Approaches to detecting false positives in yeast two-hybrid systems.
While many novel associations predicted by two-hybrid library screens reflect actual biological associations of two proteins in vivo, at times the functional co-relevance of two proteins scored as interacting in the two-hybrid system is unlikely. The reason for this positive score remains obscure, which leads to designating such clones as false positives. After investigating the effect of over-expressing a series of putative false positives in yeast, we determined that expression of some of these clones induces an array of biological effects in yeast, including altered growth rate and cell permeability, that bias perceived activity of LacZ reporters. Based on these observations, we identify four simple strategies that can assist in determining whether a protein is likely to have been selected in a two-hybrid screen because of indirect metabolic effects.
Serebriiskii
I
I
Fox Chase Cancer Center, Philadelphia, PA, USA.
Estojak
J
J
Berman
M
M
Golemis
E A
EA
D001709:Biotechnology; D002463:Cell Membrane Permeability; D005189:False Positive Reactions; D007763:Lac Operon; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D015533:Transcriptional Activation; D020798:Two-Hybrid System Techniques / Q000706:statistics & numerical data*