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Approaches to detecting false positives in yeast two-hybrid systems
I Serebriiskii1, J Estojak, M Berman
1Fox Chase Cancer Center, Philadelphia, PA, USA.
Biotechniques
|February 23, 2000
Summary
False positives in yeast two-hybrid screens can arise from indirect metabolic effects, not true protein interactions. This study identifies strategies to distinguish these false positives, improving the reliability of protein interaction data.
Area of Science:
- Molecular Biology
- Yeast Genetics
- Protein Interaction Studies
Background:
- Two-hybrid screens are widely used to identify protein-protein interactions.
- However, some identified interactions may not be biologically relevant, leading to false positives.
- The underlying causes for these false positives are often unclear.
Purpose of the Study:
- To investigate the reasons behind false positive results in yeast two-hybrid screens.
- To determine if indirect metabolic effects contribute to false positive yeast two-hybrid interactions.
- To develop strategies for identifying and mitigating such false positives.
Main Methods:
- Over-expression of putative false positive clones in yeast.
- Assessment of biological effects in yeast, including growth rate and cell permeability.
- Analysis of reporter gene activity (e.g., LacZ) in response to clone expression.
Main Results:
- Expression of certain clones induced significant biological effects in yeast.
- These effects included altered growth rates and cell permeability.
- Such indirect metabolic effects can bias reporter gene activity, mimicking true interactions.
Conclusions:
- Indirect metabolic effects can cause false positive results in yeast two-hybrid screens.
- Four strategies were identified to help determine if a protein was selected due to metabolic effects.
- These strategies can improve the accuracy of protein interaction data derived from two-hybrid screens.