A quantitative dissection of the DNA-binding properties of pathogenic GATA1 mutants

Abstract

Transcription factors are modulated by a precisely coordinated set of conditions, including cell context, target sequences and their accessibility, and co-factor recruitment. Disruption to any of these conditions can dramatically affect transcription factor activity, but quantitatively characterizing the consequences of individual mutations—either in the transcription factors themselves or in their target sequences—has remained a technical challenge. Zambo et al. present an innovation on their native holdup assay that measures DNA–protein binding activity under physiologic or near-physiologic conditions and use mutant GATA1-ATP2B4 binding as an illustrative example. This technique holds promise for uncovering the molecular mechanisms underlying genetically-driven diseases.