Utilizing chemically induced dimerization of FKBP to analyze endocytosis by live-cell imaging in budding yeast

Chemically induced dimerization (CID) is a useful tool for artificially inducing protein-protein interactions. Although CID has been used extensively for live-cell microscopy applications in mammalian systems, it is rarely utilized in yeast cell biology studies. Here, we present a step-by-step proto...

Full description

Saved in:
Bibliographic Details
Published in:STAR protocols 2022-06, Vol.3 (2), p.101323-101323, Article 101323
Main Authors: Lamb, Andrew K., Di Pietro, Santiago M.
Format: Article
Language:English
Subjects:
Animals
Cell Biology
Cell-based Assays
Dimerization
Endocytosis
Mammals
Microscopy
Model Organisms
Protocol
Saccharomyces cerevisiae
Saccharomycetales
Sheep
Signal Transduction
Tacrolimus Binding Proteins
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Chemically induced dimerization (CID) is a useful tool for artificially inducing protein-protein interactions. Although CID has been used extensively for live-cell microscopy applications in mammalian systems, it is rarely utilized in yeast cell biology studies. Here, we present a step-by-step protocol for the utilization of a CID system in live-cell microscopy experiments of budding yeast endocytosis. While focusing on the study of endocytosis, this protocol framework is adaptable to the study of other cellular processes in Saccharomyces cerevisiae. For complete details on the use and execution of this protocol, please refer to Lamb et al. (2021). [Display omitted] •Generation of yeast strains for endogenous expression of FKBP-tagged proteins•Utilization of an inducible homodimerization system in S. cerevisiae•Fluorescence microscopy imaging of clathrin-mediated endocytosis Publisher's note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Chemically induced dimerization (CID) is a useful tool for artificially inducing protein-protein interactions. Although CID has been used extensively for live-cell microscopy applications in mammalian systems, it is rarely utilized in yeast cell biology studies. Here, we present a step-by-step protocol for the utilization of a CID system in live-cell microscopy experiments of budding yeast endocytosis. While focusing on the study of endocytosis, this protocol framework is adaptable to the study of other cellular processes in Saccharomyces cerevisiae.
ISSN:2666-1667
2666-1667
DOI:10.1016/j.xpro.2022.101323