Approaches to Detecting False Positives in Yeast Two-Hybrid Systems. (February 2000)

• Record Type: Journal Article
• Authors: Serebriiskii, Ilya; Estojak, Joanne; Berman, Michelle; Golemis, Erica A.
• Is Part Of: Biotechniques. Volume 28:Number 2(2000)
• Page Start: 328
• Page End: 336
• Publication Date: 2000-02
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/00282rr03
• Languages: English
• ISSNs: 0736-6205

Abstract

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 overexpressing 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.