Two-hybrid system

This protocol describes a nondestructive FRET-based assay for measuring protein interactions in cultured cells. The correlation between FRET efficiencies and relative protein concentration can be used to determine relative binding affinities.

The centrosome is a large intracellular structure that serves as the microtubule-organising center, but how it is accurately assembled is not known. Here the authors generate a ‘domain-level’ centrosome interactome and show that Plk4 positions the essential centriole component Asterless by phosphorylating Cep135.

A high-sensitivity, high-quality yeast two-hybrid screen using short-read sequencing as its readout is used to map the interactome of human methyltransferases.

An almost-complete, sequence-verified collection of Arabidopsis thaliana root stele transcription factors is reported. The authors use it in the enhanced yeast-one hybrid (eY1H) assay to map gene regulatory interactions in the plant. Also in this issue, Reece-Hoyes et al. describe the eY1H pipeline.

A sequence-verified collection of human transcription factors is reported. The authors used it in the enhanced yeast-one hybrid (eY1H) assay to map human gene regulatory networks. Also in this issue, Reece-Hoyes et al. describe the eY1H pipeline.