Very little is known about the protein composition of protist's non-actin and non-tubulin cytoskeleton, i.e., the different types of intermediate and striated fibres. Yet, this knowledge could solve interesting problems. For example, homologize cytoskeletal structures across distant groups to answer the question of what the ancestral eukaryotic cell looked like or reveal the movement mechanism of the Euglena subsurface corset – the pellicle. The methodology has matured to approach these.
The project aims to prepare cytoskeletal fractions of selected species and analyse their compositions by proteomic means. At the same time, candidate proteins selected from genomic and proteomic data will be localised using specific antibodies and fluorescence expansion microscopy. This allows imaging of the cytoskeleton structures and protein localization with unprecedented resolution. Initial results show that intermediate fibre protein SF-assemblin associates with certain types of microtubular roots in Paratrimastix pyriformis. The structure and composition of Euglena pellicle is partially known from electron microscopy and proteomic analyses and it contains a number of uncharacterised structural or mobile proteins. The project aims to get to the bottom of the mechanism of pellicle movement using in situ labelling and localisation, knock-outs (Crispr-Cas9) and knock-downs (RNAi) of selected protein candidates.
Five relevant publications of the research group:
Zítek J, Füssy Z, Treitli SC, Peña-Diaz P, Vaitová Z, Zavadska D, Harant K, Hampl V. Reduced mitochondria provide an essential function for the cytosolic methionine cycle. Curr Biol. 2022 Nov 2:S0960-9822(22)01673-6.
Soukal P, Hrdá Š, Karnkowska A, Milanowski R, Szabová J, Hradilová M, Strnad H, Vlček Č, Čepička I, Hampl V. Heterotrophic euglenid Rhabdomonas costata resembles its phototrophic relatives in many aspects of molecular and cell biology. Sci Rep. 2021 Jun 22;11(1):13070.
Treitli SC, Peña-Diaz P, Hałakuc P, Karnkowska A, Hampl V. High quality genome assembly of the amitochondriate eukaryote Monocercomonoides exilis. Microb Genom. 2021 Dec;7(12):000745.
Tomečková L, Tomčala A, Oborník M, Hampl V. The Lipid Composition of Euglena gracilis Middle Plastid Membrane Resembles That of Primary Plastid Envelopes. Plant Physiol. 2020 Dec;184(4):2052-2063.
Novák Vanclová AMG, Zoltner M, Kelly S, Soukal P, Záhonová K, Füssy Z, Ebenezer TE, Lacová Dobáková E, Eliáš M, Lukeš J, Field MC, Hampl V. Metabolic quirks and the colourful history of the Euglena gracilis secondary plastid. New Phytol. 2020 Feb;225(4):1578-1592.Apply to the project