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Investigating the function of the microsporidian mitosome using genomics and molecular cell biology


Microsporidia like Trachipleistophora hominis and Encephalitozoon cuniculi, are opportunistic parasites of humans with HIV. Around 50% of cases of serious diarrhoea in AIDS patients are associated with microsporidia - 14 different microsporidia have now been isolated. Despite their importance, the genome and cell biology of microsporidia are poorly understood. I will use a multidisciplinary approach incorporating training in bioinformatics, proteomics, cell and molecular biology to investigate the structu re and function of the recently discovered microsporidian mitosome ' the most highly reduced mitochondrion yet discovered. My work will also further basic understanding of the minimal and essential functions of the mitochondrion itself; an organelle that is vital for eukaryotes. My hypothesis is that the mitosome makes iron sulphur (Fe-S) clusters ' a fundamental process for all eukaryotic cells, and whose dysfunction causes inherited mitochondrial diseases like Friedrichs Ataxia. The E. cuniculi genome en codes seven of the proteins involved in this process, but there is no experimental data on their location or function. I will clone and sequence two of the key genes (Nfs1p, Isu1p) from Trachipleistophora hominis, a more tractable experimental system than Encephalitozoon, and produce antibodies to locate their site of activity. I will also perform in vitro tests of enzyme function. I will do this at Newcastle University with Professor Martin Embley, in whose lab the mitosome was discovered, and where there are state-of-the-art facilities for bioinformatics, cell biology, proteomics and light and electron microscopy. I will also use bioinformatics to search the Encephalitozoon genome for the mitosomal import pathway. Preliminary data suggests that the mitosome may have, uniquely, dispensed with one of the main mitochondrial import pathways. Thus this work will improve understanding of yet another fundamental mitochondrial process ' that of protein import.

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6, Kensington Terrace
United Kingdom

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