Using Taxol-sensitized budding yeast to investigate the effect of microtubule stabilization on anaphase onset

Angela R Bunning; Samuel J Anderson; Mohan L Gupta Jr

doi:10.1016/j.xpro.2023.102522

Using Taxol-sensitized budding yeast to investigate the effect of microtubule stabilization on anaphase onset

STAR Protoc. 2023 Sep 15;4(3):102522. doi: 10.1016/j.xpro.2023.102522. Epub 2023 Aug 18.

Authors

Angela R Bunning¹, Samuel J Anderson¹, Mohan L Gupta Jr²

Affiliations

¹ Department of Genetics, Development, and Cell Biology, Iowa State University, Ames, IA 50010, USA.
² Department of Genetics, Development, and Cell Biology, Iowa State University, Ames, IA 50010, USA. Electronic address: mgupta@iastate.edu.

Abstract

The microtubule (MT)-stabilizing drug Taxol (paclitaxel) is a commonly used tool to investigate MT dynamics and MT-dependent processes. Here, we present a protocol for using Taxol-sensitized budding yeast to investigate the effect of microtubule stabilization on anaphase onset. We describe steps for establishing a log phase culture, synchronizing cells in G1, arresting in metaphase, and releasing cells into Taxol. We then detail procedures for imaging and scoring anaphase onset. This protocol facilitates maintenance and reproducibility in testing drug-sensitized and Taxol-sensitized yeast strains. For complete details on the use and execution of this protocol, please refer to Proudfoot et al.¹.

Keywords: Cell Biology; Cell-based Assays; Genetics; Model Organisms.

MeSH terms

Anaphase
Microtubules
Paclitaxel* / pharmacology
Reproducibility of Results
Saccharomyces cerevisiae
Saccharomycetales*

Substances

Paclitaxel