p53 at the endoplasmic reticulum regulates apoptosis in a Ca²⁺-dependent manner

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 112; no. 6; pp. 1779 - 1784
• Main Authors: Giorgi, Carlotta, Bonora, Massimo, Sorrentino, Giovanni, Missiroli, Sonia, Poletti, Federica, Suski, Jan M., Ramirez, Fabian Galindo, Rizzuto, Rosario, Di Virgilio, Francesco, Zito, Ester, Pandolfi, Pier Paolo, Wieckowski, Mariusz R., Mammano, Fabio, Del Sal, Giannino, Pinton, Paolo
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 10-02-2015; National Acad Sciences
• Subjects: Aequorin - metabolism; Animals; Apoptosis - physiology; Biological Sciences; Blotting, Western; Calcium - metabolism; Cell Line; Cytosol - metabolism; Endoplasmic Reticulum - metabolism; Flow Cytometry; Fluorescence Resonance Energy Transfer; Fura-2; Gene Knockdown Techniques; Humans; Immunoprecipitation; Mice; Mitochondria - metabolism; Sarcoplasmic Reticulum Calcium-Transporting ATPases - metabolism; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - metabolism; mitochondria-associated membranes; apoptosis; calcium; endoplasmic reticulum; p53
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1410723112

Significance Accumulating evidence has underscored the role of cytosolic p53 in promoting cell death. Different reports have revealed that p53 participates in apoptosis induction by acting directly at mitochondria. However, because p53 can mediate apoptosis without its DNA-binding domain (the domain proposed to be fundamental for the targeting of p53 to mitochondria), the mitochondrial localization of p53 is likely not the only transcription-independent mechanism by which p53 promotes apoptosis. Here we demonstrate that p53 at the endoplasmic reticulum (ER) and at mitochondria-associated membranes, interacting with sarco/ER Ca ²⁺-ATPase pumps, modulates ER–mitochondria cross-talk and, in turn, Ca ²⁺-dependent apoptosis.
The tumor suppressor p53 is a key protein in preventing cell transformation and tumor progression. Activated by a variety of stimuli, p53 regulates cell-cycle arrest and apoptosis. Along with its well-documented transcriptional control over cell-death programs within the nucleus, p53 exerts crucial although still poorly understood functions in the cytoplasm, directly modulating the apoptotic response at the mitochondrial level. Calcium (Ca ²⁺) transfer between the endoplasmic reticulum (ER) and mitochondria represents a critical signal in the induction of apoptosis. However, the mechanism controlling this flux in response to stress stimuli remains largely unknown. Here we show that, in the cytoplasm, WT p53 localizes at the ER and at specialized contact domains between the ER and mitochondria (mitochondria-associated membranes). We demonstrate that, upon stress stimuli, WT p53 accumulates at these sites and modulates Ca ²⁺ homeostasis. Mechanistically, upon activation, WT p53 directly binds to the sarco/ER Ca ²⁺-ATPase (SERCA) pump at the ER, changing its oxidative state and thus leading to an increased Ca ²⁺ load, followed by an enhanced transfer to mitochondria. The consequent mitochondrial Ca ²⁺ overload causes in turn alterations in the morphology of this organelle and induction of apoptosis. Pharmacological inactivation of WT p53 or naturally occurring p53 missense mutants inhibits SERCA pump activity at the ER, leading to a reduction of the Ca ²⁺ signaling from the ER to mitochondria. These findings define a critical nonnuclear function of p53 in regulating Ca ²⁺ signal-dependent apoptosis.