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Étude de l'expression des gènes nucléaires codant pour les sous-unités du complexe I mitochondrial humain

Abstract : NADH:ubiquinone oxidoreductase (complex I) is the largest enzymatic complex of the mitochondrial oxidative phosphorylation system (43 human subunits). Few data are available concerning the regulation mechanisms controlling the expression of these proteins.
This thesis started with the study of the promoters of two human complex I genes. The results show that the transcription of NDUFS8 gene encoding the 23 kDa (TYKY) subunit is under the control of the YY1 and Sp1 transcription factors, while the NDUFS7 gene encoding the 20 kDa (PSST) subunit is regulated by NRF-1 and Sp1.
The second part of this study was oriented towards the development of an analytical method of the human mitochondrial proteome by two-dimensional electrophoresis. The aim is to approach the expression of the complex I proteins globally and without a priori: which subunits are regulated in response to an extracellular signal and how ?
Reference human proteome maps were built using mitochondria isolated from placenta in a system involving 2 pH gradients : one for acidic and neutral proteins, one for basic proteins. On
those maps, 85 mitochondrial proteins have been identified using mass spectrometry techniques. Among them, 17 proteins are complex I subunits. Proteomic analysis was then used to study regulation of mitochondrial proteins expression by iron. On a technical point of view, promising results were obtained: high quality 2D-gels were prepared using mitochondria isolated from culture cells and variations in the expression level of complex I and others mitochondrial proteins have been detected. On a biological point of view, preliminary data were obtained. New experiments have toand to analyze the kinetics of the detected variations.
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Submitted on : Wednesday, September 26, 2007 - 4:06:11 PM
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Pierre Lescuyer. Étude de l'expression des gènes nucléaires codant pour les sous-unités du complexe I mitochondrial humain. Sciences du Vivant [q-bio]. Université Joseph-Fourier - Grenoble I, 2002. Français. ⟨tel-00175098⟩

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