BAX and BAK1 are dispensable for ABT-737-induced dissociation of the BCL2-BECN1 complex and autophagy

Autophagy. 2015;11(3):452-9. doi: 10.1080/15548627.2015.1017191.

Abstract

Disruption of the complex of BECN1 with BCL2 or BCL2L1/BCL-XL is an essential switch that turns on cellular autophagy in response to environmental stress or treatment with BH3 peptidomimetics. Recently, it has been proposed that BCL2 and BCL2L1/BCL-XL may inhibit autophagy indirectly through a mechanism dependent on the proapoptotic BCL2 family members, BAX and BAK1. Here we report that the BH3 mimetic, ABT-737, induces autophagy in parallel with disruption of BCL2-BECN1 binding in 2 different apoptosis-deficient cell types lacking BAX and BAK1, namely in mouse embryonic fibroblasts cells and in human colon cancer HCT116 cells. We conclude that the BH3 mimetic ABT-737 induces autophagy through a BAX and BAK1-independent mechanism that likely involves disruption of BECN1 binding to antiapoptotic BCL2 family members.

Keywords: ABT-737; ACTB, actin, β; BAK1; BAK1, BCL2-antagonist/killer 1; BAX; BAX, BCL2-associated X protein; BCL2; BCL2, B-cell CLL/lymphoma 2; BECN1 (Beclin 1); BECN1, Beclin 1, autophagy-related; Baf A1, bafilomycin A1; DKO, double-knockout; FBS, fetal bovine serum; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HBSS, Hanks’ balanced salt solution; HRP, horseradish peroxidase; MAP1LC3/LC3, microtubule-associated protein 1 light chain 3; MCL1, myeloid cell leukemia 1; MEFs, mouse embryonic fibroblasts; MTOR, mechanistic target of rapamycin; PBS, phosphate-buffered saline; SQSTM1, sequestosome 1; STS, staurosporine; WT, wild type; apoptosis; autophagy.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Apoptosis Regulatory Proteins / metabolism*
  • Autophagy*
  • Beclin-1
  • Biphenyl Compounds / chemistry*
  • Cell Line, Tumor
  • Fibroblasts / metabolism
  • Flow Cytometry
  • HCT116 Cells
  • Humans
  • Membrane Proteins / metabolism*
  • Mice
  • Mice, Knockout
  • Microscopy, Fluorescence
  • Nitrophenols / chemistry*
  • Peptide Fragments
  • Piperazines / chemistry
  • Protein Binding
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-bcl-2 / metabolism*
  • Sulfonamides / chemistry*
  • bcl-2 Homologous Antagonist-Killer Protein / metabolism*
  • bcl-2-Associated X Protein / metabolism*

Substances

  • ABT-737
  • Apoptosis Regulatory Proteins
  • BAK1 protein, human
  • BAX protein, human
  • BCL2 protein, human
  • BECN1 protein, human
  • Bak1 protein, mouse
  • Bax protein (53-86)
  • Bax protein, mouse
  • Beclin-1
  • Becn1 protein, mouse
  • Biphenyl Compounds
  • Membrane Proteins
  • Nitrophenols
  • Peptide Fragments
  • Piperazines
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • Sulfonamides
  • bcl-2 Homologous Antagonist-Killer Protein
  • bcl-2-Associated X Protein
  • Bcl2 protein, mouse