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Complete activation of Bax by a single site mutation

Oncogene volume 26, pages 7092–7102 (2007)Cite this article

Abstract

Bax translocation from the cytosol to mitochondria culminates a key step by which this protein mediates cell death. Here, we identified two amino acids, L70 and D71, within the BH3 domain of Bax that play a critical role in regulating Bax's cytosolic vs mitochondrial distribution. Individual substitution of these amino acids with alanine resulted in Bax conformational change, oligomerization, localization to mitochondria and cell death. Further mutational analysis indicated that L70 interacts with T174, V177 and A178 of Bax's C-terminal hydrophobic segment, while the negative charge of D71 is required for maintaining Bax in its soluble monomeric state. In summary, we have identified a new regulatory site that controls Bax's subcellular distribution and activation.

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References

  • Adams JM, Cory S . (1998). The Bcl-2 protein family: arbiters of cell survival. Science 281: 1322–1326.

    Article  CAS  Google Scholar 

  • Bowie JU, Luthy R, Eisenberg D . (1991). A method to identify protein sequences that fold into a known three-dimensional structure. Science 253: 164–170.

    Article  CAS  Google Scholar 

  • Cartron PF, Priault M, Oliver L, Meflah K, Manon S, Vallette FM . (2003). The N-terminal end of Bax contains a mitochondrial-targeting signal. J Biol Chem 278: 11633–11641.

    Article  CAS  Google Scholar 

  • Desagher S, Osen-Sand A, Nichols A, Eskes R, Montessuit S, Lauper S et al. (1999). Bid-induced conformational change of Bax is responsible for mitochondrial cytochrome c release during apoptosis. J Cell Biol 144: 891–901.

    Article  CAS  Google Scholar 

  • Eskes R, Desagher S, Antonsson B, Martinou JC . (2000). Bid induces the oligomerization and insertion of Bax into the outer mitochondrial membrane. Mol Cell Biol 20: 929–935.

    Article  CAS  Google Scholar 

  • Finucane DM, Bossy-Wetzel E, Waterhouse NJ, Cotter TG, Green DR . (1999). Bax-induced caspase activation and apoptosis via cytochrome c release from mitochondria is inhibitable by Bcl-XL. J Biol Chem 274: 2225–2233.

    Article  CAS  Google Scholar 

  • Goping IS, Gross A, Lavoie JN, Nguyen M, Jemmerson R, Roth K et al. (1998). Regulated targeting of Bax to mitochondria. J Cell Biol 143: 207–215.

    Article  CAS  Google Scholar 

  • Gross A, Jockel J, Wei MC, Korsmeyer SJ . (1998). Enforced dimerization of Bax results in its translocation, mitochondrial dysfunction and apoptosis. EMBO J 17: 3878–3885.

    Article  CAS  Google Scholar 

  • Hsu Y-T, Wolter KG, Youle RJ . (1997). Cytosol-to-membrane redistribution of Bax and Bcl-XL during apoptosis. Pro Natl Acad Sci USA 94: 3668–3672.

    Article  CAS  Google Scholar 

  • Hsu Y-T, Youle RJ . (1997). Nonionic detergents induce dimerization among members of the Bcl-2 family. J Biol Chem 272: 13829–13834.

    Article  CAS  Google Scholar 

  • Hsu Y-T, Youle RJ . (1998). Bax in murine thymus is a soluble monomeric protein that displays differential detergent-induced conformations. J Biol Chem 273: 10777–10783.

    Article  CAS  Google Scholar 

  • Kuwana T, Bouchier-Hayes L, Chipuk JE, Bonzon C, Sullivan BA, Green DR et al. (2005). BH3 domains of BH3-only proteins differentially regulate Bax-mediated mitochondrial membrane permeabilization both directly and indirectly. Mol Cell 17: 525–535.

    Article  CAS  Google Scholar 

  • Li P, Nijhawan D, Budihardjo I, Srinivasula SM, Ahmad M, Alnemri ES et al. (1997). Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 91: 479–489.

    Article  CAS  Google Scholar 

  • Manon S, Chaudhuri B, Guerin M . (1997). Release of cytochrome c and decrease of cytochrome c oxidase in Bax-expressing yeast cells, and prevention of these effects by coexpression of Bcl-XL. FEBS Lett 415: 29–32.

    Article  CAS  Google Scholar 

  • Mateu MG, Fersht AR . (1999). Mutually compensatory mutations during evolution of the tetramerization domain of tumor suppressor p53 lead to impaired hetero-oligomerization. Proc Natl Acad Sci USA 96: 3595–3599.

    Article  CAS  Google Scholar 

  • Meijerink JP, Mensink EJ, Wang K, Sedlak TW, Sloetjes AW, Witte TD et al. (1998). Hematopoietic malignancies demonstrate loss-of-function mutations of BAX. Blood 91: 2991–2997.

    CAS  PubMed  Google Scholar 

  • Mikhailov V, Mikhailova M, Pulkrabek DJ, Dong Z, Venkatachalam MA, Saikumar P . (2001). Bcl-2 prevents Bax oligomerization in the mitochondrial outer membrane. J Biol Chem 276: 18361–18374.

    Article  CAS  Google Scholar 

  • Nechushtan A, Smith CL, Hsu Y-T, Youle RJ . (1999). Conformation of the Bax C-terminus regulates subcellular location and cell death. EMBO J 18: 2330–2341.

    Article  CAS  Google Scholar 

  • Nechushtan A, Smith CL, Lamensdorf I, Yoon SH, Youle RJ . (2001). Bax and Bak coalesce into novel mitochondria-associated clusters during apoptosis. J Cell Biol 153: 1265–1276.

    Article  CAS  Google Scholar 

  • Oltvai ZN, Milliman CL, Korsmeyer SJ . (1993). Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74: 609–619.

    Article  CAS  Google Scholar 

  • Pagliari LJ, Kuwana T, Bonzon C, Newmeyer DD, Tu S, Beere HM et al. (2005). The multidomain proapoptotic molecules Bax and Bak are directly activated by heat. Proc Natl Acad Sci USA 102: 17975–17980.

    Article  CAS  Google Scholar 

  • Rampino N, Yamamoto H, Ionov Y, Li Y, Sawai H, Reed JC et al. (1997). Somatic frameshift mutations in the BAX gene in colon cancers of the microsatellite mutator phenotype. Science 275: 967–969.

    Article  CAS  Google Scholar 

  • Rosse T, Olivier R, Monney L, Rager M, Conus S, Fellay I et al. (1998). Bcl-2 prolongs cell survival after Bax-induced release of cytochrome c. Nature 391: 496–499.

    Article  CAS  Google Scholar 

  • Smaili SS, Hsu Y-T, Sanders KM, Russell JT, Youle RJ . (2001). Bax translocation to mitochondria subsequent to a rapid loss of mitochondrial membrane potential. Cell Death Differ 8: 909–920.

    Article  CAS  Google Scholar 

  • Suzuki M, Youle RJ, Tjandra N . (2000). Structure of Bax. Coregulation of dimer formation and intracellular localization. Cell 103: 645–654.

    Article  CAS  Google Scholar 

  • Thompson CB . (1995). Apoptosis in the pathogenesis and treatment of disease. Science 267: 1456–1462.

    Article  CAS  Google Scholar 

  • Wang K, Gross A, Waksman G, Korsmeyer SJ . (1998). Mutagenesis of the BH3 domain of BAX identifies residues critical for dimerization and killing. Mol Cell Biol 18: 6083–6089.

    Article  CAS  Google Scholar 

  • Willis SN, Chen L, Dewson G, Wei A, Naik E, Fletcher JI et al. (2005). Proapoptotic Bak is sequestered by Mcl-1 and Bcl-xL, but not Bcl-2, until displaced by BH3-only proteins. Genes Dev 19: 1294–1305.

    Article  CAS  Google Scholar 

  • Wolter KG, Hsu Y-T, Smith CL, Nechushtan A, Xi X-G, Youle RJ . (1997). Movement of Bax from the cytosol to mitochondria during apoptosis. J Cell Biol 139: 1281–1292.

    Article  CAS  Google Scholar 

  • Yin X-M, Oltvai ZN, Korsmeyer SJ . (1994). BH1 and BH2 domains of Bcl-2 are required for inhibition of apoptosis and heterodimerization with Bax. Nature 369: 321–323.

    Article  CAS  Google Scholar 

  • Zha H, Aime-Sempe C, Sato T, Reed JC . (1996). Proapoptotic protein Bax heterodimerizes with Bcl-2 and homodimerizes with Bax via a novel domain (BH3) distinct from BH1 and BH2. J Biol Chem 271: 7440–7444.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr Debbie Hansen for critical reading and Yunfei Chai for technical assistance. This research was supported by the grants RO1 NS40932 and COG RR01 5455 (MUSC animal facility) from the NIH.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC, USA

    H Zhou, Q Hou, J L Hansen & Y-T Hsu

  2. Department of Pharmacology, Institute of Materia Medica, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, PR China

    Q Hou

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  1. H Zhou
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  2. Q Hou
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  3. J L Hansen
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  4. Y-T Hsu
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Correspondence to Y-T Hsu.

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Supplementary information accompanies the paper on the Oncogene web site (http://www.nature.com/onc).

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Zhou, H., Hou, Q., Hansen, J. et al. Complete activation of Bax by a single site mutation. Oncogene 26, 7092–7102 (2007). https://doi.org/10.1038/sj.onc.1210517

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