Apoptosis

, Volume 17, Issue 8, pp 810–820 | Cite as

Cleavage of Atg3 protein by caspase-8 regulates autophagy during receptor-activated cell death

  • Ozlem Oral
  • Devrim Oz-Arslan
  • Zeynep Itah
  • Atabak Naghavi
  • Remziye Deveci
  • Sabire Karacali
  • Devrim Gozuacik
Original Paper

Abstract

Autophagy is an evolutionarily conserved mechanism contributing to cell survival under stress conditions including nutrient and growth factor deprivation. Connections and cross-talk between cell death mechanisms and autophagy is under investigation. Here, we describe Atg3, an essential regulatory component of autophagosome biogenesis, as a new substrate of caspase-8 during receptor-mediated cell death. Both, tumor necrosis factor α and tumor necrosis factor-related apoptosis inducing ligand induced cell death was accompanied by Atg3 cleavage and this event was inhibited by a pan-caspase inhibitor (zVAD) or a caspase-8-specific inhibitor (zIETD). Indeed, caspase-8 overexpression led to Atg3 degradation and this event depended on caspase-8 enzymatic activity. Mutation of the caspase-8 cleavage site on Atg3 abolished its cleavage both in vitro and in vivo, demonstrating that Atg3 was a direct target of caspase-8. Autophagy was inactive during apoptosis and blockage of caspases or overexpression of a non-cleavable Atg3 protein reestablished autophagic activity upon death receptor stimulation. In this system, autophagy was important for cell survival since inhibition of autophagy increased cell death. Therefore, Atg3 provides a novel link between apoptosis and autophagy during receptor-activated cell death.

Keywords

Apoptosis Autophagy Death receptor TNF-α TRAIL Caspase-8 Cell survival 

Abbreviations

LC3

Microtubule-associated protein 1 (MAP1) light chain 3

PI3K

Phosphatidylinositol 3-kinase

zVAD-fmk

Benzyloxycarbonyl-valyl-alanyl-aspartic-acid (O-methyl)-fluoromethylketone

CHX

Cycloheximide

TNF

Tumor necrosis factor

TRAIL

Tumor necrosis factor-related apoptosis-inducing ligand

FADD

Fas-associated protein with death domain

TRADD

Tumor necrosis factor receptor type 1-associated death domain protein

PE

Phosphatidylethanolamine

HEK293T

Human embryonic kidney cells

Supplementary material

10495_2012_735_MOESM1_ESM.pdf (382 kb)
Supplementary material 1 (PDF 382 kb)
10495_2012_735_MOESM2_ESM.doc (26 kb)
Supplementary material 2 (DOC 25 kb)

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Ozlem Oral
    • 1
  • Devrim Oz-Arslan
    • 1
    • 2
  • Zeynep Itah
    • 1
  • Atabak Naghavi
    • 3
  • Remziye Deveci
    • 3
  • Sabire Karacali
    • 3
  • Devrim Gozuacik
    • 1
  1. 1.Faculty of Engineering and Natural Sciences, Biological Sciences and Bioengineering ProgramSabanci UniversityIstanbulTurkey
  2. 2.Faculty of Medicine, Department of BiophysicsAcibadem UniversityIstanbulTurkey
  3. 3.Faculty of Science, Department of BiologyEge UniversityIzmirTurkey

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