The development of the Heterodimeric enhanced Yeast-One Hybrid (HD-eY1H) assay

Abstract

[EMBARGOED UNTIL 6/1/2023] Life can be summarized as a continuous struggle to maintain homeostasis - or balance - despite dynamic changes in environmental conditions. Changes in gene expression is one of the many ways organisms can interact and respond to their environment to adapt and preserve homeostasis. The central dogma of biology tells us about the flow of genetic information from DNA to RNA and then from RNA to proteins, who ultimately execute metabolic processes. Understanding the mechanisms that dictate gene expression offers the possibility of untangling the complex regulatory networks that have evolved over time. And these regulatory networks cover all aspects of survival for an organism and are what allow life to fleetingly maintain homeostasis. This thesis summarizes the development of the Heterodimeric eYeast-One Hybrid (HD-eY1H) assay, which was developed to identify novel protein-protein-DNA interactions. As a proof of concept, we used HD-eY1H to explore the iron homeostatic network in the reference plant Arabidopsis thaliana.