BH3-only proteins target BCL-xL/MCL-1, not BAX/BAK, to initiate apoptosis

Cell Res. 2019 Nov;29(11):942-952. doi: 10.1038/s41422-019-0231-y. Epub 2019 Sep 24.

Abstract

It has been widely accepted that mitochondria-dependent apoptosis initiates when select BH3-only proteins (BID, BIM, etc.) directly engage and allosterically activate effector proteins BAX/BAK. Here, through reconstitution of cells lacking all eight pro-apoptotic BH3-only proteins, we demonstrate that all BH3-only proteins primarily target the anti-apoptotic BCL-2 proteins BCL-xL/MCL-1, whose simultaneous suppression enables membrane-mediated spontaneous activation of BAX/BAK. BH3-only proteins' apoptotic activities correlate with affinities for BCL-xL/MCL-1 instead of abilities to directly activate BAX/BAK. Further, BID and BIM do not distinguish BAX from BAK or accelerate BAX/BAK activation following inactivation of BCL-xL/MCL-1. Remarkably, death ligand-induced apoptosis in cells lacking BH3-only proteins and MCL-1 is fully restored by BID mutants capable of neutralizing BCL-xL, but not direct activation of BAX/BAK. Taken together, our findings provide a "Membrane-mediated Permissive" model, in which the BH3-only proteins only indirectly activate BAX/BAK by neutralizing the anti-apoptotic BCL-2 proteins, and thus allowing BAX/BAK to undergo unimpeded, spontaneous activation in the mitochondrial outer membrane milieu, leading to apoptosis initiation.

MeSH terms

  • Apoptosis / physiology*
  • BH3 Interacting Domain Death Agonist Protein / physiology*
  • Bcl-2-Like Protein 11 / physiology*
  • HCT116 Cells
  • HEK293 Cells
  • Humans
  • Mitochondria / metabolism*
  • Mitochondrial Membranes / metabolism*
  • Myeloid Cell Leukemia Sequence 1 Protein / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • bcl-2 Homologous Antagonist-Killer Protein / metabolism
  • bcl-X Protein / metabolism

Substances

  • BAK1 protein, human
  • BCL2 protein, human
  • BCL2L1 protein, human
  • BH3 Interacting Domain Death Agonist Protein
  • BID protein, human
  • Bcl-2-Like Protein 11
  • MCL1 protein, human
  • Myeloid Cell Leukemia Sequence 1 Protein
  • Proto-Oncogene Proteins c-bcl-2
  • bcl-2 Homologous Antagonist-Killer Protein
  • bcl-X Protein