Structure and function of proteins participating in the homeostasis of alkali metal cations and intracellular pH
Ion homeostasis is a crucial requirement for all cells. They have developed efficient transport systems to acquire and store the desired elements and robust mechanisms to maintain homeostatic concentrations to avoid toxicity. Tightly regulated alkali-metal-cation and proton fluxes across the plasma and organellar membranes are also indispensable for the maintenance of proper membrane potential and intracellular pH.
The aim of this PhD thesis will be the identification and characterization of so far unknown proteins that participate in biogenesis, degradation and activity regulation of transporters involved in alkali-metal-cation and pH homeostasis, using yeast cells as a model and a host. The work will consist of library screens, construction of knock-out strains, expression vectors, and the characterization of their phenotype and physiological parameters.
Candidate’s profile (requirements):
The candidate should be fluent in English, highly self-motivated with master's degree or equivalent (obtained before October 2018) in molecular biology, biochemistry, microbiology, pharmacology or related fields. Experience in basic laboratory (PCR, DNA electrophoresis, bacteria transformation, plasmid isolation) and bioinformatics (DNA and protein sequence search and comparison, sequenced fragments analysis, plasmid and primers design) techniques is an advantage.
Arino J., Ramos J., Sychrova H. Alkali-metal-cation transport and homeostasis in yeasts. Microbiol. Mol. Biol. Rev. 75 (1), 95-120 (2010).
Kinclova-Zimmermannová O., Falson P., Cmunt D., Sychrova H. A hydrophobic filter confers the cation selectivity of Zygosaccharomyces rouxii plasma-membrane Na+/H+ antiporters. J. Mol. Biol. 427: 1681-1694 (2015)
Kodedová M., Sychrová H. Changes in the sterol composition of the plasma membrane affect membrane potential, salt tolerance and the activity of multidrug resistance pumps in Saccharomyces cerevisiae. PLoS ONE 10(9), e0139306 (2015).
Llopis-Torregrosa V., Hušeková B., Sychrová H. Potassium uptake mediated by Trk1 is crucial for Candida glabrata growth and fitness PLoS ONE 11(4): e0153374 (2016).
Felcmanova K., Neveceralova P., Sychrova H., ZimmermannovaO. Yeast Kch1 and Kch2 membrane proteins play a pleiotropic role in membrane potential establishment and monovalent cation homeostasis regulation. FEMS Yeast Res, 17: fox053 (2017).
Supervisor: Hana Sychrová, Ph.D., D.Sc.