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A B V N M Figure 4: Electron microscopy of Pichia pastoris X33 wild type grown on glycerol (A). Under these conditions mitochondria are well expressed. B: isolated mitochondria from Pichia pastoris grown on glycerol. M, mitochondria; V, vacuole; N, nucleus; Bars. 0.5 and 2 µm. Pictures shown be curtsey of E. Ingolic, FELMI, TU Graz, Austria Doctoral Thesis completed Sabine Rosenberger Phosphatidylethanolamine (PtdEtn) is an essential component in the yeast Saccharomyces cerevisiae required for function and integrity of mitochondrial membranes, for protein import/assembly into the inner mitochondrial membrane, for GPI-anchor synthesis and vacuolar targeting. Biosynthesis of PtdEtn can be accomplished by three different pathways, with the major route mediated by phosphatidylserine decarboxylase 1 (Psd1p) residing in the inner mitochondrial membrane (IMM). Deletion of PSD1 leads to a significant decrease of cellular and mitochondrial PtdEtn thereby conferring a petite phenotype, which is characterized by the loss of respiratory capacity and mitochondrial DNA. To identify a link between the petite phenotype and disturbed lipid homeostasis in mitochondria a petite yeast strain collection was screened for mutants with abnormal phospholipid patterns. This approach unveiled some candidate genes, among them MRPL22. Deletion of MRPL22 leads to a decreased amount of cellular and mitochondrial PtdEtn similar to a psd1 deletion strain. Another major part of this thesis was focused on the assembly of PtdEtn into peroxisomal membranes. To define the traffic routes of PtdEtn from their site of synthesis to peroxisomes single and multiple mutants bearing defects in the different pathways of PtdEtn synthesis were used. PtdEtn formed through all three pathways in different subcellular membranes can be supplied to peroxisomes with comparable efficiency. International Cooperations J. Cregg, Keck Graduate Institute of Applied Life Sciences, Claremont, CA, USA I. Hapala, Slovak Academy of Sciences, Institute of Animal Biochemistry and Genetics, Ivanka pri Dunaji, Slovak Republic V. Tomekova, Department of Medical Chemistry, Biochemistry and Clinical Biochemistry, Faculty of Medicine UPJS, Košice, Slovak Republic 20
T. Hoefken, Institute of Biochemistry, University of Kiel, Germany D. Warnecke, Institute of Botany, University of Hamburg, Germany I. Feussner, Plant Biochemistry, Albrecht-von-Haller-Institute of Plant Sciences, Georg- August University Göttingen, Germany R. Erdmann, Institute of Physiological Chemistry, Ruhr-University Bochum, Germany Research Projects FWF 17321: Phosphatidylethanolamine of yeast mitochondria FWF 18857: Neutral lipid storage and mobilization in the yeast Saccharomyces cerevisiae FWF L-178 (Translational research): Characterization of organelles from the yeast Pichia pastoris FWF PhD Program: Molecular Enzymology Invited Lectures 1) G. Daum Phosphatidylethanolamine, a key lipid of the yeast Comenius University, Bratislava, Slovak Republic, 19 th April 2007 2) G. Daum Neutral lipid storage and mobilization in the yeast Technical University, Bratislava, Slovak Republic, 20 th April 2007 3) G. Daum, T. Czabany, A. Wagner, M. Spanova, S. Rajakumari, K. Grillitsch and K. Athenstaedt Playing the yeast lipid game 8 th Yeast Lipid Conference, Torino, Italy, 10-12 May 2007 4) G. Daum, K. Athenstaedt, A. Wagner, T. Czabany, K. Grillitsch, S. Rajakumari, M. Spanova, D. Zweytick and E. Ingolic Formation and mobilization of neutral lipid depots in the yeast 3 rd European Federation of Biotechnology Conference on Physiology of Yeast and Filamentous Fungi, Helsinki, Finland, 13-16 June 2007 5) G. Daum The Life Cycle of Neutral Lipids: Synthesis, Storage and Degradation Gordon Research Conference “Molecular & Cellular Biology of Lipids“, Waterville Valley Resort, NH, USA, 22-27 July Publications 1) Daum, G., Wagner, A., Czabany, T. and Athenstaedt, K. Dynamics of neutral lipid storage and mobilization in the yeast Biochimie 89 (2007) 243-248 21
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