Intrinsic disorder of Drosophila melanogaster hormone receptor 38 N‐terminal domain

Abstract

Drosophila hormone receptor 38 (dHR38), an ortholog of the vertebrate NR4A subclass of nuclear receptors, responds to ecdysteroids, which mediate developmental transitions during the Drosophila life cycle. However, this response is independent of the ecdysteroid receptor, and it does not involve binding of ecdysteroids to dHR38. It has been suggested that ecdysteroids may indirectly activate dHR38, perhaps by recruiting specific proteins. There have been recent reports pointing out the decisive role that nuclear receptor N-terminal domains (NTDs) have in protein-protein interactions that are important for regulation of gene expression. It is reasonable to assume that dHR38-NTD may also be involved in some protein-protein interactions that are critical for the ecdysteroid signaling pathway. To facilitate the exploration of the molecular basis of these interactions, we developed and optimized a protocol for the efficient expression and purification of the recombinant dHR38-NTD. Using a diverse array of biochemical and biophysical methods, we carried out the first structural characterization of dHR38-NTD. The results of our study indicate that dHR38-NTD exhibits a characteristic reminiscent of pre-molten globule-like intrinsically disordered proteins existing in a partially unfolded conformation with regions of secondary structures. The dHR38-NTD structure, which apparently comprises some local, ordered, tertiary structure clusters, is pliable and can adopt more ordered conformations in response to changes in environmental conditions. Thus, dHR38-NTD, which exhibits the structural and functional characteristic of a pre-molten globule-like intrinsically disordered protein, could serve as a platform for multiple protein-protein interactions, possibly including interactions with proteins involved in an unusual ecdysteroid signaling pathway.