Acetic acid triggers cytochrome c release in yeast heterologously expressing human Bax.
Acetic acid
Apoptosis
Bax
Bcl-2 family proteins
Heterologous expression
Yeast
Journal
Apoptosis : an international journal on programmed cell death
ISSN: 1573-675X
Titre abrégé: Apoptosis
Pays: Netherlands
ID NLM: 9712129
Informations de publication
Date de publication:
06 2022
06 2022
Historique:
accepted:
12
02
2022
pubmed:
2
4
2022
medline:
28
5
2022
entrez:
1
4
2022
Statut:
ppublish
Résumé
Proteins of the Bcl-2 protein family, including pro-apoptotic Bax and anti-apoptotic Bcl-xL, are critical for mitochondrial-mediated apoptosis regulation. Since yeast lacks obvious orthologs of Bcl-2 family members, heterologous expression of these proteins has been used to investigate their molecular and functional aspects. Active Bax is involved in the formation of mitochondrial outer membrane pores, through which cytochrome c (cyt c) is released, triggering a cascade of downstream apoptotic events. However, when in its inactive form, Bax is largely cytosolic or weakly bound to mitochondria. Given the central role of Bax in apoptosis, studies aiming to understand its regulation are of paramount importance towards its exploitation as a therapeutic target. So far, studies taking advantage of heterologous expression of human Bax in yeast to unveil regulation of Bax activation have relied on the use of artificial mutated or mitochondrial tagged Bax for its activation, rather than the wild type Bax (Bax α). Here, we found that cell death could be triggered in yeast cells heterologoulsy expressing Bax α with concentrations of acetic acid that are not lethal to wild type cells. This was associated with Bax mitochondrial translocation and cyt c release, closely resembling the natural Bax function in the cellular context. This regulated cell death process was reverted by co-expression with Bcl-xL, but not with Bcl-xLΔC, and in the absence of Rim11p, the yeast ortholog of mammalian GSK3β. This novel system mimics human Bax α regulation by GSK3β and can therefore be used as a platform to uncover novel Bax regulators and explore its therapeutic modulation.
Identifiants
pubmed: 35362903
doi: 10.1007/s10495-022-01717-0
pii: 10.1007/s10495-022-01717-0
doi:
Substances chimiques
Carrier Proteins
0
Proto-Oncogene Proteins c-bcl-2
0
bcl-2-Associated X Protein
0
bcl-X Protein
0
Cytochromes c
9007-43-6
Glycogen Synthase Kinase 3 beta
EC 2.7.11.1
Acetic Acid
Q40Q9N063P
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
368-381Informations de copyright
© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.
Références
Ashkenazi A, Salvesen G (2014) Regulated cell death: signaling and mechanisms. Annu Rev Cell Dev Biol 30:337–356. https://doi.org/10.1146/annurev-cellbio-100913-013226
doi: 10.1146/annurev-cellbio-100913-013226
pubmed: 25150011
Elmore S (2007) Apoptosis: a review of programmed cell death. Toxicol Pathol 35:495–516. https://doi.org/10.1080/01926230701320337
doi: 10.1080/01926230701320337
pubmed: 17562483
pmcid: 2117903
Thompson CB (1995) Apoptosis in the pathogenesis and treatment of disease. Science (80-). https://doi.org/10.1126/science.7878464
Chipuk JE, Moldoveanu T, Llambi F et al (2010) The BCL-2 family reunion. Mol Cell 37:299–310. https://doi.org/10.1016/j.molcel.2010.01.025
doi: 10.1016/j.molcel.2010.01.025
pubmed: 20159550
pmcid: 3222298
Adams JM, Cory S (2007) The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene 26:1324–1337. https://doi.org/10.1038/sj.onc.1210220
doi: 10.1038/sj.onc.1210220
pubmed: 17322918
pmcid: 2930981
Adams JM, Cory S (1998) The Bcl-2 protein family: arbiters of cell survival. Science 281:1322–1326. https://doi.org/10.1126/science.281.5381.1322
doi: 10.1126/science.281.5381.1322
pubmed: 9735050
Kale J, Osterlund EJ, Andrews DW (2018) BCL-2 family proteins: changing partners in the dance towards death. Cell Death Differ 25:65–80. https://doi.org/10.1038/cdd.2017.186
doi: 10.1038/cdd.2017.186
pubmed: 29149100
Peña-Blanco A, García-Sáez AJ (2018) Bax, Bak and beyond - mitochondrial performance in apoptosis. FEBS J 285:416–431. https://doi.org/10.1111/febs.14186
doi: 10.1111/febs.14186
pubmed: 28755482
Te HY, Youle RJ (1998) Bax in murine thymus is a soluble monomeric protein that displays differential detergent-induced conformations. J Biol Chem. https://doi.org/10.1074/jbc.273.17.10777
doi: 10.1074/jbc.273.17.10777
Te HY, Wolter KG, Youle RJ (1997) Cytosol-to-membrane redistribution of Bax and Bcl-XL during apoptosis. Proc Natl Acad Sci U S A. https://doi.org/10.1073/pnas.94.8.3668
doi: 10.1073/pnas.94.8.3668
Zong W-X, Li C, Hatzivassiliou G et al (2003) Bax and Bak can localize to the endoplasmic reticulum to initiate apoptosis. J Cell Biol 162:59–69. https://doi.org/10.1083/jcb.200302084
doi: 10.1083/jcb.200302084
pubmed: 12847083
pmcid: 2172724
Renault TT, Manon S (2011) Bax: addressed to kill. Biochimie 93:1379–1391. https://doi.org/10.1016/j.biochi.2011.05.013
doi: 10.1016/j.biochi.2011.05.013
pubmed: 21641962
Wolter KG, Te HY, Smith CL et al (1997) Movement of Bax from the cytosol to mitochondria during apoptosis. J Cell Biol. https://doi.org/10.1083/jcb.139.5.1281
doi: 10.1083/jcb.139.5.1281
pubmed: 9382873
pmcid: 2140220
Lalier L, Cartron P-F, Juin P et al (2007) Bax activation and mitochondrial insertion during apoptosis. Apoptosis 12:887–896. https://doi.org/10.1007/s10495-007-0749-1
doi: 10.1007/s10495-007-0749-1
pubmed: 17453158
Kirchhoff SR, Gupta S, Knowlton AA (2002) Cytosolic heat shock protein 60, apoptosis, and myocardial injury. Circulation 105:2899–2904. https://doi.org/10.1161/01.cir.0000019403.35847.23
doi: 10.1161/01.cir.0000019403.35847.23
pubmed: 12070120
Zong WX, Lindsten T, Ross AJ et al (2001) BH3-only proteins that bind pro-survival Bcl-2 family members fail to induce apoptosis in the absence of Bax and Bak. Genes Dev. https://doi.org/10.1101/gad.897601
doi: 10.1101/gad.897601
pubmed: 11410528
pmcid: 312722
Kuwana T, Bouchier-Hayes L, Chipuk JE 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. https://doi.org/10.1016/j.molcel.2005.02.003
doi: 10.1016/j.molcel.2005.02.003
pubmed: 15721256
Gardai SJ, Hildeman DA, Frankel SK et al (2004) Phosphorylation of Bax ser184 by Akt regulates its activity and apoptosis in neutrophils. J Biol Chem. https://doi.org/10.1074/jbc.M400063200
doi: 10.1074/jbc.M400063200
pubmed: 15292176
Xin M, Gao F, May WS et al (2007) Protein kinase Cζ abrogates the proapoptotic function of bax through phosphorylation. J Biol Chem. https://doi.org/10.1074/jbc.M701613200
doi: 10.1074/jbc.M701613200
pubmed: 18089575
Xin M, Deng X (2006) Protein phosphatase 2A enhances the proapoptotic function of Bax through dephosphorylation. J Biol Chem 281:18859–18867. https://doi.org/10.1074/jbc.M512543200
doi: 10.1074/jbc.M512543200
pubmed: 16679323
Linseman DA, Butts BD, Precht TA et al (2004) Glycogen synthase kinase-3β phosphorylates Bax and promotes its mitochondrial localization during neuronal apoptosis. J Neurosci 24:9993–10002. https://doi.org/10.1523/JNEUROSCI.2057-04.2004
doi: 10.1523/JNEUROSCI.2057-04.2004
pubmed: 15525785
pmcid: 6730230
Kim BJ, Ryu SW, Song BJ (2006) JNK- and p38 kinase-mediated phosphorylation of Bax leads to its activation and mitochondrial translocation and to apoptosis of human hepatoma HepG2 cells. J Biol Chem. https://doi.org/10.1074/jbc.M510644200
doi: 10.1074/jbc.M510644200
pubmed: 17197697
Carmona-Gutierrez D, Bauer MA, Zimmermann A, et al (2018) Guidelines and recommendations on yeast cell death nomenclature. Microb Cell (Graz, Austria) 5:4–31. https://doi.org/10.15698/mic2018.01.607
Carmona-Gutierrez D, Eisenberg T, Büttner S et al (2010) Apoptosis in yeast: triggers, pathways, subroutines. Cell Death Differ 17:763–773. https://doi.org/10.1038/cdd.2009.219
doi: 10.1038/cdd.2009.219
pubmed: 20075938
Priault M, Camougrand N, Kinnally KW et al (2003) Yeast as a tool to study Bax/mitochondrial interactions in cell death. FEMS Yeast Res 4:15–27. https://doi.org/10.1016/S1567-1356(03)00143-0
doi: 10.1016/S1567-1356(03)00143-0
pubmed: 14554193
Pereira C, Silva RD, Saraiva L et al (2008) Mitochondria-dependent apoptosis in yeast. Biochim Biophys Acta 1783:1286–1302. https://doi.org/10.1016/j.bbamcr.2008.03.010
doi: 10.1016/j.bbamcr.2008.03.010
pubmed: 18406358
Polčic P, Jaká P, Mentel M (2015) Yeast as a tool for studying proteins of the Bcl-2 family. Microb Cell (Graz, Austria) 2:74–87. https://doi.org/10.15698/mic2015.03.193
Alves S, Neiri L, Chaves SR et al (2018) N-terminal acetylation modulates Bax targeting to mitochondria. Int J Biochem Cell Biol 95:35–42. https://doi.org/10.1016/j.biocel.2017.12.004
doi: 10.1016/j.biocel.2017.12.004
pubmed: 29233735
Arokium H, Ouerfelli H, Velours G et al (2007) Substitutions of potentially phosphorylatable serine residues of bax reveal how they may regulate its interaction with mitochondria. J Biol Chem. https://doi.org/10.1074/jbc.M704891200
doi: 10.1074/jbc.M704891200
pubmed: 17911107
Renault TT, Teijido O, Missire F et al (2015) Bcl-xL stimulates Bax relocation to mitochondria and primes cells to ABT-737. Int J Biochem Cell Biol 64:136–146. https://doi.org/10.1016/j.biocel.2015.03.020
doi: 10.1016/j.biocel.2015.03.020
pubmed: 25862283
Gietz RD, Akio S (1988) New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene 74:527–534. https://doi.org/10.1016/0378-1119(88)90185-0
doi: 10.1016/0378-1119(88)90185-0
pubmed: 3073106
Garenne D, Renault TT, Manon S (2016) Bax mitochondrial relocation is linked to its phosphorylation and its interaction with Bcl-xL. Microb Cell (Graz, Austria) 3:597–605. https://doi.org/10.15698/mic2016.12.547
Millard PJ, Roth BL, Thi HPT et al (1997) Development of the FUN-1 family of fluorescent probes for vacuole labeling and viability testing of yeasts. Appl Environ Microbiol. https://doi.org/10.1128/aem.63.7.2897-2905.1997
doi: 10.1128/aem.63.7.2897-2905.1997
pubmed: 9212436
pmcid: 168585
Prudêncio C, Sansonetty F, Côrte-Real M (1998) Flow cytometric assessment of cell structural and functional changes induced by acetic acid in the yeasts Zygosaccharomyces bailii and Saccharomyces cerevisiae. Cytometry. https://doi.org/10.1002/(SICI)1097-0320(19980401)31:4%3c307::AID-CYTO11%3e3.0.CO;2-U
doi: 10.1002/(SICI)1097-0320(19980401)31:4<307::AID-CYTO11>3.0.CO;2-U
pubmed: 9551607
Camougrand N, Grelaud-Coq A, Marza E et al (2003) The product of the UTH1 gene, required for Bax-induced cell death in yeast, is involved in the response to rapamycin. Mol Microbiol. https://doi.org/10.1046/j.1365-2958.2003.03311.x
doi: 10.1046/j.1365-2958.2003.03311.x
pubmed: 12519199
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
doi: 10.1016/S0021-9258(19)52451-6
pubmed: 14907713
Westphal D, Kluck RM, Dewson G (2014) Building blocks of the apoptotic pore: how Bax and Bak are activated and oligomerize during apoptosis. Cell Death Differ 21:196–205. https://doi.org/10.1038/cdd.2013.139
doi: 10.1038/cdd.2013.139
pubmed: 24162660
Schellenberg B, Wang P, Keeble JA et al (2013) Bax exists in a dynamic equilibrium between the cytosol and mitochondria to control apoptotic priming. Mol Cell 49:959–971. https://doi.org/10.1016/j.molcel.2012.12.022
doi: 10.1016/j.molcel.2012.12.022
pubmed: 23375500
pmcid: 3594749
Pereira H, Oliveira CSF, Castro L, et al (2015) Yeast as a tool to explore cathepsin D function. Microb Cell (Graz, Austria) 2:225–234. https://doi.org/10.15698/mic2015.07.212
Kroemer G, Galluzzi L, Vandenabeele P et al (2009) Classification of cell death: recommendations of the Nomenclature Committee on Cell Death 2009. Cell Death Differ 16:3–11. https://doi.org/10.1038/cdd.2008.150
doi: 10.1038/cdd.2008.150
pubmed: 18846107
Renault TT, Dejean LM, Manon S (2017) A brewing understanding of the regulation of Bax function by Bcl-xL and Bcl-2. Mech Ageing Dev 161:201–210. https://doi.org/10.1016/j.mad.2016.04.007
doi: 10.1016/j.mad.2016.04.007
pubmed: 27112371
Simonyan L, Renault TT, da Costa Novais MJ et al (2016) Regulation of Bax/mitochondria interaction by AKT. FEBS Lett 590:13–21. https://doi.org/10.1002/1873-3468.12030
doi: 10.1002/1873-3468.12030
pubmed: 26763134
Gallenne T, Gautier F, Oliver L et al (2009) Bax activation by the BH3-only protein Puma promotes cell dependence on antiapoptotic Bcl-2 family members. J Cell Biol 185:279–290. https://doi.org/10.1083/jcb.200809153
doi: 10.1083/jcb.200809153
pubmed: 19380879
pmcid: 2700382
Greenhalf W, Stephan C, Chaudhuri B (1996) Role of mitochondria and C-terminal membrane anchor of Bcl-2 in Bax induced growth arrest and mortality in Saccharomyces cerevisiae. FEBS Lett 380:169–175. https://doi.org/10.1016/0014-5793(96)00044-0
doi: 10.1016/0014-5793(96)00044-0
pubmed: 8603730
Manon S, Chaudhuri B, Guérin M (1997) Release of cytochrome c and decrease of cytochrome c oxidase in Bar-expressing yeast cells, and prevention of these effects by coexpression of Bcl-x(L). FEBS Lett. https://doi.org/10.1016/S0014-5793(97)01087-9
doi: 10.1016/S0014-5793(97)01087-9
pubmed: 9326363
Torgler CN, de Tiani M, Raven T et al (1997) Expression of bak in S. pombe results in a lethality mediated through interaction with the calnexin homologue Cnx1. Cell Death Differ 4:263–271. https://doi.org/10.1038/sj.cdd.4400239
doi: 10.1038/sj.cdd.4400239
pubmed: 16465239
Bounhar Y, Mann KK, Roucou X, LeBlanc AC (2006) Prion protein prevents Bax-mediated cell death in the absence of other Bcl-2 family members in Saccharomyces cerevisiae. FEMS Yeast Res 6:1204–1212. https://doi.org/10.1111/j.1567-1364.2006.00122.x
doi: 10.1111/j.1567-1364.2006.00122.x
pubmed: 17156017
Ligr M, Madeo F, Fröhlich E et al (1998) Mammalian Bax triggers apoptotic changes in yeast. FEBS Lett 438:61–65. https://doi.org/10.1016/s0014-5793(98)01227-7
doi: 10.1016/s0014-5793(98)01227-7
pubmed: 9821959
De Smet K, Eberhardt I, Contreras R (2004) Bax-induced cell death in Candida albicans. Yeast. https://doi.org/10.1002/yea.1180
doi: 10.1002/yea.1180
pubmed: 15565645
Poliaková D, Sokolíková B, Kolarov J, Šabová L (2002) The antiapoptotic protein Bcl-XL prevents the cytotoxic effect of Bax, but not Bax-induced formation of reactive oxygen species, in Kluyveromyces lactis. Microbiology. https://doi.org/10.1099/00221287-148-9-2789
doi: 10.1099/00221287-148-9-2789
pubmed: 12449634
Arokium H, Camougrand N, Vallette FM, Manon S (2004) Studies of the Interaction of Substituted Mutants of BAX with Yeast Mitochondria Reveal That the C-terminal Hydrophobic α-Helix Is a Second ART Sequence and Plays a Role in the Interaction with Anti-apoptotic BCL-x
doi: 10.1074/jbc.M408373200
pubmed: 15459197
Cartron P-F, Arokium H, Oliver L et al (2005) Distinct domains control the addressing and the insertion of Bax into mitochondria. J Biol Chem 280:10587–10598. https://doi.org/10.1074/jbc.M409714200
doi: 10.1074/jbc.M409714200
pubmed: 15590655
Casey E, Sedlak M, Ho NWY, Mosier NS (2010) Effect of acetic acid and pH on the cofermentation of glucose and xylose to ethanol by a genetically engineered strain of Saccharomyces cerevisiae. FEMS Yeast Res 10:385–393. https://doi.org/10.1111/j.1567-1364.2010.00623.x
doi: 10.1111/j.1567-1364.2010.00623.x
pubmed: 20402796
Casal M, Cardoso H, Leao C (1996) Mechanisms regulating the transport of acetic acid in Saccharomyces cerevisiae. Microbiology 142:1385–1390. https://doi.org/10.1099/13500872-142-6-1385
doi: 10.1099/13500872-142-6-1385
pubmed: 8704978
Jan G, Belzacq A-S, Haouzi D et al (2002) Propionibacteria induce apoptosis of colorectal carcinoma cells via short-chain fatty acids acting on mitochondria. Cell Death Differ 9:179–188. https://doi.org/10.1038/sj.cdd.4400935
doi: 10.1038/sj.cdd.4400935
pubmed: 11840168
Lan A, Lagadic-Gossmann D, Lemaire C et al (2007) Acidic extracellular pH shifts colorectal cancer cell death from apoptosis to necrosis upon exposure to propionate and acetate, major end-products of the human probiotic propionibacteria. Apoptosis 12:573–591. https://doi.org/10.1007/s10495-006-0010-3
doi: 10.1007/s10495-006-0010-3
pubmed: 17195096
Marques C, Oliveira CSF, Alves S et al (2013) Acetate-induced apoptosis in colorectal carcinoma cells involves lysosomal membrane permeabilization and cathepsin D release. Cell Death Dis 4:e507–e507. https://doi.org/10.1038/cddis.2013.29
doi: 10.1038/cddis.2013.29
pubmed: 23429293
pmcid: 3734821
Saraiva L, Silva RD, Pereira G et al (2006) Specific modulation of apoptosis and Bcl-xL phosphorylation in yeast by distinct mammalian protein kinase C isoforms. J Cell Sci 119:3171–3181. https://doi.org/10.1242/jcs.03033
doi: 10.1242/jcs.03033
pubmed: 16835272
Edlich F, Banerjee S, Suzuki M et al (2011) Bcl-x(L) retrotranslocates Bax from the mitochondria into the cytosol. Cell 145:104–116. https://doi.org/10.1016/j.cell.2011.02.034
doi: 10.1016/j.cell.2011.02.034
pubmed: 21458670
pmcid: 3070914
Todt F, Cakir Z, Reichenbach F et al (2013) The C-terminal helix of Bcl-x L mediates Bax retrotranslocation from the mitochondria. Cell Death Differ. https://doi.org/10.1038/cdd.2012.131
doi: 10.1038/cdd.2012.131
pubmed: 23079612
Zhang L, Zhang Y, Xing D (2010) LPLI inhibits apoptosis upstream of Bax translocation via a GSK-3β-inactivation mechanism. J Cell Physiol. https://doi.org/10.1002/jcp.22123
doi: 10.1002/jcp.22123
pubmed: 20589837
pmcid: 2971691