Light-dependent inhibition of clathrin-mediated endocytosis in yeast
Prischich, D.; Encinar del Dedo, J.; Cambra, M.; Prat, J.; Camarero, N.; Nevola, L.; Martin-Quiros, A.; Rebollo, E.; Giralt, E.; Geli, M. I.; Gorostiza, P.
Show abstract
Clathrin-mediated endocytosis (CME) is an essential cellular process, which is evolutionarily conserved among eukaryotes. Yeast constitutes a powerful genetic model to dissect the complex endocytic machinery, yet there is a lack of pharmacological agents that could complement genetics in selectively and reversibly interfere with CME in these organisms. TL2 is a light-regulated peptide inhibitor that targets the AP2/{beta}-arrestin interaction and that can photocontrol CME with high spatiotemporal precision in mammalian cells. Here, we study endocytic protein dynamics by live-cell imaging of the fluorescently tagged coat-associated protein Sla1-GFP and demonstrate that TL2 retains its inhibitory activity in S. cerevisiae spheroplasts, thus providing a unique tool for acute and reversible CME modulation in yeast.
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