Inhibition of glucocorticoid-induced apoptosis in 697 pre-B lymphocytes by the mineralocorticoid receptor N-terminal domain

J Biol Chem. 2002 Nov 1;277(44):42188-96. doi: 10.1074/jbc.M205085200. Epub 2002 Aug 22.

Abstract

The glucocorticoid and mineralocorticoid receptors (GR and MR) share considerable structural and functional homology and bind as homodimers to hormone-response elements. We have shown previously that MR and GR can form heterodimers that inhibit transcription from a glucocorticoid (GC)-responsive gene and that this inhibition was mediated by the N-terminal domain (NTD) of MR. In this report, we examined the effect of NTD-MR on GC-induced apoptosis in the GC-sensitive pre-B lymphoma cell line, 697. In GC-treated 697 cells, we demonstrated that stable expression of NTD-MR blocks apoptosis and inhibits proteolytic processing of pro-caspases-3, -8, and -9 and poly(ADP-ribose) polymerase. Importantly, gel shift and immunoprecipitation analyses revealed a direct association between the GR and amino acids 203-603 of NTD-MR. We observed down-regulation of c-Myc and of the anti-apoptotic proteins Bcl-2 and Bfl-1 as well as high levels of the pro-apoptotic proteins Bax and Bid. Conversely, cells stably expressing NTD-MR exhibited increased expression of Bcl-2 and Bfl-1 and diminished levels of Bid and Bax. These data provide a potential mechanism for the observed inhibition of cytochrome c and Smac release from the mitochondria of NTD-MR cells and resultant resistance to GC-induced apoptosis. Thus, NTD-MR may mediate GC effects through heterodimerization with GR and ensuing inhibition of GC-regulated gene transcription.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Active Transport, Cell Nucleus
  • Apoptosis / drug effects*
  • Apoptosis Regulatory Proteins
  • Carrier Proteins / metabolism
  • Caspases / metabolism
  • Cytochrome c Group / metabolism
  • Dimerization
  • Glucocorticoids / antagonists & inhibitors*
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Minor Histocompatibility Antigens
  • Mitochondrial Proteins / metabolism
  • Peptide Fragments / physiology*
  • Poly(ADP-ribose) Polymerases / metabolism
  • Precursor B-Cell Lymphoblastic Leukemia-Lymphoma / pathology*
  • Proto-Oncogene Proteins c-bcl-2 / analysis
  • Receptors, Mineralocorticoid / physiology*
  • Transcription, Genetic
  • Tumor Cells, Cultured

Substances

  • Apoptosis Regulatory Proteins
  • BCL2-related protein A1
  • Carrier Proteins
  • Cytochrome c Group
  • DIABLO protein, human
  • Glucocorticoids
  • Intracellular Signaling Peptides and Proteins
  • Minor Histocompatibility Antigens
  • Mitochondrial Proteins
  • Peptide Fragments
  • Proto-Oncogene Proteins c-bcl-2
  • Receptors, Mineralocorticoid
  • Poly(ADP-ribose) Polymerases
  • Caspases