Direct transactivation of the anti-apoptotic gene apolipoprotein J (clusterin) by B-MYB

J Biol Chem. 2000 Jul 14;275(28):21055-60. doi: 10.1074/jbc.M002055200.

Abstract

B-MYB is a ubiquitously expressed transcription factor involved in the regulation of cell survival, proliferation, and differentiation. In an attempt to isolate B-MYB-regulated genes that may explain the role of B-MYB in cellular processes, representational difference analysis was performed in neuroblastoma cell lines with different levels of B-MYB expression. One of the genes, the mRNA levels of which were enhanced in B-MYB expressing cells, was ApoJ/Clusterin(SGP-2/TRMP-2) (ApoJ/Clusterin), previously implicated in regulation of apoptosis and tumor progression. Here we show that the human ApoJ/Clusterin gene contains a Myb binding site in its 5' flanking region, which interacts with bacterially synthesized B-MYB protein and mediates B-MYB-dependent transactivation of the ApoJ/Clusterin promoter in transient transfection assays. Endogenous ApoJ/Clusterin expression is induced in mammalian cell lines following transient transfection of a B-MYB cDNA. Blockage of secreted clusterin by a monoclonal antibody results in increased apoptosis of neuroblastoma cells exposed to the chemotherapeutic drug doxorubicin. Thus, activation of ApoJ/Clusterin by B-MYB may be an important step in the regulation of apoptosis in normal and diseased cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • COS Cells
  • Cell Cycle Proteins*
  • Clusterin
  • DNA-Binding Proteins / metabolism*
  • Glycoproteins / biosynthesis
  • Glycoproteins / genetics*
  • Humans
  • Molecular Chaperones*
  • Molecular Sequence Data
  • Neoplasm Proteins / genetics
  • Neuroblastoma
  • Oncogene Proteins / metabolism
  • Promoter Regions, Genetic*
  • RNA, Messenger / genetics
  • Rats
  • Recombinant Proteins / biosynthesis
  • Sequence Alignment
  • Sequence Homology, Nucleic Acid
  • Trans-Activators / metabolism*
  • Transcription, Genetic*
  • Transcriptional Activation*
  • Transfection
  • Tumor Cells, Cultured

Substances

  • CLU protein, human
  • Cell Cycle Proteins
  • Clusterin
  • DNA-Binding Proteins
  • Glycoproteins
  • MYBL2 protein, human
  • Molecular Chaperones
  • Neoplasm Proteins
  • Oncogene Proteins
  • RNA, Messenger
  • Recombinant Proteins
  • Trans-Activators