Functional analysis of the apoptosis-inducing factor (AIF) [Elektronische Ressource] / vorgelegt von Nicola Vahsen

Sujets

• Biologie

Informations

• Publié par: universitat_duisburg-essen
• Publié le: 01 janvier 2006
• Nombre de lectures: 55
• Langue: English
• Poids de l'ouvrage: 18 Mo

Extrait

Functional Analysis of the Apoptosis-Inducing Factor
(AIF)
Inaugural-Dissertation
zur
Erlangung des Doktorgrades
Dr. rer. nat.
des Fachbereichs
Biologie und Geographie
an der
Universität Duisburg-Essen
vorgelegt von
Nicola Vahsen
geboren in Mülheim an der Ruhr
Datum der mündlichen Prüfung: 05.April 2006
Gutachter:
Guido Kroemer, Prof.Dr.med.Dr.rer.nat. (Institut Gustave Roussy,Villejuif,Frankreich)
Prof.Dr.Dr. Brigitte Pützer, Prof.Dr.med.Dr.rer.nat. (Universtät Rostock)
Prof.Dr. Helmut Esche, Prof.Dr.rer.nat. (Universität Duisburg-Essen)
IAbbreviations
µg Microgramme
µl Microlitre
ADP Adenosine biphosphate
AIF Apoptosis-inducing factor
AIFL AIF-like protein
ANT Adenine nucleotide transporter
Apaf-1 Apoptosis protease activating factor 1
APS Ammonium persulphate
ATP Adenosine triphosphate
Bcl-2 B-cell leukemia/lymphoma 2
BH Bcl-2 homology
BN Blue native
BSA Bovine serum albumin
BSO Buthionine sulfoximine
C Celsius
C.elegans Caenorhabditis elegans
CAD Caspase-activated DNAse
cDNA Complementary DNA
CHAPS 3-[(3-Cholamidopropyl)-dimethylammonio)]-1-propane
sulfonic acid
CIA30 Complex I assembly factor 30
Cox1/ Cox2 Cytochrome c oxidase subunit 1/2
CPEO Chronic external ophtamoplegia
CPS6 Ced-3 protease suppressor 6
CTL Cytotoxic T lymphocytes
CY Cyclophosmamide
CypA Cyclophilin A
Cyt c Cytochrome c
cytB Cytochrome B
Da Dalton
DAPI 4’,-6-diamino-2-phenylindole-dihydrochloride
DCF-DA 2’7’-dichlorodihydrofluorescein diacetate
DCPIP Dichlorophenol-indophenol
DIABLO Direct IAP-binding protein with low PI
DISC Death inducing signalling complex
DMSO Dimethyl sulfoxide
DNA Deoxyribonucleic acid
dNTP Deoxyribonucleotide triphosphate
dsDNA Double-stranded DNA
dsRNA Double-stranded RNA
DTT Dithiothreitol
EDTA Ethylene diamine tetraacetic acid
EGTA Ethylene-glycol-bis(2-aminoethylether)-N-N-N-N’-tetraacetic
acid
EM Electron microscopy
EndoG Endonuclease G
ER Endoplasmic reticulum
ES cells Embryonic stem cells
IIAbbreviations continued
F Farad
FACS Fluorescence-activated cell sorter
FAD Flavine-adenine dinucleotide
FADD Fas-associated death domain protein
FCS Fetal calf serum
FITC Fluorescein isothiocyanate
g Gramme
GAPDH Glyceraldehyde-3-phosphate deshydrogenase
GFP Green fluorescent protein
GPT Glutamate-pyruvate-transaminase
GSH Glutathione
H DCFDA Dichlorodihydrofluorescein diacetate2
HIV Human immunodeficiency virus
HnRNPA1 Heterogenous nuclear ribonucleoprotein A1
Hsp Heat shock protein
HtrA2 High temperature required protein A2
IAP Inhibtior of apoptosis protein
ICAD Inhibitor of caspase-activated DNAse
Ig Immunoglobulin
IP Immunoprecipitation
KO Knock-out
l Litre
LDH Lactate-dehydrogenase
LIF Leukemia inducing factor
M Molar
mA milli-Amper
MELAS Mitochondrial encephalopathy lactic acidosis syndrome
ml Millilitre
mM Millimolar
MnTBAP Mn(III)tetrakis(4-benzoic acid)porphyrin chloride
MOMP Mitochondrial outer membrane permeabilization
mRNA Messenger RNA
NAD Nicotinamide adenine dinucleotide
NADP Nicotinamide adenine dinucleotide phosphate
NAO Nonyl acridine orange
ND1/ ND6 NADH dehydrogenase subunit 1/6
NDUFA9 NADH-ubiquinone oxydoreductase 1 alpha subcomplex 9
NDUFB6 NADH-ubiquinone oxydoreductase 1 beta subcomplex 6
NDUFS7 NADH-ubiquinone oxydoreductase FeS protein 7
nm nannometre
NSCLC Non-small-cell-lung carcinoma
OD Optical density
OXPHOS Oxidative phosphorylation
PAGE Polyacrylamide gel electrophoresis
PAR Poly(ADP ribose)
IIIAbbreviations continued
PARP Poly(ADP ribose) polymerase
PBS Phosphate-buffered saline
PFA Paraformaldehyde
PTP Permeability transition pore
RISC RNA-induced silencing complex
RNA Ribonucleic acid
ROS Reactive oxygen species
S.cerevisiae Saccharomyces cerevisiae
SDS Sodium dodecyl sulfate
siRNA small interfering RNA
Smac Second mitochondria-derived activator of caspases
SOD Superoxide dismutase
SSB protein single-strand DNA binding protein
ssDNA single-stranded DNA
ssRNA single-stranded RNA
STS Staurosporine
TAE Tris-acetate EDTA
tBid Truncated Bid
TEM Transmission electron microscopy
TNF Tumor necrosis factor
TOM40 Translocase of outer membrane 40
TRADD TNF-receptor-associated death domain protein
tRNA Transfer RNA
U Units
UV ultra violet
V Volt
VDAC Voltage-dependent anion channel
w/v M
WAH-1 Worm AIF homologue 1
Z-VAD.fmk Benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethylketone
IVContents
1. Introduction ........................................................................................................ 1
1.1. Mitochondria ............................................................................................................ 1
1.1.1. Structure .................................................................................................................................... 2
1.1.2. Multifunctional organelle ........................................................................................................... 2
1.1.3. The respiratory chain................................................................................................................. 3
1.1.3.1. The electron transport ........................................................................................................ 4
1.1.3.2. Structure of the mammalian complex I .............................................................................. 4
1.1.3.3. Assembly of complex I........................................................................................................ 6
1.1.3.4. Mitochondrial diseases....................................................................................................... 6
1.2. Apoptosis................................................................................................................. 7
1.2.1. Signalling pathways leading to apoptosis ................................................................................ 8
1.2.2. Mitochondria and apoptosis.................................................................................................... 10
1.2.2.1. Mitochondrial outer membrane permeabilisation (MOMP) ............................................. 10
1.2.2.2. How do intracellular signals provoke MOMP?................................................................. 11
1.2.2.3. Apoptotic factors released from mitochondria................................................................. 12
1.2.3. Nuclear events during apoptosis ............................................................................................ 13
1.2.4. Caspase-independent apoptosis ............................................................................................ 14
1.2.5. Apoptosis and disease............................................................................................................ 14
1.3. The apoptosis-inducing factor AIF........................................................................ 15
1.3.1. AIF expression, structure, and localisation ............................................................................ 15
1.3.2. Isoforms ................................................................................................................................... 17
1.3.3. Non mammalian orthologues and mammalian homologues................................................. 18
1.3.4. The apoptotic function............................................................................................................. 19
1.3.4.1. Cell-free systems .............................................................................................................. 19
1.3.4.2. In vivo ................................................................................................................................ 20
1.3.4.3. Downregulation of AIF...................................................................................................... 20
1.3.5. AIF translocation...................................................................................................................... 22
1.3.5.1. How is AIF released? ....................................................................................................... 22
1.3.5.2. Are caspases required?.................................................................................................... 24
1.3.5.3. When is AIF released? ..................................................................................................... 25
1.3.6. Control of AIF activity by HSP70 ............................................................................................ 26
1.3.7. How does AIF exert its effect on DNA?.................................................................................. 27
1.3.8. The vital functions.................................................................................................................... 28
1.3.8.1. Radical scavenger function .............................................................................................. 28
1.3.8.2. AIF and cytoplasmic stress granules............................................................................... 29
1.3.9. AIF is a bifunctional protein......................................