Rapid hyphal growth of filamentous fungi, including many pathogens of plant crops, animals, and humans, depends on a distinctive dynamic structure near the growing hyphal cell tip, known as the Spitzenkörper (SPK). The SPK, which contains an accumulation of secretory vesicles, ribosomes and other poorly characterised material, serves as a link, or clearing-house, between intracellular transport and supply of vesicles to the secretory machinery at the hyphal tip. While some of the protein components of the SPK are known, to date it has not been analysed using comprehensive proteomics technologies. The PhD project will involve multiple "omics" approaches to characterizing the SPK in filamentous fungi, in order to define its protein composition, starting with non-pathogenic model systems (e.g. A. nidulans) as proof-of-concept, and ideally later continuing on to pathogens. The student will be able to test the role of individual SPK proteins in vivo by reverse-genetics approaches. In addition, there is indirect evidence that the SPK could serve as a factory for the synthesis of a subset of proteins involved in secretion and hyphal growth, via localised translation of a selection of mRNAs. The student will investigate this by analysis and functional characterisation of the SPK-associated transcriptome. A deeper understanding of SPK organisation and function may ultimately lead to improvements in crop production and human/animal health.
Methods involved in the project will include: gene tagging, live-cell fluorescence microscopy, In vivo post-translational labelling, protein purification, mass spectrometry/proteomics, RNAseq, and computer-based analysis.
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Haag, C., Steuten, B., and Feldbrugge, M. (2015). Membrane-Coupled mRNA Trafficking in Fungi. Annu Rev Microbiol 69, 265-281.
Langdon, E.M., and Gladfelter, A.S. (2018). A New Lens for RNA Localization: Liquid-Liquid Phase Separation. Annu Rev Microbiol 72, 255-271.
Riquelme, M., and Sanchez-Leon, E. (2014). The Spitzenkorper: a choreographer of fungal growth and morphogenesis. Curr Opin Microbiol 20, 27-33.
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FTE Category A staff submitted: 109.70
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