Use of different microscopy techniques for the study of stress response in exomer mutants in Schizosaccharomyces pombe

Autor: Moscoso-Romero, Esteban, Duque, Alicia, Valdivieso, María Henar
Rok vydání: 2022
Popis: Trabajo presentado en el Spanish and Portuguese Advanced Optical Microscopy Meeting (SPAOM), celebrado en Salamanca (España), del 23 al 25 de noviembre de 2022
Plasma membrane (PM) and membranous organelles contribute to the physiology of the Eukaryotic cell by participating in vesicle trafficking and the maintenance of ion homeostasis. Exomer is a protein complex that was described in Saccharomyces cerevisiae as essential for the transport on transmembrane proteins from the trans-Golgi network to the plasma membrane, such that its absence leads to the retention of a set of selected cargoes in this organelle. However, this retention does not explain all phenotypes observed in exomer mutants. In this work, we use of several microscopy techniques to understand the role of exomer in stress response in Schizosaccharomyces pombe. Thus, we have compared the results obtained with a conventional fluorescence microscope to those obtained with a confocal Spinning disk microscope. Also, we have used slides and chambers of different shape and material (plastic, glass) to mount and visualize the samples. Using these techniques we have found three different phenotypes in exomer mutants treated with 0.6 M KCl: i) a defect in the PM association of several regulators of the cell integrity pathway (CIP), a Mitogen-Activated Protein Kinase (MAPK) pathway. ii) a defective localization of some ion transporters, like the K+ exporting ATPase Cta3. These transporters are polarized to the cell tips and equator; however, their distribution is asymmetrical ad/or subapical (Figure 1). iii) a defect in the distribution and of some (PM) lipids, resulting in altered lipid environments for the PM integral proteins. [1] T. Nishikawa, H. Aiba, and T. Mizuno. The cta3+ gene that encodes a cation-transporting P-type ATPase is induced by salt stress under control of the Wis1-Sty1 MAPKK-MAPK cascade in fission yeast. FEBS Letters. 455, 183-187 (1999). [2] R. Sugiura et al. Functional link between Rab GTPase-mediated membrane trafficking and PI4,5P2 signaling. Genes to cells. 19, 177-197 (2014).
Databáze: OpenAIRE
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