Abstract
The inositol 1,4,5-trisphosphate receptor (IP3R) is a major regulator of apoptotic signaling. Through interactions with members of the Bcl-2 family of proteins, it drives calcium (Ca2+) transients from the endoplasmic reticulum (ER) to mitochondria, thereby establishing a functional and physical link between these organelles. Importantly, the IP3R also regulates autophagy, and in particular, its inhibition/depletion strongly induces macroautophagy. Here, we show that the IP3R antagonist xestospongin B induces autophagy by disrupting a molecular complex formed by the IP3R and Beclin 1, an interaction that is increased or inhibited by overexpression or knockdown of Bcl-2, respectively. An effect of Beclin 1 on Ca2+ homeostasis was discarded as siRNA-mediated knockdown of Beclin 1 did not affect cytosolic or luminal ER Ca2+ levels. Xestospongin B- or starvation-induced autophagy was inhibited by overexpression of the IP3R ligand-binding domain, which coimmunoprecipitated with Beclin 1. These results identify IP3R as a new regulator of the Beclin 1 complex that may bridge signals converging on the ER and initial phagophore formation.
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Abbreviations
- ACD:
-
autophagic cell death
- Ambra:
-
activating molecule in Beclin 1-regulated autophagy
- Atg:
-
autophagy-related gene
- Ca2+:
-
calcium
- [Ca2+]:
-
Ca2+ concentration
- [Ca2+]c:
-
cytosolic [Ca2+]
- CFP:
-
cyan fluorescent protein
- cytAEQ:
-
cytosolic aequorin
- Cyto:
-
cytosolic
- ER:
-
endoplasmic reticulum
- FBS:
-
fetal bovine serum
- FRET:
-
fluorescence resonance energy transfer
- GFP:
-
green fluorescent protein
- GFP-LC3:
-
GFP-coupled microtubule-associated protein light chain 3
- IMP:
-
inositol monophosphatase
- IP3:
-
myo-inositol 1,4,5-trisphosphate
- IP3R:
-
IP3 receptor
- LBD:
-
ligand-binding domain
- mTOR:
-
mammalian target of rapamycin
- OMM:
-
outer mitochondrial membrane
- PI3K:
-
phosphatidylinositol 3-kinase
- PI3P:
-
phosphatidylinositol-3-phosphate
- RFP:
-
red fluorescent protein
- SERCA:
-
sarco–endoplasmic reticulum Ca2+ ATPase
- tBHQ:
-
2,5-di(ter-butyl)-1,4 benzohydroquinone
- UVRAG:
-
UV irradiation resistance-associated tumor suppressor gene
- Vps:
-
vacuolar protein sorting
- YFP:
-
yellow fluorescent protein
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Acknowledgements
GK is supported by the Ligue Nationale contre le Cancer (équipe labellisée), European Commission (RIGHT, ChemoRes, Apop-Train, Apo-SYs), Agence Nationale pour la Recherche, Cancéropôle Ile-de-France and Institut National contre le Cancer (INCa). We thank the International Collaboration Program ECOS-CONICYT, project C08S01. JMV is supported by Fondation pour la Recherche Médicale and CONICYT. CO holds a PhD scholarship from CONICYT, Chile. OK is supported by EMBO. AWEJ is a Medical Research Council student. We also thank Michael R Duchen (University College London) for the use of confocal facility (Zeiss LSM 510 Meta) and discussions, as well as to Dr. Roger Y Tsien (University of California at San Diego) for the donation of ERD1.
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Vicencio, J., Ortiz, C., Criollo, A. et al. The inositol 1,4,5-trisphosphate receptor regulates autophagy through its interaction with Beclin 1. Cell Death Differ 16, 1006–1017 (2009). https://doi.org/10.1038/cdd.2009.34
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DOI: https://doi.org/10.1038/cdd.2009.34
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