Nuclear Receptor Research

Investigation of Interactions between DNA and Nuclear Receptors: A Review of the Most Used Methods

Author(s): Juliana Fattori, Nathalia de Carvalho Indolfo, Jéssica Christina Lóis de Oliveira Campos, Natália Bernardi Videira, Aline Villanova Bridi, Tábata Renée Doratioto, Michelle Alexandrino de Assis, and Ana Carolina Migliorini Figueira

Nuclear receptors (NRs) comprise a superfamily of proteins modulated by ligands that regulate the expression of target genes. These proteins share a multidomain structure harboring an N-terminal domain, a highly conserved DNA binding domain, and a ligand binding domain, which has ligand dependent activation function. They play key roles in development, metabolism, and physiology being closely related to diseases. Most of the knowledge about this superfamily emerges from investigations on new ligands and are mostly centered in the ligand binding domain. However, more investigation focusing on interactions between DNA and DNA binding domain is necessary to shed light on important roles of NRs' participation in transcriptional mechanisms and in specific genes network. Here, our goal is to discuss some nuances of NRs-DNA interaction, describing details of the most used techniques in this sort of study, such as gel shift (EMSA), DNA footprinting, reporter gene assay, ChIP-Seq, 3C, and fluorescence anisotropy. Additionally, we aim to provide tools, presenting advantages and disadvantages of these common methods, when choosing the most suitable one to study NRs-DNA interactions to answer specific questions.