Proteasome-mediated degradation of the vitamin D receptor (VDR) and a putative role for SUG1 interaction with the AF-2 domain of VDR

Hisashi Masuyama, Paul N. MacDonald

Research output: Contribution to journalArticle

97 Citations (Scopus)

Abstract

The AF-2 helix of nuclear receptors is essential for ligand-activated transcription, and it may function to couple the receptor to transcriptional coactivator proteins. This domain also contacts components of the proteasome machinery, suggesting that nuclear receptors may be targets for proteasome- mediated proteolysis. In the present study, we demonstrate that mSUG1 (P45), a component of the 265 proteasome, interacts in a 1,25-(OH)2D3-dependent manner with the AF-2 domain of the vitamin D receptor (VDR). Furthermore, treatment of ROS 17/2.8 osteosarcoma cells with the proteasome inhibitors MG132 or β-lactone increased steady-state levels of the VDR protein. In the presence cycloheximide (10 μg/ml), the liganded VDR protein was degraded with a half-life of approximately 8 h, and this rate of degradation was completely blocked by 0.05 mM MG132. The role of SUG1-VDR interaction in this process was investigated in transient expression studies. Overexpression of wild-type mSUG1 in ROS17/2.8 cells generated a novel proteolytic VDR fragment of approximately 50 kDa, and its production was blocked by proteasome inhibitors or by a nonhydrolyzable ATP analog. Parallel studies with SUG1 (K196H), a mutant that does not interact with the VDR, did not produce the 50 kDa VDR fragment. Functionally, expression of SUG1 in a VDR-responsive reporter gene assay resulted in a profound inhibition of 1,25-(OH)2D3- activated transcription, while expression of SUG1(K196H) had no significant effect in this system. These data show that the AF-2 domain of VDR interacts with SUG1 in a 1,25-(OH)2D3-dependent fashion and that this interaction may target VDR to proteasome-mediated degradation as a means to down-regulate the 1,25-(OH)2D3-activated transcriptional response.

Original languageEnglish
Pages (from-to)429-440
Number of pages12
JournalJournal of Cellular Biochemistry
Volume71
Issue number3
DOIs
Publication statusPublished - Dec 1 1998
Externally publishedYes

Keywords

  • 1,25-(OH)D
  • AF-2 domain
  • Proteasome
  • SUG 1
  • VDR

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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