Abstract
Retinoic acid receptor-related orphan receptor alpha (RORα), a candidate tumor suppressor, is prevalently downregulated or lost in malignant breast cancer cells. However, the mechanisms of how RORα expression is regulated in breast epithelial cells remain incompletely understood. Protein arginine N-methyltransferase 5 (PRMT5), a type II methyltransferase catalyzing the symmetric methylation of the amino acid arginine in target proteins, was reported to regulate protein stability. To study whether and how PRMT5 regulates RORα, we examined the direct interaction between RORα and PRMT5 by immunoprecipitation and GST pull-down assays. The results showed that PRMT5 directly bound to RORα, and PRMT5 mainly symmetrically dimethylated the DNA-binding domain (DBD) but not the ligand-binding domain (LBD) of RORα. To investigate whether RORα protein stability is regulated by PRMT5, we transfected HEK293FT cells with RORα and PRMT5- expressing or PRMT5-silencing (shPRMT5) vectors and then examined RORα protein stability by a cycloheximide chase assay. The results showed that PRMT5 increased RORα protein stability, while silencing PRMT5 accelerated RORα protein degradation. In PRMT5-silenced mammary epithelial cells, RORα protein expression was decreased, accompanied by an enhanced epithelial–mesenchymal transition morphology and cell invasion and migration abilities. In PRMT5-overexpressed mammary epithelial cells, RORα protein was accumulated, and cell invasion was suppressed. These findings revealed a novel mechanism by which PRMT5 regulates RORα protein stability.
Document Type
Article
Publication Date
5-2024
Digital Object Identifier (DOI)
https://doi.org/10.3390/cancers16101914
Funding Information
This research was supported by the National Institutes of Health (grant nos.: 1R01CA207772, 1R01 CA277946 and 1R01CA215095 to R.X. and 7R21AG080176 to G.X.) and the American Cancer Society (grant no.: IRG-16-182-28 to G.X.).
Repository Citation
Xiong, Gaofeng; Obringer, Brynne; Jones, Austen; Horton, Elise; and Xu, Ren, "Regulation of RORα Stability through PRMT5-Dependent Symmetric Dimethylation" (2024). Markey Cancer Center Faculty Publications. 256.
https://uknowledge.uky.edu/markey_facpub/256
Notes/Citation Information
© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).