Analysis of the pheromone signaling pathway by RT-qPCR in the budding yeast Saccharomyces cerevisiae

Lucía Ramos-Alonso; Ignacio Garcia; Jorrit M Enserink; Pierre Chymkowitch

doi:10.1016/j.xpro.2022.101210

Analysis of the pheromone signaling pathway by RT-qPCR in the budding yeast Saccharomyces cerevisiae

STAR Protoc. 2022 Mar 3;3(1):101210. doi: 10.1016/j.xpro.2022.101210. eCollection 2022 Mar 18.

Authors

Lucía Ramos-Alonso¹, Ignacio Garcia^{2

3}, Jorrit M Enserink^{1

2

3}, Pierre Chymkowitch¹

Affiliations

¹ Section for Biochemistry and Molecular Biology, Faculty of Mathematics and Natural Sciences, University of Oslo, 0316 Oslo, Norway.
² Department of Molecular Cell Biology, Institute for Cancer Research, Oslo University Hospital, 0379 Oslo, Norway.
³ Centre for Cancer Cell Reprogramming, Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, 0318 Oslo, Norway.

Abstract

FUS3 and STE2 expression levels can be used as reporters for signaling through the pheromone pathway in the budding yeast Saccharomyces cerevisiae. Here, we describe an optimized protocol to measure the expression levels of FUS3 and STE2 using quantitative reverse transcription PCR (RT-qPCR). We describe the steps for comparing untreated and pheromone-treated yeast cells and how to quantify the changes in various deletion strains. The protocol can be applied to determine potential regulators of the pheromone pathway. For complete details on the use and execution of this protocol, please refer to Garcia et al. (2021).

Keywords: Cell Biology; Gene Expression; Model Organisms; Molecular Biology; Signal Transduction.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Mitogen-Activated Protein Kinases / metabolism
Pheromones / metabolism
Saccharomyces cerevisiae / genetics
Saccharomyces cerevisiae Proteins* / genetics
Signal Transduction / genetics
Yeast, Dried*

Substances

Pheromones
Saccharomyces cerevisiae Proteins
FUS3 protein, S cerevisiae
Mitogen-Activated Protein Kinases