A Small RNA Derived from RNA Coactivator SRA Blocks Steroid Receptor Signaling via Inhibition of Pus1p-Mediated Pseudouridylation of SRA: Evidence of a Novel RNA Binding Domain in the N-Terminus of Steroid Receptors

Estrogen receptors (ERs) and androgen receptors (ARs) are important targets for cancer therapy; however, the efficacy of receptor antagonists is limited, and alternative strategies are needed. Steroid receptor RNA Activator (SRA) is a long, noncoding RNA coactivator (although some protein-encoding 5' splice variants have also been reported) that requires pseudouridylation by Pus1p to stimulate steroid receptor signaling. A uridine at position 206 (U206), which is located in small hairpin structure STR5 in the conserved SRA core sequence, is a critical target for pseudouridylation. We assessed if synthetic STR5 could serve as a novel competitive inhibitor of ERα and AR signaling by disrupting the Pus1p-SRA-steroid receptor axis. STR5 specifically inhibited Pus1p-dependent pseudouridylation of SRA with higher efficiency than STR5 mutant U206A. We show that SRA binds to the N-terminal domain (NTD) of ERα and AR with high affinity despite the absence of a recognizable RNA binding motif (RBM). Finally, we show that STR5 specifically inhibits ERα- and AR-dependent transactivation of target genes in steroid-sensitive cancer cells, consistent with disruption of the targeted Pus1p-SRA pathway. Together, our results show that the NTD of ERα and AR contains a novel RBM that directly binds SRA, and that STR5 can serve as a novel class of RNA inhibitor of ERα and AR signaling by interfering with Pus1p-mediated SRA pseudouridylation. Targeting this unexplored receptor signaling pathway may pave the way for the development of new types of cancer therapeutics.