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Yeast Tripartite Biosensors Sensitive to Protein Stability and Aggregation Propensity.
Veronika Sachsenhauser1,2, Xiexiong Deng1, Hyun-Hee Kim1
1Department of Molecular, Cellular, and Developmental Biology and Howard Hughes Medical Institute, University of Michigan, Ann Arbor, Michigan 48109-1085, United States.
ACS Chemical Biology
|February 28, 2020
Summary
Researchers developed a novel biosensor to study protein misfolding and aggregation within living yeast cells. This system links protein stability to antibiotic resistance, enabling in vivo analysis of disease-associated proteins.
Area of Science:
- Biochemistry
- Cell Biology
- Molecular Biology
Background:
- Studying protein misfolding and aggregation in vitro is common, but cellular context studies are limited.
- The cellular environment complicates many traditional biophysical techniques.
- A need exists for tools to assess protein stability and aggregation in vivo.
Purpose of the Study:
- To develop a novel biosensor system for assessing in vivo protein stability and solubility.
- To link protein aggregation susceptibility directly to a measurable cellular output.
- To investigate the in vivo behavior of disease-associated proteins in yeast.
Main Methods:
- A tripartite fusion biosensor approach was designed.
- Proteins of interest were inserted into antibiotic resistance markers.
- The system directly correlates protein aggregation with antibiotic resistance readouts.
Main Results:
- A linear relationship was established between thermodynamic stabilities of model protein variants and antibiotic resistance.
- The biosensor successfully assessed in vivo properties of yeast prion proteins (Sup35, Rnq1).
- Aggregation-prone proteins linked to neurodegenerative diseases (Aβ42, α-synuclein) were studied in vivo.
Conclusions:
- The tripartite fusion biosensor is a viable tool for studying in vivo protein stability and aggregation.
- This system provides a new method for investigating protein behavior in the cellular context.
- The approach is applicable to studying yeast prions and proteins implicated in neurodegenerative disorders.
