui Donge, Seung-Joo Chuna, Tamal Royb, Leah Winera, John R. Yatese, Roderick A. Capaldid, Don W. Clevelandb,2, and Timothy M. Millera,b,2 aDepartment of Neurology, Hope Center for Neurological Disorders, The Washington University School of Medicine, St. Louis, MO 63110; bLudwig Institute and Departments of Cellular and Molecular Medicine and Neuroscience, University of California at San Diego, La Jolla, CA 92093-0670; cDepartment of Medicine, Centre Hospitalier de l’Université de Montréal Research Center, Université de Montréal, Montreal, QC, Canada H2L 4M1; eDepartment of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037; and dMitoSciences, Eugene, OR 97403 Contributed by Don W. Cleveland, October 18, 2010 (sent for review February 4, 2010) Mutations in superoxide dismutase 1 (SOD1) cause familial ALS. system very little SOD1 is found in or on mitochondria (20, 21), Mutant SOD1 preferentially associates with the cytoplasmic face of but mutant SOD1 has been found associated presymptomatically mitochondria from spinal cords of rats and mice expressing SOD1 with the cytoplasmic face of mitochondria from spinal cord in all mutations. Two-dimensional gels and multidimensional liquid chro- rodent models of SOD1 mutant-mediated disease (20, 21). A matography, in combination with tandem mass spectrometry, criticism that apparent association of dismutase inactive mutants fl revealed 33 proteins that were increased and 21 proteins that with mitochondria might re ect cosedimentation of protein G93A aggregates rather than bona fide mitochondrial association (22) were decreased in SOD1 rat spinal cord mitochondria com- fl pared with SOD1WT spinal cord mitochondria. Analysis of this has been refuted by demonstrating that mutant SOD1 oats with group of proteins revealed a higher-than-expected proportion in- mitochondria rather than sedimenting as much more dense pro- tein-only aggregates would under such conditions (21). Indeed, volved in complex I and protein import pathways. Direct import a portion of misfolded mutant SOD1 directly binds to the cyto- assays revealed a 30% decrease in protein import only in spinal plasmic facing surface of the major voltage-dependent anion cord mitochondria, despite an increase in the mitochondrial import NEUROSCIENCE G93A channel (VDAC1), a general diffusion pore for anions and cati- components TOM20, TOM22, and TOM40. Recombinant SOD1 ons that is located on the outer mitochondrial membrane. Mutant G85R WT ’ or SOD1 , but not SOD1 or a Parkinson s disease-causing, mis- SOD1 binding reduces conductance by purified VDAC1 recon- α E46K > folded -synuclein mutant, decreased protein import by 50% stituted in a lipid bilayer, and spinal cord mitochondria have re- in nontransgenic mitochondria from spinal cord, but not from liver. duced ADP uptake (23). Thus, altered mitochondrial protein content accompanied by selec- Three prior efforts have reported use of proteomic tools to in- tive decreases in protein import into spinal cord mitochondria com- vestigate how mutant SOD1 affects mitochondrial protein com- prises part of the mitochondrial damage arising from mutant SOD1. position within whole-spinal-cord extracts of dismutase-active (24) or dismutase-inactive (25) SOD1 mutant mouse models or, more motor neuron disease | proteomic | neurodegenerative disease | selectively, within mitochondria of a cell line retaining some motor amyotrophic neuron-like properties and expressing one dismutase-active, ALS- linked SOD1 mutant (26). We extended these approaches to de- myotrophic lateral sclerosis (ALS) is an adult-onset neuro- termine whether mutant SOD1 affects protein content of spinal Adegenerative disorder caused by loss of the motor neurons cord mitochondria before disease initiation in rodent models that within the spinal cord and dysfunction of the motor pathways in the develop ALS-like disease. Using a combination of mass spectrom- cortex (1, 2). Most cases of ALS (90%) are sporadic. Of the 10% of etry and 2D gels, we now report 21 proteins to be decreased ap- cases that are inherited, about one-fifth are caused by mutation proximately twofold, with another 33 that are increased by a within the superoxide dismutase 1 (SOD1) gene. Rodents expres- comparable amount or more in mitochondria isolated from spinal cords of mutant SOD1 animals, accompanied by diminished pro- sing mutant SOD1 develop an ALS-like disease (1, 2). A consensus fi has emerged that disease arises from acquisition by the mutant tein import activity. By using puri ed components, inhibition of proteins of one or more toxic properties, rather than from loss of mitochondrial protein import is demonstrated to be a direct result dismutase activity (3, 4). At least eight prominent mutant-derived of mutant but not wild-type SOD1. toxicities have been proposed, including mitochondrial dysfunc- Results tion, excitotoxicity, endoplasmic reticulum stress, toxic extracel- lular SOD1, superoxide production from microglia or astrocytes, Altered Spinal Cord Mitochondrial Protein Content in SOD1 Mutant and disruption of the blood–brain barrier (5). Furthermore, Rats. To identify possible differences in protein composition be- pathogenesis is noncell autonomous with disease progression tween mitochondria isolated from spinal cords of age-matched driven by mutant synthesis in astrocytes (6–8) and microglia (3, 9). SOD1WT and SOD1G93A mutant rats, we used 2D gel electro- Dysfunction of mitochondria can clearly cause specific damage phoresis as well as liquid chromatography followed by tandem to neurons or muscle because deletion/mutation in mitochondrial mass spectrometry [multidimensional protein identification tech- genes causes specific nerve and muscle diseases (10). Accompa- nying mutant SOD1-mediated disease in some mice is evidence for altered mitochondrial calcium-buffering capacity (11) and Author contributions: Q.L., C.V.V., R.A.C., D.W.C., and T.M.M. designed research; Q.L., C.V.V., changes in the activity of complexes of the electron transport chain A.I., J.X., A.O.B., M.-Q.D., S.-J.C., T.R., L.W., and T.M.M. performed research; Q.L., C.V.V., A.I., (12). Deletion of the mitochondrial BCL-2–related proteins BAX J.X., A.O.B., M.-Q.D., S.-J.C., T.R., L.W., J.R.Y., R.A.C., D.W.C., and T.M.M. analyzed data; and and BAK delays disease onset in SOD1G93A mice (13), perhaps Q.L., C.V.V., D.W.C., and T.M.M. wrote the paper. by modulating effects of SOD1 conformational changes to BCL-2 The authors declare no conflict of interest. (14). Apparent damage to mitochondria has been reported in 1Q.L. and C.V.V. contributed equally to this work. autopsy material from ALS patients (15, 16). 2To whom correspondence may be addressed. E-mail: [email protected] or Although SOD1 is a primarily cytosolic enzyme, wild-type [email protected]. SOD1 is also found in the intermembrane space of mitochondria This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. in liver (17) and in yeast (18, 19). However, in the normal nervous 1073/pnas.1014862107/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1014862107 PNAS Early Edition | 1of6 Downloaded by guest on September 25, 2021 G93A nology (MudPIT)] (27). Initially, a presymptomatic age (11 wk) Decreased Complex I Activity in Mitochondria from SOD1 Rats. was chosen because it was known to correspond to a time of Six (NDUFS1, ND5, NDUFB8, NDUFC2, NDUFA5, NDUFB9) very active denervation of lower motor neurons (28). Although of the 26 down-regulated proteins in mitochondria from mutant a change in staining on 2D gels may reflect altered abundance, spinal cord are components of the 42-subunit complex I, also size, or isoelectric point, the overall patterns of major proteins known as NADH dehydrogenase. Complex I is located on the (visible in Coomassie (Fig. 1) or silver (Fig. S1) staining were inner membrane of mitochondria, catalyzes the shuttling of elec- highly similar. Twelve spots (Fig. 1 B and C compared with Fig. 1 trons from NADH to coenzyme Q, and helps to build the proton E and F and Fig. S1 B and C compared with Fig. S1 E and F) were gradient required for ATP generation. Immunoprecipitation of clearly different between the two samples, with five (spots 1, 6, 7, mitochondrial extracts with a monoclonal antibody raised against 8, and 9) increased in SOD1G93A mitochondria and seven (spots complex I, which had been crosslinked to protein G agarose beads (32), confirmed that at least six complex I polypeptides were 2–5 and 10–12) decreased. Each was excised, digested with tryp- changed in relative abundance: two were increased [(V1, S2) and sin, and identified via tandem mass spectrometry (Table S1). (A10, ND5)] and four were decreased [S1, (A9, D1), S3, and (B10, Three spots contained more than one protein, resulting in a total fi V2; B7, A5, B4, B5, B6, S5, B16, 6, A7)] (some stained bands of 15 proteins identi ed. represent multiple polypeptides) (Fig. 3A). Of those decreased in Using MudPIT mass spectrometry, protein differences were G93A fi SOD1 mitochondria, a polypeptide migrating at 75 kDa (Fig. also identi ed from three independent preparations of mitochon- 3A) most likely represents S1 (also known as NDUFS1) (32–34), dria from 11-wk-old SOD1WT and age-matched presymptomatic G93A which was also seen to be decreased by mass spectrometry (Table SOD1 rat spinal cords, using normalized spectral counts as an 1) and 2D gels (Fig. 3B, arrows). A prominent complex I poly- indication of the relative amount of any given protein (29, 30). peptide migrating at 35 kDa, most likely representing ND5 (32– We identified 299 mitochondrial proteins present in all of the 34), was increased in immunoprecipitates. ND5 is a mitochond- mitochondrial samples.
Load more

Recommended publications
  • Supplementary Materials: Evaluation of Cytotoxicity and Α-Glucosidase Inhibitory Activity of Amide and Polyamino-Derivatives of Lupane Triterpenoids
    Supplementary Materials: Evaluation of cytotoxicity and α-glucosidase inhibitory activity of amide and polyamino-derivatives of lupane triterpenoids Oxana B. Kazakova1*, Gul'nara V. Giniyatullina1, Akhat G. Mustafin1, Denis A. Babkov2, Elena V. Sokolova2, Alexander A. Spasov2* 1Ufa Institute of Chemistry of the Ufa Federal Research Centre of the Russian Academy of Sciences, 71, pr. Oktyabrya, 450054 Ufa, Russian Federation 2Scientific Center for Innovative Drugs, Volgograd State Medical University, Novorossiyskaya st. 39, Volgograd 400087, Russian Federation Correspondence Prof. Dr. Oxana B. Kazakova Ufa Institute of Chemistry of the Ufa Federal Research Centre of the Russian Academy of Sciences 71 Prospeсt Oktyabrya Ufa, 450054 Russian Federation E-mail: [email protected] Prof. Dr. Alexander A. Spasov Scientific Center for Innovative Drugs of the Volgograd State Medical University 39 Novorossiyskaya st. Volgograd, 400087 Russian Federation E-mail: [email protected] Figure S1. 1H and 13C of compound 2. H NH N H O H O H 2 2 Figure S2. 1H and 13C of compound 4. NH2 O H O H CH3 O O H H3C O H 4 3 Figure S3. Anticancer screening data of compound 2 at single dose assay 4 Figure S4. Anticancer screening data of compound 7 at single dose assay 5 Figure S5. Anticancer screening data of compound 8 at single dose assay 6 Figure S6. Anticancer screening data of compound 9 at single dose assay 7 Figure S7. Anticancer screening data of compound 12 at single dose assay 8 Figure S8. Anticancer screening data of compound 13 at single dose assay 9 Figure S9. Anticancer screening data of compound 14 at single dose assay 10 Figure S10.
    [Show full text]
  • Location Analysis of Estrogen Receptor Target Promoters Reveals That
    Location analysis of estrogen receptor ␣ target promoters reveals that FOXA1 defines a domain of the estrogen response Jose´ e Laganie` re*†, Genevie` ve Deblois*, Ce´ line Lefebvre*, Alain R. Bataille‡, Franc¸ois Robert‡, and Vincent Gigue` re*†§ *Molecular Oncology Group, Departments of Medicine and Oncology, McGill University Health Centre, Montreal, QC, Canada H3A 1A1; †Department of Biochemistry, McGill University, Montreal, QC, Canada H3G 1Y6; and ‡Laboratory of Chromatin and Genomic Expression, Institut de Recherches Cliniques de Montre´al, Montreal, QC, Canada H2W 1R7 Communicated by Ronald M. Evans, The Salk Institute for Biological Studies, La Jolla, CA, July 1, 2005 (received for review June 3, 2005) Nuclear receptors can activate diverse biological pathways within general absence of large scale functional data linking these putative a target cell in response to their cognate ligands, but how this binding sites with gene expression in specific cell types. compartmentalization is achieved at the level of gene regulation is Recently, chromatin immunoprecipitation (ChIP) has been used poorly understood. We used a genome-wide analysis of promoter in combination with promoter or genomic DNA microarrays to occupancy by the estrogen receptor ␣ (ER␣) in MCF-7 cells to identify loci recognized by transcription factors in a genome-wide investigate the molecular mechanisms underlying the action of manner in mammalian cells (20–24). This technology, termed 17␤-estradiol (E2) in controlling the growth of breast cancer cells. ChIP-on-chip or location analysis, can therefore be used to deter- We identified 153 promoters bound by ER␣ in the presence of E2. mine the global gene expression program that characterize the Motif-finding algorithms demonstrated that the estrogen re- action of a nuclear receptor in response to its natural ligand.
    [Show full text]
  • Assembly Factors for the Membrane Arm of Human Complex I
    Assembly factors for the membrane arm of human complex I Byron Andrews, Joe Carroll, Shujing Ding, Ian M. Fearnley, and John E. Walker1 Medical Research Council Mitochondrial Biology Unit, Cambridge CB2 0XY, United Kingdom Contributed by John E. Walker, October 14, 2013 (sent for review September 12, 2013) Mitochondrial respiratory complex I is a product of both the nuclear subunits in a fungal enzyme from Yarrowia lipolytica seem to be and mitochondrial genomes. The integration of seven subunits distributed similarly (12, 13). encoded in mitochondrial DNA into the inner membrane, their asso- The assembly of mitochondrial complex I involves building the ciation with 14 nuclear-encoded membrane subunits, the construc- 44 subunits emanating from two genomes into the two domains of tion of the extrinsic arm from 23 additional nuclear-encoded the complex. The enzyme is put together from preassembled sub- proteins, iron–sulfur clusters, and flavin mononucleotide cofactor complexes, and their subunit compositions have been characterized require the participation of assembly factors. Some are intrinsic to partially (14, 15). Extrinsic assembly factors of unknown function the complex, whereas others participate transiently. The suppres- become associated with subcomplexes that accumulate when as- sion of the expression of the NDUFA11 subunit of complex I dis- sembly and the activity of complex I are impaired by pathogenic rupted the assembly of the complex, and subcomplexes with mutations. Some assembly factor mutations also impair its activ- masses of 550 and 815 kDa accumulated. Eight of the known ex- ity (16). Other pathogenic mutations are found in all of the core trinsic assembly factors plus a hydrophobic protein, C3orf1, were subunits, and in 10 supernumerary subunits (NDUFA1, NDUFA2, associated with the subcomplexes.
    [Show full text]
  • NDUFS6 Mutations Are a Novel Cause of Lethal Neonatal Mitochondrial Complex I Deficiency Denise M
    Related Commentary, page 760 Research article NDUFS6 mutations are a novel cause of lethal neonatal mitochondrial complex I deficiency Denise M. Kirby,1,2,3 Renato Salemi,1 Canny Sugiana,1,3 Akira Ohtake,4 Lee Parry,1 Katrina M. Bell,1 Edwin P. Kirk,5 Avihu Boneh,1,2,3 Robert W. Taylor,6 Hans-Henrik M. Dahl,1,3 Michael T. Ryan,4 and David R. Thorburn1,2,3 1Murdoch Childrens Research Institute and 2Genetic Health Services Victoria, Royal Children’s Hospital, Melbourne, Victoria, Australia. 3Department of Paediatrics, University of Melbourne, Melbourne, Victoria, Australia. 4Department of Biochemistry, LaTrobe University, Melbourne, Victoria, Australia. 5Department of Medical Genetics, Sydney Children’s Hospital, Sydney, New South Wales, Australia. 6Mitochondrial Research Group, School of Neurology, Neurobiology and Psychiatry, University of Newcastle upon Tyne, Newcastle upon Tyne, United Kingdom. Complex I deficiency, the most common respiratory chain defect, is genetically heterogeneous: mutations in 8 nuclear and 7 mitochondrial DNA genes encoding complex I subunits have been described. However, these genes account for disease in only a minority of complex I–deficient patients. We investigated whether there may be an unknown common gene by performing functional complementation analysis of cell lines from 10 unrelated patients. Two of the patients were found to have mitochondrial DNA mutations. The other 8 repre- sented 7 different (nuclear) complementation groups, all but 1 of which showed abnormalities of complex I assembly. It is thus unlikely that any one unknown gene accounts for a large proportion of complex I cases. The 2 patients sharing a nuclear complementation group had a similar abnormal complex I assembly profile and were studied further by homozygosity mapping, chromosome transfers, and microarray expression analysis.
    [Show full text]
  • Proteomic and Metabolomic Analyses of Mitochondrial Complex I-Deficient
    THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 287, NO. 24, pp. 20652–20663, June 8, 2012 © 2012 by The American Society for Biochemistry and Molecular Biology, Inc. Published in the U.S.A. Proteomic and Metabolomic Analyses of Mitochondrial Complex I-deficient Mouse Model Generated by Spontaneous B2 Short Interspersed Nuclear Element (SINE) Insertion into NADH Dehydrogenase (Ubiquinone) Fe-S Protein 4 (Ndufs4) Gene*□S Received for publication, November 25, 2011, and in revised form, April 5, 2012 Published, JBC Papers in Press, April 25, 2012, DOI 10.1074/jbc.M111.327601 Dillon W. Leong,a1 Jasper C. Komen,b1 Chelsee A. Hewitt,a Estelle Arnaud,c Matthew McKenzie,d Belinda Phipson,e Melanie Bahlo,e,f Adrienne Laskowski,b Sarah A. Kinkel,a,g,h Gayle M. Davey,g William R. Heath,g Anne K. Voss,a,h René P. Zahedi,i James J. Pitt,j Roman Chrast,c Albert Sickmann,i,k Michael T. Ryan,l Gordon K. Smyth,e,f,h b2 a,h,m,n3 David R. Thorburn, and Hamish S. Scott Downloaded from From the aMolecular Medicine Division, gImmunology Division, and eBioinformatics Division, Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia, the bMurdoch Childrens Research Institute, Royal Children’s Hospital and Department of Paediatrics, University of Melbourne, Parkville, Victoria 3052, Australia, the cDépartement de Génétique Médicale, Université de Lausanne, 1005 Lausanne, Switzerland, the dCentre for Reproduction and Development, Monash Institute of Medical Research, Clayton, Victoria 3168, Australia, the hDepartment of Medical Biology
    [Show full text]
  • S41467-020-18249-3.Pdf
    ARTICLE https://doi.org/10.1038/s41467-020-18249-3 OPEN Pharmacologically reversible zonation-dependent endothelial cell transcriptomic changes with neurodegenerative disease associations in the aged brain Lei Zhao1,2,17, Zhongqi Li 1,2,17, Joaquim S. L. Vong2,3,17, Xinyi Chen1,2, Hei-Ming Lai1,2,4,5,6, Leo Y. C. Yan1,2, Junzhe Huang1,2, Samuel K. H. Sy1,2,7, Xiaoyu Tian 8, Yu Huang 8, Ho Yin Edwin Chan5,9, Hon-Cheong So6,8, ✉ ✉ Wai-Lung Ng 10, Yamei Tang11, Wei-Jye Lin12,13, Vincent C. T. Mok1,5,6,14,15 &HoKo 1,2,4,5,6,8,14,16 1234567890():,; The molecular signatures of cells in the brain have been revealed in unprecedented detail, yet the ageing-associated genome-wide expression changes that may contribute to neurovas- cular dysfunction in neurodegenerative diseases remain elusive. Here, we report zonation- dependent transcriptomic changes in aged mouse brain endothelial cells (ECs), which pro- minently implicate altered immune/cytokine signaling in ECs of all vascular segments, and functional changes impacting the blood–brain barrier (BBB) and glucose/energy metabolism especially in capillary ECs (capECs). An overrepresentation of Alzheimer disease (AD) GWAS genes is evident among the human orthologs of the differentially expressed genes of aged capECs, while comparative analysis revealed a subset of concordantly downregulated, functionally important genes in human AD brains. Treatment with exenatide, a glucagon-like peptide-1 receptor agonist, strongly reverses aged mouse brain EC transcriptomic changes and BBB leakage, with associated attenuation of microglial priming. We thus revealed tran- scriptomic alterations underlying brain EC ageing that are complex yet pharmacologically reversible.
    [Show full text]
  • Low Abundance of the Matrix Arm of Complex I in Mitochondria Predicts Longevity in Mice
    ARTICLE Received 24 Jan 2014 | Accepted 9 Apr 2014 | Published 12 May 2014 DOI: 10.1038/ncomms4837 OPEN Low abundance of the matrix arm of complex I in mitochondria predicts longevity in mice Satomi Miwa1, Howsun Jow2, Karen Baty3, Amy Johnson1, Rafal Czapiewski1, Gabriele Saretzki1, Achim Treumann3 & Thomas von Zglinicki1 Mitochondrial function is an important determinant of the ageing process; however, the mitochondrial properties that enable longevity are not well understood. Here we show that optimal assembly of mitochondrial complex I predicts longevity in mice. Using an unbiased high-coverage high-confidence approach, we demonstrate that electron transport chain proteins, especially the matrix arm subunits of complex I, are decreased in young long-living mice, which is associated with improved complex I assembly, higher complex I-linked state 3 oxygen consumption rates and decreased superoxide production, whereas the opposite is seen in old mice. Disruption of complex I assembly reduces oxidative metabolism with concomitant increase in mitochondrial superoxide production. This is rescued by knockdown of the mitochondrial chaperone, prohibitin. Disrupted complex I assembly causes premature senescence in primary cells. We propose that lower abundance of free catalytic complex I components supports complex I assembly, efficacy of substrate utilization and minimal ROS production, enabling enhanced longevity. 1 Institute for Ageing and Health, Newcastle University, Newcastle upon Tyne NE4 5PL, UK. 2 Centre for Integrated Systems Biology of Ageing and Nutrition, Newcastle University, Newcastle upon Tyne NE4 5PL, UK. 3 Newcastle University Protein and Proteome Analysis, Devonshire Building, Devonshire Terrace, Newcastle upon Tyne NE1 7RU, UK. Correspondence and requests for materials should be addressed to T.v.Z.
    [Show full text]
  • Membrane Proteomics of Cervical Cancer Cell Lines Reveal Insights on the Process of Cervical Carcinogenesis
    INTERNATIONAL JOURNAL OF ONCOLOGY 53: 2111-2122, 2018 Membrane proteomics of cervical cancer cell lines reveal insights on the process of cervical carcinogenesis KALLIOPI I. PAPPA1,2, POLYXENI CHRISTOU3,4, AMARILDO XHOLI3, GEORGE MERMELEKAS3, GEORGIA KONTOSTATHI3,4, VASILIKI LYGIROU3,4, MANOUSOS MAKRIDAKIS3, JEROME ZOIDAKIS3 and NICHOLAS P. ANAGNOU1,4 1Cell and Gene Therapy Laboratory, Centre of Basic Research II, Biomedical Research Foundation of the Academy of Athens, 11527 Athens; 2First Department of Obstetrics and Gynecology, University of Athens School of Medicine, Alexandra Hospital, 11528 Athens; 3Biotechnology Division, Centre of Basic Research, Biomedical Research Foundation of the Academy of Athens; 4Laboratory of Biology, University of Athens School of Medicine, 11527 Athens, Greece Received March 22, 2018; Accepted May 4, 2018 DOI: 10.3892/ijo.2018.4518 Abstract. The available therapeutic approaches for cervical biological pathways relevant to malignancy, including ‘HIPPO cancer can seriously affect the fertility potential of patient; signaling’, ‘PI3K/Akt signaling’, ‘cell cycle: G2/M DNA thus, there is a pressing requirement for less toxic and damage checkpoint regulation’ and ‘EIF2 signaling’. These targeted therapies. The membrane proteome is a potential unique membrane protein identifications offer insights on a source of therapeutic targets; however, despite the signifi- previously inaccessible region of the cervical cancer proteome, cance of membrane proteins in cancer, proteomic analysis and may represent putative
    [Show full text]
  • THE FUNCTIONAL SIGNIFICANCE of MITOCHONDRIAL SUPERCOMPLEXES in C. ELEGANS by WICHIT SUTHAMMARAK Submitted in Partial Fulfillment
    THE FUNCTIONAL SIGNIFICANCE OF MITOCHONDRIAL SUPERCOMPLEXES in C. ELEGANS by WICHIT SUTHAMMARAK Submitted in partial fulfillment of the requirements For the degree of Doctor of Philosophy Dissertation Advisor: Drs. Margaret M. Sedensky & Philip G. Morgan Department of Genetics CASE WESTERN RESERVE UNIVERSITY January, 2011 CASE WESTERN RESERVE UNIVERSITY SCHOOL OF GRADUATE STUDIES We hereby approve the thesis/dissertation of _____________________________________________________ candidate for the ______________________degree *. (signed)_______________________________________________ (chair of the committee) ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ (date) _______________________ *We also certify that written approval has been obtained for any proprietary material contained therein. Dedicated to my family, my teachers and all of my beloved ones for their love and support ii ACKNOWLEDGEMENTS My advanced academic journey began 5 years ago on the opposite side of the world. I traveled to the United States from Thailand in search of a better understanding of science so that one day I can return to my homeland and apply the knowledge and experience I have gained to improve the lives of those affected by sickness and disease yet unanswered by science. Ultimately, I hoped to make the academic transition into the scholarly community by proving myself through scientific research and understanding so that I can make a meaningful contribution to both the scientific and medical communities. The following dissertation would not have been possible without the help, support, and guidance of a lot of people both near and far. I wish to thank all who have aided me in one way or another on this long yet rewarding journey. My sincerest thanks and appreciation goes to my advisors Philip Morgan and Margaret Sedensky.
    [Show full text]
  • Analyzing the Function of TRAP1 in Models of Parkinson's Disease
    Analyzing the function of TRAP1 in models of Parkinson’s disease Von der Fakultät für Mathematik, Informatik und Naturwissenschaften der RWTH Aachen University zur Erlangung des akademischen Grades einer Doktorin der Naturwissenschaften genehmigte Dissertation vorgelegt von Li Zhang aus Changchun, Jilin (China) Berichter: Universitätsprofessor Dr. med. Jörg B. Schulz Universitätsprofessor Dr. rer. nat. Marc Spehr Tag der mündlichen Prüfung: 29.01.2016 Diese Dissertation ist auf den Internetseiten der Universitätsbibliothek online verfügbar. Eidesstattliche Versicherung ___________________________Zhang, Li ___________________________ Name, Vorname Matrikelnummer (freiwillige Angabe) Ich versichere hiermit an Eides Statt, dass ich die vorliegende Arbeit/Bachelorarbeit/ Masterarbeit* mit dem Titel __________________________________________________________________________Analyzing the function of TRAP1 in models of Parkinson’s disease __________________________________________________________________________Uebersetzung: Analyse der TRAP1-Funktion in Modellen fuer Morbus Parkinson __________________________________________________________________________ selbständig und ohne unzulässige fremde Hilfe erbracht habe. Ich habe keine anderen als die angegebenen Quellen und Hilfsmittel benutzt. Für den Fall, dass die Arbeit zusätzlich auf einem Datenträger eingereicht wird, erkläre ich, dass die schriftliche und die elektronische Form vollständig übereinstimmen. Die Arbeit hat in gleicher oder ähnlicher Form noch keiner Prüfungsbehörde vorgelegen.
    [Show full text]
  • Polymorphisms in Mitochondrial Genes and Prostate Cancer Risk
    3558 Polymorphisms in Mitochondrial Genes and Prostate Cancer Risk Liang Wang,1 Shannon K. McDonnell,2 Scott J. Hebbring,1 Julie M. Cunningham,1 Jennifer St Sauver,2 James R. Cerhan,2 Grazia Isaya,3 Daniel J. Schaid,2 and Stephen N. Thibodeau1 Departments of 1Laboratory Medicine and Pathology, 2Health Sciences Research, and 3Pediatrics, Mayo Clinic College of Medicine, Rochester, Minnesota Abstract The mitochondrion, conventionally thought to be an cancer after adjusting for multiple testing. Nonetheless, organelle specific to energy metabolism, is in fact we evaluated unadjusted P values by comparing our multifunctional and implicated in many diseases, results with those from the Cancer Genetic Markers of including cancer. To evaluate whether mitochondria- Susceptibility (CGEMS) phase I data set. Seven related genes are associated with increased risk for tagSNPs had unadjusted P V 0.05 in both our data prostate cancer, we genotyped 24 single-nucleotide and in CGEMS (two SNPs were identical and five were polymorphisms (SNP) within the mitochondrial ge- in strong linkage disequilibrium with CGEMS SNPs). nome and 376 tagSNPs localized to 78 nuclear-encoded These seven SNPs (rs17184211, rs4147684, rs4233367, mitochondrial genes. The tagSNPs were selected to rs2070902, rs3829037, rs7830235, and rs1203213) are achieve z80% coverage based on linkage disequilibri- located in genes MTRR, NDUFA9, NDUFS2, NDUFB9, um. We compared allele and haplotype frequencies in and COX7A2, respectively. Five of the seven SNPs were f1,000 prostate cancer cases with f500 population further included in the CGEMS phase II study; controls. An association with prostate cancer was not however, none of the findings for these were replicat- detected for any of the SNPs within the mitochondrial ed.
    [Show full text]
  • Mitochondrial Dysfunction in Parkinson's Disease: Focus on Mitochondrial
    biomedicines Review Mitochondrial Dysfunction in Parkinson’s Disease: Focus on Mitochondrial DNA Olga Buneeva, Valerii Fedchenko, Arthur Kopylov and Alexei Medvedev * Institute of Biomedical Chemistry, 10 Pogodinskaya Street, 119121 Moscow, Russia; [email protected] (O.B.); [email protected] (V.F.); [email protected] (A.K.) * Correspondence: [email protected]; Tel.: +7-495-245-0509 Received: 17 November 2020; Accepted: 8 December 2020; Published: 10 December 2020 Abstract: Mitochondria, the energy stations of the cell, are the only extranuclear organelles, containing their own (mitochondrial) DNA (mtDNA) and the protein synthesizing machinery. The location of mtDNA in close proximity to the oxidative phosphorylation system of the inner mitochondrial membrane, the main source of reactive oxygen species (ROS), is an important factor responsible for its much higher mutation rate than nuclear DNA. Being more vulnerable to damage than nuclear DNA, mtDNA accumulates mutations, crucial for the development of mitochondrial dysfunction playing a key role in the pathogenesis of various diseases. Good evidence exists that some mtDNA mutations are associated with increased risk of Parkinson’s disease (PD), the movement disorder resulted from the degenerative loss of dopaminergic neurons of substantia nigra. Although their direct impact on mitochondrial function/dysfunction needs further investigation, results of various studies performed using cells isolated from PD patients or their mitochondria (cybrids) suggest their functional importance. Studies involving mtDNA mutator mice also demonstrated the importance of mtDNA deletions, which could also originate from abnormalities induced by mutations in nuclear encoded proteins needed for mtDNA replication (e.g., polymerase γ). However, proteomic studies revealed only a few mitochondrial proteins encoded by mtDNA which were downregulated in various PD models.
    [Show full text]