Bcl-2 Prevents Apoptotic Mitochondrial Dysfunction by Regulating Proton Flux

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1998 Mar 21

PMID 9465036

Citations 78

Authors

S Shimizu

Y Eguchi

W Kamiike

Y Funahashi

A Mignon

V Lacronique

H Matsuda

Y Tsujimoto

Affiliations

Soon will be listed here.

Abstract

We and others have recently shown that loss of the mitochondrial membrane potential (Deltapsi) precedes apoptosis and chemical-hypoxia-induced necrosis and is prevented by Bcl-2. In this report, we examine the biochemical mechanism used by Bcl-2 to prevent Deltapsi loss, as determined with mitochondria isolated from a cell line overexpressing human Bcl-2 or from livers of Bcl-2 transgenic mice. Although Bcl-2 had no effect on the respiration rate of isolated mitochondria, it prevented both Deltapsi loss and the permeability transition (PT) induced by various reagents, including Ca2+, H2O2, and tert-butyl hydroperoxide. Even under conditions that did not allow PT, Bcl-2 maintained Deltapsi, suggesting that the functional target of Bcl-2 is regulation of Deltapsi but not PT. Bcl-2 also maintained Deltapsi in the presence of the protonophore SF6847, which induces proton influx, suggesting that Bcl-2 regulates ion transport to maintain Deltapsi. Although treatment with SF6847 in the absence of Ca2+ caused massive H+ influx in control mitochondria, the presence of Bcl-2 induced H+ efflux after transient H+ influx. In this case, Bcl-2 did not enhance K+ efflux. Furthermore, Bcl-2 enhanced H+ efflux but not K+ flux after treatment of mitochondria with Ca2+ or tert-butyl hydroperoxide. These results suggest that Bcl-2 maintains Deltapsi by enhancing H+ efflux in the presence of Deltapsi-loss-inducing stimuli.

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References

Wu D, Wallen H, Nunez G . Interaction and regulation of subcellular localization of CED-4 by CED-9. Science. 1997; 275(5303):1126-9. DOI: 10.1126/science.275.5303.1126. View

Kluck R, Bossy-Wetzel E, Green D, Newmeyer D . The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis. Science. 1997; 275(5303):1132-6. DOI: 10.1126/science.275.5303.1132. View

Monaghan P, Robertson D, Amos T, Dyer M, Mason D, Greaves M . Ultrastructural localization of bcl-2 protein. J Histochem Cytochem. 1992; 40(12):1819-25. DOI: 10.1177/40.12.1453000. View

Chittenden T, Harrington E, OConnor R, Flemington C, Lutz R, Evan G . Induction of apoptosis by the Bcl-2 homologue Bak. Nature. 1995; 374(6524):733-6. DOI: 10.1038/374733a0. View

Kohga S, Kinjo M, Tanaka K, Ogawa H, Ishihara M, Tanaka N . Effects of 5-(2-chlorobenzyl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridine hydrochloride (Ticlopidine), a platelet aggregation inhibitor, on blood-borne metastasis. Cancer Res. 1981; 41(11 Pt 1):4710-4. View

Vaux D, Cory S, Adams J . Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B cells. Nature. 1988; 335(6189):440-2. DOI: 10.1038/335440a0. View

Hengartner M, Horvitz H . C. elegans cell survival gene ced-9 encodes a functional homolog of the mammalian proto-oncogene bcl-2. Cell. 1994; 76(4):665-76. DOI: 10.1016/0092-8674(94)90506-1. View

Shimizu S, Eguchi Y, Kamiike W, Waguri S, Uchiyama Y, Matsuda H . Retardation of chemical hypoxia-induced necrotic cell death by Bcl-2 and ICE inhibitors: possible involvement of common mediators in apoptotic and necrotic signal transductions. Oncogene. 1996; 12(10):2045-50. View

Yang J, Liu X, Bhalla K, Kim C, Ibrado A, Cai J . Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science. 1997; 275(5303):1129-32. DOI: 10.1126/science.275.5303.1129. View

10.

Nakai M, Takeda A, Cleary M, Endo T . The bcl-2 protein is inserted into the outer membrane but not into the inner membrane of rat liver mitochondria in vitro. Biochem Biophys Res Commun. 1993; 196(1):233-9. DOI: 10.1006/bbrc.1993.2239. View

11.

Spector M, Desnoyers S, Hoeppner D, Hengartner M . Interaction between the C. elegans cell-death regulators CED-9 and CED-4. Nature. 1997; 385(6617):653-6. DOI: 10.1038/385653a0. View

12.

Cleary M, Sklar J . Nucleotide sequence of a t(14;18) chromosomal breakpoint in follicular lymphoma and demonstration of a breakpoint-cluster region near a transcriptionally active locus on chromosome 18. Proc Natl Acad Sci U S A. 1985; 82(21):7439-43. PMC: 391360. DOI: 10.1073/pnas.82.21.7439. View

13.

Bakhshi A, Jensen J, Goldman P, Wright J, MCBRIDE O, Epstein A . Cloning the chromosomal breakpoint of t(14;18) human lymphomas: clustering around JH on chromosome 14 and near a transcriptional unit on 18. Cell. 1985; 41(3):899-906. DOI: 10.1016/s0092-8674(85)80070-2. View

14.

Shimizu S, Eguchi Y, Kamiike W, Waguri S, Uchiyama Y, Matsuda H . Bcl-2 blocks loss of mitochondrial membrane potential while ICE inhibitors act at a different step during inhibition of death induced by respiratory chain inhibitors. Oncogene. 1996; 13(1):21-9. View

15.

Eguchi Y, Ewert D, Tsujimoto Y . Isolation and characterization of the chicken bcl-2 gene: expression in a variety of tissues including lymphoid and neuronal organs in adult and embryo. Nucleic Acids Res. 1992; 20(16):4187-92. PMC: 334124. DOI: 10.1093/nar/20.16.4187. View

16.

Shimizu S, Kamiike W, Hatanaka N, Nishimura M, Miyata M, Inoue T . Enzyme release from mitochondria during reoxygenation of rat liver. Transplantation. 1994; 57(1):144-8. DOI: 10.1097/00007890-199401000-00022. View

17.

Pfeiffer D, Gudz T, Novgorodov S, Erdahl W . The peptide mastoparan is a potent facilitator of the mitochondrial permeability transition. J Biol Chem. 1995; 270(9):4923-32. DOI: 10.1074/jbc.270.9.4923. View

18.

Jacobson M, Burne J, King M, Miyashita T, Reed J, Raff M . Bcl-2 blocks apoptosis in cells lacking mitochondrial DNA. Nature. 1993; 361(6410):365-9. DOI: 10.1038/361365a0. View

19.

Bernardi P, Vassanelli S, Veronese P, Colonna R, Szabo I, Zoratti M . Modulation of the mitochondrial permeability transition pore. Effect of protons and divalent cations. J Biol Chem. 1992; 267(5):2934-9. View

20.

Farrow S, White J, Martinou I, Raven T, Pun K, Grinham C . Cloning of a bcl-2 homologue by interaction with adenovirus E1B 19K. Nature. 1995; 374(6524):731-3. DOI: 10.1038/374731a0. View