Bcl-xL stimulates Bax relocation to mitochondria and primes cells to ABT-737

Renault, Thibaud T.; Teijido, Oscar; Missire, Florent; Ganesan, Yogesh Tengarai; Velours, Gisèle; Arokium, Hubert; Beaumatin, Florian; Llanos, Raul; Athané, A.; Camougrand, Nadine; Priault, Muriel; Antonsson, Bruno; Dejean, Laurent M.; Manon, Stéphen

doi:10.1016/j.biocel.2015.03.020

Cited by 36 publications

(63 citation statements)

References 29 publications

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“…Like previously reported for BaxWT 24, the co-expression of full-length Bcl-xL stimulated the mitochondrial localization of Bax-S184D (Fig. 1A, B).…”

Section: Resultssupporting

confidence: 86%

Section: Introductionsupporting

confidence: 61%

“…Furthermore, when a truncated mutant Bcl-xLΔC was co-expressed with Bax, the stimulation of Bax mitochondrial localization was even greater than wilth full-length Bcl-xL, both in mammalian cells and in yeast 24. Indeed, while Bcl-xL is able to retrotranslocate Bax from the mitochondria to the cytosol 25, Bcl-xLΔC lacks this ability 2426 and stimulates Bax activation 24. These experiments and others 27 converge to support a model of dynamic equilibrium of Bax localization, that would be controlled, namely, by the phosphorylation of Bax and the interaction with Bcl-xL.…”

Section: Introductionmentioning

confidence: 98%

“…Under non-apoptotic conditions, Bcl-xL-overexpression increases the mitochondrial localization of Bax in parental mouse prolymphocytic cells FL5.12 or in human colorectal cancer cells HCT-116 24. A similar observation was made in yeast, where the co-expression of Bax and Bcl-xL induced a greater mitochondrial localization of Bax than when it was expressed alone 24. This suggests that Bax translocation was related to intrinsic characteristics of the interaction between Bax and Bcl-xL, that were independent from the mammalian cellular context.…”

Section: Introductionmentioning

confidence: 99%

“…This suggests that Bax translocation was related to intrinsic characteristics of the interaction between Bax and Bcl-xL, that were independent from the mammalian cellular context. Furthermore, when a truncated mutant Bcl-xLΔC was co-expressed with Bax, the stimulation of Bax mitochondrial localization was even greater than wilth full-length Bcl-xL, both in mammalian cells and in yeast 24. Indeed, while Bcl-xL is able to retrotranslocate Bax from the mitochondria to the cytosol 25, Bcl-xLΔC lacks this ability 2426 and stimulates Bax activation 24.…”

Section: Introductionmentioning

confidence: 99%

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Bax mitochondrial relocation is linked to its phosphorylation and its interaction with Bcl-xL

Garenne¹,

Renault²,

Manon³

2016

Microb Cell

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The heterologous expression of Bax, and other Bcl-2 family members, in the yeast Saccharomyces cerevisiae, has proved to be a valuable reporter system to investigate the molecular mechanisms underlying their interaction with mitochondria. By combining the co-expression of Bax and Bcl-xL mutants with analyzes of their localization and interaction in mitochondria and post-mitochondrial supernatants, we showed that the ability of Bax and Bcl-xL to interact is dependent both on Bax phosphorylation - mimicked by a substitution S184D - and by Bax and Bcl-xL localization. This, and previous data, provide the molecular basis for a model of dynamic equilibrium for Bax localization and activation, regulated both by phosphorylation and Bcl-xL.

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“…Like previously reported for BaxWT 24, the co-expression of full-length Bcl-xL stimulated the mitochondrial localization of Bax-S184D (Fig. 1A, B).…”

Section: Resultssupporting

confidence: 86%

Section: Introductionsupporting

confidence: 61%

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Bax mitochondrial relocation is linked to its phosphorylation and its interaction with Bcl-xL

Garenne¹,

Renault²,

Manon³

2016

Microb Cell

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Regulation of Bax/mitochondria interaction by AKT

et al. 2015

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Edited by Barry HalliwellBax-dependent mitochondrial permeabilization during apoptosis is controlled by multiple factors, including the phosphorylation by the protein kinase AKT. We used the heterologous co-expression of human Bax and AKT1 in yeast to investigate how the kinase modulates the different steps underlying Bax activation. We found that AKT activated Bax and increased its cellular content. Both effects were dependent on Ser184, but a phosphorylation of this residue did not fully explain the effects of AKT. Additional experiments with mutants substituted on Ser184 suggested that the regulation of Bax dynamic equilibrium between the cytosol and mitochondria might be more tightly regulated by Bcl-xL when Bax is phosphorylated.

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Investigating BCL-2 Family Protein Interactions in Yeast

Manon¹

2018

Methods in Molecular Biology

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Among the different models used to study the biochemical properties and function of proteins of the Bcl-2 family, their heterologous expression in the yeast Saccharomyces cerevisiae may look out of place. However, when grown under adequate conditions, yeast cells have mitochondria that have similar properties as those of mammalian cells, and are able to be targeted by mammalian Bcl-2 family members. Yeast thus provides a neutral cellular background to study how proteins of the Bcl-2 family interact with mitochondria, alone or in couple (or more). Most studies done in our laboratory has been done on the pro-apoptotic protein Bax and the anti-apoptotic protein Bcl-xL, but yeast can bring useful information about every protein of the family, in terms of their capacity to interact and to regulate the permeabilization of the outer mitochondrial membrane. 1.IntroductionStructure/functions studies of Bcl-2 family members, like for any other proteins, can be done in whole cells or in recombinant models. For the later, one difficulty is to obtain proteins under a conformation as close to the native conformation as possible. This may be a mimitation for a protein such as the proapoptotic protein Bax: indeed, Bax exhibits characteristics resembling those of a membrane protein, namely because of the presence of its C-terminal hydrophobic α-helix. For long, investigators have produced the protein deprived from this helix (e.g. [1]) or have used detergents to maintain the protein in solution and unaggregated (e.g. [2]). However, those surfactants may have the same membranepermeabilizing effects as active Bax and/or can favor the active conformation of Bax, that maybe a limitation to study its regulation [3]. The heterologous expression of Bcl-2 family members in yeast has been used very early as a middleterm alternative between the expression in mammalian cells and recombinant models. The first direct demonstration that Bax was able to permeabilize mitochondria to cytochrome c has been done in yeast [4], one year before the same demonstration in mammalian cells [5]. Yeast does not have homologs of the Bcl-2 family members, even though a form of programmed cell death is present in yeast ([6] for a review). However, when expressed in yeast, Bcl-2 family members keep their ability to interact with mitochondria and, under adequate conditions, to permeabilize yeast outer miitochondrial membrane to cytochrome c, providing a model to investigate Bax activity ([7] for review). In this chapter, we describe methods that have been improved for 20 years, that allows to investigate the function of Bax when it is expressed in yeast. These methods have been applied to the study of different Bax mutants [2,[8][9][10][11][12], but can also been applied to the study of combination with other Bcl-2 family members, such as 13], tBid [14], Puma [15], or zebrafish protein . It can also be used to study the role of other Bax regulation candidates such as Tom22 [17,18] or AKT [11]. 2.MaterialsPrepare all reagents and buffer in ultrapure water (18 M...

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Bcl-xL stimulates Bax relocation to mitochondria and primes cells to ABT-737

Cited by 36 publications

References 29 publications

Bax mitochondrial relocation is linked to its phosphorylation and its interaction with Bcl-xL

Bax mitochondrial relocation is linked to its phosphorylation and its interaction with Bcl-xL

Regulation of Bax/mitochondria interaction by AKT

Investigating BCL-2 Family Protein Interactions in Yeast

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