Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) protein-induced lysosomal translocation of proapoptotic effectors is mediated by phosphofurin acidic cluster sorting protein-2 (PACS-2)

Nathan W. Werneburg, Steve F. Bronk, Maria Eugenia Guicciardi, Laurel Thomas, Jimmy D. Dikeakos, Gary Thomas, Gregory James Gores

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Abstract

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis of liver cancer cell lines requires death receptor-5 (DR5)-dependent permeabilization of lysosomal membranes. Ligated DR5 triggers recruitment of the proapoptotic proteins Bim and Bax to lysosomes, releasing cathepsin B into the cytosol where it mediates mitochondria membrane permeabilization and activation of executioner caspases. Despite the requirement for lysosome membrane permeabilization during TRAIL-induced apoptosis, little is known about the mechanism that controls recruitment of Bim and Bax to lysosomal membranes. Here we report that TRAIL induces recruitment of the multifunctional sorting protein phosphofurin acidic cluster sorting protein-2 (PACS-2) to DR5-positive endosomes in Huh-7 cells where it forms an immunoprecipitatable complex with Bim and Bax on lysosomal membranes. shRNA-targeted knockdown of PACS-2 prevents recruitment of Bim or Bax to lysosomes, blunting the TRAIL-induced lysosome membrane permeabilization. Consistent with the reduced lyso-some membrane permeabilization, shRNA knockdown of PACS-2 in Huh-7 cells reduced TRAIL-induced apoptosis and increased clonogenic cell survival. The determination that recombinant PACS-2 bound Bim but not Bax in vitro and that shRNA knockdown of Bim blocked Bax recruitment to lyso-somes suggests that TRAIL/DR5 triggers endosomal PACS-2 to recruit Bim and Bax to lysosomes to release cathepsin B and induce apoptosis. Together, these findings provide insight into the lysosomal pathway of apoptosis.

Original languageEnglish (US)
Pages (from-to)24427-24437
Number of pages11
JournalJournal of Biological Chemistry
Volume287
Issue number29
DOIs
StatePublished - Jul 13 2012

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TNF-Related Apoptosis-Inducing Ligand
Protein Transport
TNF-Related Apoptosis-Inducing Ligand Receptors
Sorting
Tumor Necrosis Factor-alpha
Lysosomes
Apoptosis
Ligands
Membranes
Proteins
Small Interfering RNA
Cathepsin B
Cells
bcl-2-Associated X Protein
Mitochondria
Endosomes
Liver Neoplasms
Caspases
Liver
Cytosol

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) protein-induced lysosomal translocation of proapoptotic effectors is mediated by phosphofurin acidic cluster sorting protein-2 (PACS-2). / Werneburg, Nathan W.; Bronk, Steve F.; Guicciardi, Maria Eugenia; Thomas, Laurel; Dikeakos, Jimmy D.; Thomas, Gary; Gores, Gregory James.

In: Journal of Biological Chemistry, Vol. 287, No. 29, 13.07.2012, p. 24427-24437.

Research output: Contribution to journalArticle

Werneburg, Nathan W. ; Bronk, Steve F. ; Guicciardi, Maria Eugenia ; Thomas, Laurel ; Dikeakos, Jimmy D. ; Thomas, Gary ; Gores, Gregory James. / Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) protein-induced lysosomal translocation of proapoptotic effectors is mediated by phosphofurin acidic cluster sorting protein-2 (PACS-2). In: Journal of Biological Chemistry. 2012 ; Vol. 287, No. 29. pp. 24427-24437.
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