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Acetic acid triggers cytochrome c release in yeast heterologously expressing human Bax

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Abstract

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.

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Funding

This work was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of “Contrato-Programa” UIDB/04050/2020- PE 20-23 and FCTANR/BEX-BCM/0175/2012. Joana P. Guedes acknowledges the PhD fellowship SFRH/BD/132070/2017 funded by FCT. Cátia Santos-Pereira acknowledges the PhD fellowship PD/BD/128032/2016 funded by FCT under the scope of the doctoral programme in Applied and Environmental Microbiology (DP_AEM).

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Author notes

  1. Joana P. Guedes and Vitória Baptista contributed equally to this study and share the first authorship.

Authors and Affiliations

  1. Centre of Molecular and Environmental Biology (CBMA), Department of Biology, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal

    Joana P. Guedes, Vitória Baptista, Cátia Santos-Pereira, Maria João Sousa, Susana R. Chaves & Manuela Côrte-Real

  2. Centro de Investigacíon Médica Aplicada (CIMA), Universidad de Navarra, 31008, Pamplona, Spain

    Joana P. Guedes

  3. Microelectromechanical Systems Research Unit (CMEMS-UMinho), School of Engineering, University of Minho, Campus de Azurém, 4800-058, Guimarães, Portugal

    Vitória Baptista

  4. Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal

    Vitória Baptista

  5. ICVS/3B’s – PT Government Associate Laboratory, Braga/Guimarães, Portugal

    Vitória Baptista

  6. Centre of Biological Engineering (CEB), Department of Biological Engineering, University of Minho, 4710-057, Braga, Portugal

    Cátia Santos-Pereira

  7. LABBELS – Associate Laboratory, Braga, Guimarães, Portugal

    Cátia Santos-Pereira

  8. UMR 5095, CNRS, Université de Bordeaux, Campus Carreire, 1 Rue Camille Saint-Saëns, 33077, Bordeaux, France

    Stéphen Manon

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Contributions

SRC, SM and MCR conceived and designed the experiments and analyzed the data; the first draft of the manuscript was written by JPG and VB and all authors commented on previous versions of the manuscript; material preparation, data collection and analysis were performed by JPG, VB, CSP and SM; SRC and MCR supervised the work, and MCR, MJS and SM contributed with reagents, materials and resources. All authors read and approved the final submitted version of the manuscript.

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Correspondence to Manuela Côrte-Real.

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Guedes, J.P., Baptista, V., Santos-Pereira, C. et al. Acetic acid triggers cytochrome c release in yeast heterologously expressing human Bax. Apoptosis 27, 368–381 (2022). https://doi.org/10.1007/s10495-022-01717-0

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