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7e colloque du réseau MeetOchondrie 4 – 7 mai 2014 – Evian Table Ronde Dynamique Mitochondriale
Manuel Rojo Institut de Biochimie et Génétique Cellulaires UMR 5095 CNRS – Université de Bordeaux
Dynamique fonctionnelle : acteurs et pertinence physiologique de la dynamique mitochondriale
matrice!
membrane!interne!
membrane!externe!
espace intermembranaire!Alberts et al. 1983!
Les mitochondries: ultrastructure!
(1999)!
(1985)!
m.rojo /
mouse heart mouse hepatocyte snail epithelial cell
Morphologie, Distribution et Ultrastructure mitochondriales
(Pierre Favard dans Alberts, Molecular Biology of the Cell)
m.rojo /
JCB 2009
- M, Safiulina D, Szabadkai G, Das S, Fransson A, Aspenstrom P, Rizzuto R, Hajnóczky G. (2008) Bidirectional Ca2+-dependent control of mitochondrial dynamics by the Miro GTPase.Saotome . Proc Natl Acad Sci U S A.105(52):20728-33. - MacAskill, A.F., Rinholm, J.E., Twelvetrees, A.E., Arancibia-Carcamo, I.L., Muir, J., Fransson, Å., Aspenström, P., Attwell, D. and Kittler, J. (2009) Miro1 is a calcium sensor for glutamate receptor-dependent localization of mitochondria at synapses. Neuron 61, 541–555. - Wang, X. and Schwarz, T.L. (2009) The mechanism of Ca2+-dependent regulation of kinesin-mediated mitochondrial motility. Cell 136, 163–174.
Miro forms a complex with Milton to recruit kinesin to the mitochondria Miro–Milton complex supports mitochondrial motility along microtubules Miro–Milton complex confers Ca2+ sensitivity to mitochondrial motility
JCB 2006
microtubule
Miro/Gem1p
m.rojo /
JCB 2009
- M, Safiulina D, Szabadkai G, Das S, Fransson A, Aspenstrom P, Rizzuto R, Hajnóczky G. (2008) Bidirectional Ca2+-dependent control of mitochondrial dynamics by the Miro GTPase.Saotome . Proc Natl Acad Sci U S A.105(52):20728-33. - MacAskill, A.F., Rinholm, J.E., Twelvetrees, A.E., Arancibia-Carcamo, I.L., Muir, J., Fransson, Å., Aspenström, P., Attwell, D. and Kittler, J. (2009) Miro1 is a calcium sensor for glutamate receptor-dependent localization of mitochondria at synapses. Neuron 61, 541–555. - Wang, X. and Schwarz, T.L. (2009) The mechanism of Ca2+-dependent regulation of kinesin-mediated mitochondrial motility. Cell 136, 163–174.
Miro forms a complex with Milton to recruit kinesin to the mitochondria Miro–Milton complex supports mitochondrial motility along microtubules Miro–Milton complex confers Ca2+ sensitivity to mitochondrial motility
JCB 2006
microtubule
Miro/Gem1p
m.rojo /
2007
anterograde: Arp2/3+Jsn1p/Puf1p+Puf3p
retrograde: Arp2/3 + typeII Myosin (Myo2)
bud
actin filament
2011
actin filament
m.rojo /
2007
anterograde: Arp2/3+Jsn1p/Puf1p+Puf3p
retrograde: Arp2/3 + typeII Myosin (Myo2)
bud
actin filament
2011
actin filament
m.rojo /
m.rojo /
Drp1, Mff, Fis1, ….
Mfn1, Mfn2, OPA1, ….
* dynamin-related proteins
mitochondrial dynamics <-> a futile cycle of fusion & fission
cell fusion &
mitochondrial fusion
m.rojo /
Drp1, Mff, Fis1, ….
Mfn1, Mfn2, OPA1, ….
mitochondrial fusion: a "#m-dependent process
cell fusion + cccp (no "#m) - cccp
"$m is essential for inner membrane (IM) fusion
"$m is dispensable for outer membrane (OM) fusion
m.rojo /
De-energized mitochondria are excluded from the network of energized, fusogenic mitochondria This allows their selective targeting for degradation.
Drp1 Fis1 Mff
Drp1 Fis1 Mff
Mfn1 Mfn2 OPA1
Mfn1 Mfn2 OPA1
However, for such models to apply, fusion inhibition must be: - dominant - occur with physiological/pathological OXPHOS defects.
Twig et al 2008
yeast S. cerevisiae ! ! !mammals!! ! !!
facultative anaerob ! ! ! !obligate aerobic !!linear mtDNA molecules ! ! !circular mtDNA molecules!~ 80 kb ; 20 - 50 copies / cell ! ! !16.5 kb ; 1000s of copies / cell !4 respiratory complexes (no complex I) ! !5 respiratory complexes!cell ~ 3-6 µm ! ! ! !cell ≥ ~ 30 - 50 µm !!
Fzo1p* ! ! ! ! !Mfn1*, Mfn2* !!Mgm1p* ! ! ! ! !OPA1* ! !!Ugo1p ! ! ! ! !--------!-------! ! ! ! ! !Mito-PLD!-------! ! ! ! ! !Bcl2-family proteins !!
Dnm1p* ! ! ! ! !Drp1/Dlp1*!!Fis1p! ! ! ! ! !Fis1!----- ! ! ! ! ! !Mff!----- ! ! ! ! ! !MiD49 & MiD51 !!Mdv1p/Net2p ! ! ! !------ ! !!Caf4p ! ! ! ! !------!------ ! ! ! ! ! !GDAP1!------ ! ! ! ! ! !MTP18!
Mdm10p/Mdm12p/Mmm1p ! ! !------!Mdm31/Mdm32 ! ! ! !------!Gem1p ! ! ! ! !------!
Mitochondrial fusion/fission machineries
m.rojo / * GTP-binding proteins of the dynamin-superfamily
fusion
fission
“tubulation” or otherwise unknown…
Sesaki H, Jensen RE (1999) Division versus Fusion: Dnm1p and Fzo1p Antagonistically Regulate Mitochondrial Shape JCell Biol 147:699-706
Sesaki H, Southard SM, Yaffe MP, Jensen RE (2003) Mgm1p, a dynamin-related GTPase, is essential for fusion of the mitochondrial outer membrane MolBiolCell 14:2342-256.
WT mgm1"% dnm11"% mgm1"/dnm11"%
Antagonistic activity fusion and fission determine mitochondrial morphology S. cerevisiae
m.rojo /
Wakabayashi et al. (2009). The dynamin-related GTPase Drp1 is required for embryonic and brain development in mice. The Journal of Cell Biology, 186(6), 805–816.
Antagonistic activity fusion and fission determine mitochondrial morphology mammals
DRP1
Koshiba, et al. (2004). Structural basis of mitochondrial tethering by mitofusin complexes. Science (New York, NY), 305(5685), 858–862. doi:10.1126/science.1099793
MFN1 MFN2
wt DKO
m.rojo /
mitochondria!secretory and endocytic organelles!
ER!
Golgi!
PM!
endosomes!
lysosomes!
nucleus!
budding/fission!
fusion!movement!
budding/fission!
maturation!
maturation, budding and cargo selection vectorial vesicular traffic
heterotypic fusion rabs & SNAREs
mitochondria!
peroxisomes!
fusion/fission!
fusion/fission!
?!?!
?!
?!
homotypic fusion & fission no rabs & no SNAREs
m.rojo /
Interaction(s) of mitochondria with other organelles?
2009
Mmm1, Mdm10, Mdm12 + Gem1p - “tubulation” mutants of mitochondrial distribution and morphology (mdm) (1994) - association with actin cables (2003) - mtDNA-nucleoid maintenance (2004) - assembly of OM proteins (2004 – 2007)
ERMES: ER-mitochondria Encounter Structure -> non-vesicular lipid exchange -> Ca2+ exchange
2011
ER-mitochondrial connections/interactions (S.cerevisiae)
m.rojo /
What is the physio(patho)logical relevance of mitochondrial dynamics? (animal models: KO and mutations)
m.rojo /
Sauvanet et al. (2010) Dynamique et morphologie mitochondriales. Acteurs, mécanismes et pertinence fonctionnelle Med Sci (Paris). 26(10):823-9.
KO Fzo1/Mfn: severe physiological and developmental consequences
KO Mgm1/OPA1: severe physiological and developmental consequences
KO Dnm1/DRP1: severe physiological and developmental consequences
What is the physio(patho)logical relevance of mitochondrial dynamics? (animal models: KO and mutations)
m.rojo /
Sauvanet et al. (2010) Dynamique et morphologie mitochondriales. Acteurs, mécanismes et pertinence fonctionnelle Med Sci (Paris). 26(10):823-9.
KO Fzo1/Mfn: severe physiological and developmental consequences
KO Mgm1/OPA1: severe physiological and developmental consequences
KO Dnm1/DRP1: severe physiological and developmental consequences
m.rojo /
mitochondrial distribution, morphology & dynamics 2014
autophagy-mitophagy
mitochondrial DNA transmission & segregation
cellular & mitochondrial Bioenergetics
OXPHOS-diseases
proliferation apoptosis
Ca2+-signalling ROS production & consumption
senescence
signaling regulation
innate immunity
neuronal function - neuropathies morphogenesis of spines and synapses
Lymphocyte chemotaxis
Alzheimer Huntington Parkinson
obesity diabetes Insulin release
viral infection
Cardiomyocite development
function of pancreatic beta-cells Insulin secretion
m.rojo /
mitochondrial distribution, morphology & dynamics 2014
cellular & mitochondrial Bioenergetics
OXPHOS-diseases
proliferation apoptosis
neuronal function - neuropathies morphogenesis of spines and
synapses
Alzheimer Huntington Parkinson
Mitochondrial dynamics and hereditary neuro(muscular) diseases
m.rojo /
MFN2: maladie de Charcot-Marie-Tooth de type 2A (CMT2A)
OPA1: maladie de Kjer, atrophie autosomique dominante de type 1 (DOA)
DRP1: syndrome néonatal létal
GDAP1: maladie de Charcot-Marie-Tooth de type 4A et 2K (CMT4A et CMT2K)
Mitochondrial dynamics and hereditary neuro(muscular) diseases
m.rojo /
MFN2: maladie de Charcot-Marie-Tooth de type 2A (CMT2A)
OPA1: maladie de Kjer, atrophie autosomique dominante de type 1 (DOA)
DRP1: syndrome néonatal létal
GDAP1: maladie de Charcot-Marie-Tooth de type 4A et 2K (CMT4A et CMT2K)
Lenaers et al. (2010). De la levure aux maladies neurodégénératives. Dix ans d’exploration des pathologies de la dynamique mitochondriale. Médecine Sciences : M/S, 26(10), 836–841.
m.rojo /
mitochondrial distribution, morphology & dynamics 2014
cellular & mitochondrial Bioenergetics
OXPHOS-diseases
proliferation apoptosis
neuronal function - neuropathies morphogenesis of spines and
synapses
Alzheimer Huntington Parkinson
Mitochondrial fusion / fission and bioenergetics
m.rojo /
This inhibition enables exclusion of defective mitochondria and their selective targeting to mitophagy.
2012
2008
m.rojo /
mitochondrial distribution, morphology & dynamics 2014
cellular & mitochondrial Bioenergetics
OXPHOS-diseases
proliferation apoptosis
neuronal function - neuropathies morphogenesis of spines and
synapses
Alzheimer Huntington Parkinson
m.rojo /
mitochondrial distribution, morphology & dynamics 2014
cellular & mitochondrial Bioenergetics
OXPHOS-diseases
proliferation apoptosis
neuronal function - neuropathies morphogenesis of spines and
synapses
Alzheimer Huntington Parkinson
m.rojo /
Voire la dynamique mitochondriale
autophagy-mitophagy
mitochondrial DNA transmission & segregation
cellular & mitochondrial Bioenergetics
OXPHOS-diseases
proliferation apoptosis
Ca2+-signalling ROS production & consumption
senescence
signaling regulation
innate immunity
neuronal function - neuropathies morphogenesis of spines and synapses
Lymphocyte chemotaxis
Alzheimer Huntington Parkinson
obesity diabetes Insulin release
viral infection
Cardiomyocite development
function of pancreatic beta-cells Insulin secretion
m.rojo /
Lenaers et al. (2010). De la levure aux maladies neurodégénératives. Dix ans d’exploration des pathologies de la dynamique mitochondriale. Médecine Sciences : M/S, 26(10), 836–841.
Agier et al. (2012). Defective mitochondrial fusion, altered respiratory function, and distorted cristae structure in skin fibroblasts with heterozygous OPA1 mutations. Biochimica Et Biophysica Acta, 1822(10), 1570–1580.
Voire la morphologie mitochondriale – mutations d’OPA1 - mitochondrial morphology varies between alleles -
m.rojo /
Mishra et al. (2014). Proteolytic cleavage of opa1 stimulates mitochondrial inner membrane fusion and couples fusion to oxidative phosphorylation. Cell Metabolism, 19(4), 630–641
Guillery et al. (2008). Modulation of mitochondrial morphology by bioenergetics defects in primary human fibroblasts. Neuromuscular Disorders : NMD, 18(4), 319–330.
WT
ND1
glucose deoxy-glucose
Voire la morphologie mitochondriale – deficits de l’OXPHOS - specific morphology requires specific culture conditions -
IBGC – Bordeaux Jean-Paul di Rago
Stéphane Duvezin-Caubet
Patrick Paumard!Marie-France Giraud!
Stéphen Manon Muriel Priault
Bénédicte Salin, Corinne Blancard!
IBGC – Bordeaux Organisation et Dynamique Mitochondriales
Marine Bouhier Claudine David
Manuel Rojo
Cécile Sauvanet (2008 – 2011)
Institut Jacques Monod David Tareste
notre équipe collaborations