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Fission Yeast as a Platform for Antibacterial Drug Screens Targeting Bacterial Cytoskeleton Proteins

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Published on: April 26, 2024

A yeast strain biosensor to detect cell wall-perturbing agents.

Jose M Rodriguez-Peña¹, Sonia Diez-Muñiz, César Nombela

¹Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense de Madrid, 28040 Madrid, Spain. josemanu@farm.ucm.es

Journal of Biotechnology

|December 7, 2007

Summary

Researchers developed a novel biosensor using the MLP1 gene in yeast to detect cell wall damage. This tool aids in high-throughput screening for new antifungal drugs targeting fungal cell walls.

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Assay for Adhesion and Agar Invasion in S. cerevisiae

Assay for Adhesion and Agar Invasion in S. cerevisiae

Published on: November 8, 2006

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Fission Yeast as a Platform for Antibacterial Drug Screens Targeting Bacterial Cytoskeleton Proteins

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Assay for Adhesion and Agar Invasion in S. cerevisiae

Assay for Adhesion and Agar Invasion in S. cerevisiae

Published on: November 8, 2006

Area of Science:

Mycology
Molecular Biology
Biochemistry

Background:

The fungal cell wall is crucial for fungal survival and a prime target for antifungal therapies.
The Pkc1-mediated cell integrity pathway in Saccharomyces cerevisiae regulates cell wall maintenance.
MLP1 gene expression increases significantly under cell wall stress, serving as an indicator.

Purpose of the Study:

To develop a reporter system for monitoring fungal cell wall integrity.
To create a biosensor for high-throughput screening of antifungal compounds targeting the cell wall.

Main Methods:

Constructed a reporter system using the nourseothricin resistance gene under MLP1 regulatory sequences.
Developed a genetically modified Saccharomyces cerevisiae strain (AT-1) with the reporter integrated into the MLP1 locus.
Tested the AT-1 strain's response to various chemical agents causing cell wall stress.

Main Results:

Yeast cells with the reporter construct showed increased nourseothricin resistance upon cell wall stress due to elevated MLP1 expression.
The AT-1 strain specifically responded to cell wall-perturbing agents, distinguishing them from other growth inhibitors.
The AT-1 strain demonstrated high nourseothricin resistance under stress conditions.

Conclusions:

The developed reporter system and AT-1 strain are effective biosensors for detecting cell wall perturbations in yeast.
This AT-1 strain facilitates high-throughput antifungal drug screening by identifying compounds that specifically target the fungal cell wall.