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Project 2

Understanding the limits of mitochondrial reductive evolution and identifying the critical interdependencies between organelle and host cell

PhD position. Supervisors: Martin Embley (UniNewcastle) and Thomas Schweder (UniGreifswald) (with input from Robert Hirt, Alina Goldberg and Shaojun Long, UniNewcastle). Host: UniNewcastle. InternshipSecondments: to UniGreifswald.
Rationale

The minimal mitochondria (mitosomes) of obligate intracellular microsporidian parasitic protists have lost many of the pathways and proteins that make mitochondria essential components of our own cells. Mitosomes are thus excellent model systems for understanding the limits of mitochondrial reductive evolution, and for identifying the truly essential functions of mitochondria.

Objectives

The PhD student will (i) use comparative bioinformatics of microsporidian genomes to discriminate general and lineage-specific features of mitosome reductive evolution, (ii) identify in silico potential metabolic interdependencies between mitosomes and the microsporidian cell, and (iii) test hypotheses of mitosome functions using confocal and electron microscopy, functional assays and proteomics (UniGreifswald).

Key methods

A database of experimentally verified mitochondrial proteins from diverse eukaryotes will be used to generate models for the identification of divergent microsporidian homologues. These analyses will be used to reconstruct in silico putative mitosomal proteomes for each microsporidian, and will be complemented by proteomics analysis of mitosomes (UniGreifswald). The location of candidate mitosomal pathways will be verified by indirect fluorescence immuno-localization or immuno-electron microscopy of key proteins. Protein function will be investigated by functional assays and complementation of yeast mitochondrial mutants. Transport proteins, which form a vital link between organelle and cytosol, will be investigated for their substrate specificities using a variety of transport assays in heterologous hosts and/or liposomes.