Abstract
Vitamin D receptor (VDR) is a substrate for modification with small ubiquitin-like modifier (SUMO). To further assess the role of reversible SUMOylation within the vitamin D hormonal response, we evaluated the effects of sentrin/SUMO-specific proteases (SENPs) that can function to remove small ubiquitin-like modifier (SUMO) from target proteins upon the activities of VDR and related receptors. We report that SENP1 and SENP2 strikingly potentiate ligand-mediated transactivation of VDR and also its heterodimeric partner, retinoid X receptor (RXRα) with depletion of cellular SENP1 significantly diminishing the hormonal responsiveness of the endogenous vitamin D target gene CYP24A1. We find that SENP-directed modulation of VDR activity is cell line-dependent, achieving potent modulatory effects in Caco-2 and HEK-293 cells, while in MCF-7 cells the vitamin D signal is unaffected by any tested SENP. In support of their function as novel modulators of the vitamin D hormonal pathway we demonstrate that both SENP1 and SENP2 can interact with VDR and reverse its modification with SUMO2. In a preliminary analysis we identify lysine 91, a residue known to be critical for formation and DNA binding of the VDR-RXR heterodimer, as a minor SUMO acceptor site within VDR. In combination, our results support a repressor function for SUMOylation of VDR and reveal SENPs as a novel class of VDR/RXR co-regulatory protein that significantly modulate the vitamin D response and which could also have important impact upon the functionality of both RXR-containing homo and heterodimers.
Original language | English |
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Pages (from-to) | e89506 |
Journal | PLoS ONE |
Volume | 9 |
Issue number | 2 |
DOIs | |
Publication status | Published - 2014 |
Externally published | Yes |
Keywords
- Animals
- Blotting, Western
- CHO Cells
- Cricetulus
- Cysteine Endopeptidases/genetics
- Endopeptidases/genetics
- Gene Expression Regulation
- HEK293 Cells
- Humans
- MCF-7 Cells
- Mutagenesis, Site-Directed
- Mutation/genetics
- RNA, Messenger/genetics
- Real-Time Polymerase Chain Reaction
- Receptors, Calcitriol/genetics
- Retinoid X Receptors/genetics
- Reverse Transcriptase Polymerase Chain Reaction
- Signal Transduction
- Small Ubiquitin-Related Modifier Proteins/genetics
- Transcription, Genetic