Corticosteroid Activation of Atlantic Sea Lamprey Corticoid Receptor: Allosteric Regulation by the N-terminal Domain

TL;DR

This study characterizes the activation of the ancestral corticoid receptor in lampreys by corticosteroids, revealing allosteric regulation by the N-terminal domain and its evolutionary relationship to other vertebrate steroid receptors.

Contribution

It identifies the full-length sequences of lamprey CR, explores their steroid specificity, and demonstrates the N-terminal domain's allosteric regulatory role on transcriptional activity.

Findings

CR1 and CR2 respond strongly to 11-deoxycorticosterone and 11-deoxycortisol.

Deletion of NTD reduces activation with MMTV promoter but increases it with TAT3 promoter.

Lamprey CR is evolutionarily close to elephant shark MR, distant from shark GR.

Abstract

Lampreys are jawless fish that evolved about 550 million years ago at the base of the vertebrate line. Modern lampreys contain a corticoid receptor (CR), the common ancestor of the glucocorticoid receptor (GR) and mineralocorticoid receptor (MR), which first appear in cartilaginous fish, such as sharks. Until recently, 344 amino acids at the amino terminus of adult lamprey CR were not present in the lamprey CR sequence in GenBank. A search of the recently sequenced lamprey germline genome identified two CR sequences, CR1 and CR2, containing the 344 previously un-identified amino acids at the amino terminus. CR1 also contains a novel four amino acid insertion in the DNA-binding domain (DBD). We studied corticosteroid activation of CR1 and CR2 and found their strongest response was to 11-deoxycorticosterone and 11-deoxycortisol, the two circulating corticosteroids in lamprey. Based on steroid specificity, both CRs are close to elephant shark MR and distant from elephant shark GR. HEK293 cells transfected with full-length CR1 or CR2 and the MMTV promoter have about 3-fold higher steroid-mediated activation compared to HEK293 cells transfected with these CRs and the TAT3 promoter. Deletion of the amino-terminal domain (NTD) of lamprey CR1 and CR2 to form truncated CRs decreased transcriptional activation by about 70% in HEK293 cells transfected with MMTV, but increased transcription by about 6-fold in cells transfected with TAT3, indicating that the promoter has an important effect on NTD regulation of CR transcription by corticosteroids.