Yeast Bxi1/Ybh3 mediates conserved mitophagy and apoptosis in yeast and mammalian cells: convergence in Bcl-2 family
-
Yuying Wang
, Zhiyuan Hu , Maojun Jiang , Yanxin Zhang , Linjie Yuan , Ziqian Wang , Ting Songund Zhichao Zhang
Abstract
The process of degrading unwanted or damaged mitochondria by autophagy, called mitophagy, is essential for mitochondrial quality control together with mitochondrial apoptosis. In mammalian cells, pan-Bcl-2 family members including conical Bcl-2 members and non-conical ones are involved in and govern the two processes. We have illustrated recently the BH3 receptor Hsp70 interacts with Bim to mediate both apoptosis and mitophagy. However, whether similar pathways exist in lower eukaryotes where conical Bcl-2 members are absent remained unclear. Here, a specific inhibitor of the Hsp70-Bim PPI, S1g-10 and its analogs were used as chemical tools to explore the role of yeast Bxi1/Ybh3 in regulating mitophagy and apoptosis. Using Om45-GFP processing assay, we illustrated that yeast Ybh3 mediates a ubiquitin-related mitophagy pathway in both yeast and mammalian cells through association with Hsp70, which is in the same manner with Bim. Moreover, by using Bax/Bak double knockout MEF cells, Ybh3 was identified to induce apoptosis through forming oligomerization to trigger mitochondrial outer membrane permeabilization (MOMP) like Bax. We not only illustrated a conserved ubiquitin-related mitophagy pathway in yeast but also revealed the multi-function of Ybh3 which combines the function of BH3-only protein and multi-domain Bax protein as one.
Funding source: Fundamental Research Funds for the Central University
Award Identifier / Grant number: 2022ZXYG46, DUT22YG106, DUT22YG223, DUT23YG125
Funding source: National Natural Science Foundation of China
Award Identifier / Grant number: 82073703, 82270186, 82273778
Acknowledgments
The authors thank Prof. Xinguang Liu (Guangdong Medical University) for the kind gift of yeast strains.
-
Research ethics: Not applicable.
-
Author contributions: Conceptualization, YW, TS, and ZZ; data curation, YW, TS, and ZZ; Yeast and Cell culture, Western blotting and co-immunoprecipitation, YW, ZH; fluorescence properties detection and apoptosis assay, YW and YL; chemical synthesis, MJ and ZW; size-exclusion chromatography analysis, YW and YZ; writing-original draft preparation, YW, TS and ZZ; Writing-review & editing, WY, TS and ZZ; funding acquisition, TS and ZZ. All the authors have read and agreed to the published version of the manuscript. All authors read and approved the final manuscript.
-
Competing interests: The authors state no conflict of interest.
-
Research funding: This research was supported by the National Natural Science Foundation of China (82270186, 82073703 and 82273778) and the Fundamental Research Funds for the Central University (DUT22YG223, DUT22YG106, DUT23YG125, and 2022ZXYG46).
-
Data availability: The raw data can be obtained on request from the corresponding author.
References
Buttner, S., Ruli, D., Vogtle, F.N., Galluzzi, L., Moitzi, B., Eisenberg, T., Kepp, O., Habernig, L., Carmona-Gutierrez, D., Rockenfeller, P., et al.. (2011). A yeast BH3-only protein mediates the mitochondrial pathway of apoptosis. EMBO J. 30: 2779–2792, https://doi.org/10.1038/emboj.2011.197.Suche in Google Scholar PubMed PubMed Central
Cebulski, J., Malouin, J., Pinches, N., Cascio, V., and Austriaco, N. (2011). Yeast Bax inhibitor, Bxi1p, is an ER-localized protein that links the unfolded protein response and programmed cell death in Saccharomyces cerevisiae. PLoS One 6: e20882, https://doi.org/10.1371/journal.pone.0020882.Suche in Google Scholar PubMed PubMed Central
Chae, H.J., Ke, N., Kim, H.R., Chen, S., Godzik, A., Dickman, M., and Reed, J.C. (2003). Evolutionarily conserved cytoprotection provided by Bax inhibitor-1 homologs from animals, plants, and yeast. Gene 32: 3101–3113, https://doi.org/10.1016/j.gene.2003.09.011.Suche in Google Scholar PubMed
Czabotar, P.E. and Garcia-Saez, A.J. (2023). Mechanisms of BCL-2 family proteins in mitochondrial apoptosis. Nat. Rev. Mol. Cell Biol. 24: 732–748, https://doi.org/10.1038/s41580-023-00629-4.Suche in Google Scholar PubMed
Galluzzi, L., Kepp, O., and Kroemer, G. (2012). Mitochondria: master regulators of danger signalling. Nat. Rev. Mol. Cell Biol. 13: 780–788, https://doi.org/10.1038/nrm3479.Suche in Google Scholar PubMed
Garofalo, T., Matarrese, P., Manganelli, V., Marconi, M., Tinari, A., Gambardella, L., Faggioni, A., Misasi, R., Sorice, M., and Malorni, W. (2016). Evidence for the involvement of lipid rafts localized at the ER-mitochondria associated membranes in autophagosome formation. Autophagy 12: 917–935, https://doi.org/10.1080/15548627.2016.1160971.Suche in Google Scholar PubMed PubMed Central
Gelmetti, V., De Rosa, P., Torosantucci, L., Marini, E.S., Romagnoli, A., Di Rienzo, M., Arena, G., Vignone, D., Fimia, G.M., and Valente, E.M. (2017). PINK1 and BECN1 relocalize at mitochondria-associated membranes during mitophagy and promote ER-mitochondria tethering and autophagosome formation. Autophagy 13: 654–669, https://doi.org/10.1080/15548627.2016.1277309.Suche in Google Scholar PubMed PubMed Central
Gil-Parrado, S., Fernandez-Montalvan, A., Assfalg-Machleidt, I., Popp, O., Bestvater, F., Holloschi, A., Knoch, T.A., Auerswald, E.A., Welsh, K., Reed, J.C., et al.. (2002). Ionomycin-activated calpain triggers apoptosis. A probable role for Bcl-2 family members. J. Biol. Chem. 277: 27217–27226, https://doi.org/10.1074/jbc.m202945200.Suche in Google Scholar PubMed
Guo, Z., Song, T., Wang, Z., Lin, D., Cao, K., Liu, P., Feng, Y., Zhang, X., Wang, P., Yin, F., et al.. (2020). The chaperone Hsp70 is a BH3 receptor activated by the pro-apoptotic Bim to stabilize anti-apoptotic clients. J. Biol. Chem. 295: 12900–12909, https://doi.org/10.1074/jbc.ra120.013364.Suche in Google Scholar
Hill, B.G., Benavides, G.A., Lancaster, J.J., Ballinger, S., Dell’Italia, L., Jianhua, Z., and Darley-Usmar, V.M. (2012). Integration of cellular bioenergetics with mitochondrial quality control and autophagy. Biol. Chem. 393: 1485–1512, https://doi.org/10.1515/hsz-2012-0198.Suche in Google Scholar PubMed PubMed Central
Kanki, T., Kang, D., and Klionsky, D.J. (2009). Monitoring mitophagy in yeast: the Om45-GFP processing assay. Autophagy 5: 1186–1189, https://doi.org/10.4161/auto.5.8.9854.Suche in Google Scholar PubMed PubMed Central
Kanki, T. and Klionsky, D.J. (2008). Mitophagy in yeast occurs through a selective mechanism. J. Biol. Chem. 283: 32386–32393, https://doi.org/10.1074/jbc.m802403200.Suche in Google Scholar PubMed PubMed Central
Kumar, R. and Reichert, A.S. (2021). Common principles and specific mechanisms of mitophagy from yeast to humans. Int. J. Mol. Sci. 22: 4363, https://doi.org/10.3390/ijms22094363.Suche in Google Scholar PubMed PubMed Central
Lebeaupin, C., Blanc, M., Vallee, D., Keller, H., and Bailly-Maitre, B. (2020). BAX inhibitor-1: between stress and survival. FEBS J. 287: 1722–1736, https://doi.org/10.1111/febs.15179.Suche in Google Scholar PubMed PubMed Central
Lemasters, J.J. (2005). Selective mitochondrial autophagy, or mitophagy, as a targeted defense against oxidative stress, mitochondrial dysfunction, and aging. Rejuvenation Res. 8: 3–5, https://doi.org/10.1089/rej.2005.8.3.Suche in Google Scholar PubMed
Lisbona, F., Rojas-Rivera, D., Thielen, P., Zamorano, S., Todd, D., Martinon, F., Glavic, A., Kress, C., Lin, J.H., Walter, P., et al.. (2009). BAX inhibitor-1 is a negative regulator of the ER stress sensor IRE1alpha. Mol. Cell 33: 679–691, https://doi.org/10.1016/j.molcel.2009.02.017.Suche in Google Scholar PubMed PubMed Central
Maes, H., Kuchnio, A., Peric, A., Moens, S., Nys, K., De Bock, K., Quaegebeur, A., Schoors, S., Georgiadou, M., Wouters, J., et al.. (2014). Tumor vessel normalization by chloroquine independent of autophagy. Cancer Cell 26: 190–206, https://doi.org/10.1016/j.ccr.2014.06.025.Suche in Google Scholar PubMed
Mentel, M., Illova, M., Krajcovicova, V., Kroupova, G., Mannova, Z., Chovancikova, P., and Polcic, P. (2023). Yeast Bax inhibitor (Bxi1p/Ybh3p) is not required for the action of Bcl-2 family proteins on cell viability. Int. J. Mol. Sci. 24: 12011, https://doi.org/10.3390/ijms241512011.Suche in Google Scholar PubMed PubMed Central
Muller, M., Kotter, P., Behrendt, C., Walter, E., Scheckhuber, C.Q., Entian, K.D., and Reichert, A.S. (2015). Synthetic quantitative array technology identifies the Ubp3-Bre5 deubiquitinase complex as a negative regulator of mitophagy. Cell Rep. 10: 1215–1225, https://doi.org/10.1016/j.celrep.2015.01.044.Suche in Google Scholar PubMed
Okamoto, K., Kondo-Okamoto, N., and Ohsumi, Y. (2009). Mitochondria-anchored receptor Atg32 mediates degradation of mitochondria via selective autophagy. Dev. Cell 17: 87–97, https://doi.org/10.1016/j.devcel.2009.06.013.Suche in Google Scholar PubMed
Pan, H., Song, T., Wang, Z., Guo, Y., Zhang, H., Ji, T., Cao, K., and Zhang, Z. (2021). Ectopic BH3-only protein Bim acts as a cochaperone to positively regulate Hsp70 in yeast. J. Biochem. 170: 539–545, https://doi.org/10.1093/jb/mvab073.Suche in Google Scholar PubMed
Song, T., Guo, Y., Xue, Z., Guo, Z., Wang, Z., Lin, D., Zhang, H., Pan, H., Zhang, X., Yin, F., et al.. (2021). Small-molecule inhibitor targeting the Hsp70-Bim protein-protein interaction in CML cells overcomes BCR-ABL-independent TKI resistance. Leukemia 35: 2862–2874, https://doi.org/10.1038/s41375-021-01283-5.Suche in Google Scholar PubMed
Song, T., Yin, F., Wang, Z., Zhang, H., Liu, P., Guo, Y., Tang, Y., and Zhang, Z. (2023). Hsp70-Bim interaction facilitates mitophagy by recruiting parkin and TOMM20 into a complex. Cell. Mol. Biol. Lett. 28: 46, https://doi.org/10.1186/s11658-023-00458-5.Suche in Google Scholar PubMed PubMed Central
Tang, Q., Jin, M.W., Xiang, J.Z., Dong, M.Q., Sun, H.Y., Lau, C.P., and Li, G.R. (2007). The membrane permeable calcium chelator BAPTA-AM directly blocks human ether a-go-go-related gene potassium channels stably expressed in HEK 293 cells. Biochem. Pharmacol. 74: 1596–1607, https://doi.org/10.1016/j.bcp.2007.07.042.Suche in Google Scholar PubMed
Wallace, D.C., Brown, M.D., Melov, S., Graham, B., and Lott, M. (1998). Mitochondrial biology, degenerative diseases and aging. Biofactors 7: 187–190, https://doi.org/10.1002/biof.5520070303.Suche in Google Scholar PubMed
Wanderoy, S., Hees, J.T., Klesse, R., Edlich, F., and Harbauer, A.B. (2020). Kill one or kill the many: interplay between mitophagy and apoptosis. Biol. Chem. 402: 73–88, https://doi.org/10.1515/hsz-2020-0231.Suche in Google Scholar PubMed
Wang, S., Long, H., Hou, L., Feng, B., Ma, Z., Wu, Y., Zeng, Y., Cai, J., Zhang, D.W., and Zhao, G. (2023a). The mitophagy pathway and its implications in human diseases. Signal Transduct. Targeted Ther. 8: 304, https://doi.org/10.1038/s41392-023-01503-7.Suche in Google Scholar PubMed PubMed Central
Wang, Z., Song, T., Guo, Z., Uwituze, L.B., Guo, Y., Zhang, H., Wang, H., Zhang, X., Pan, H., Ji, T., et al.. (2021). A novel Hsp70 inhibitor specifically targeting the cancer-related Hsp70-Bim protein-protein interaction. Eur. J. Med. Chem. 220: 113452, https://doi.org/10.1016/j.ejmech.2021.113452.Suche in Google Scholar PubMed
Wang, Z., Zhang, H., Li, X., Song, Y., Wang, Y., Hu, Z., Gao, Q., Jiang, M., Yin, F., Yuan, L., et al.. (2023b). Exploiting the hot-spots of Hsp70-Bim protein-protein interaction to optimize the 1-oxo-1H-phenalene-2,3-dicarbonitrile analogues as specific Hsp70-Bim inhibitors. J. Med. Chem. 66: 16377–16387, https://doi.org/10.1021/acs.jmedchem.3c01783.Suche in Google Scholar PubMed
Yin, Z., Popelka, H., Lei, Y., Yang, Y., and Klionsky, D.J. (2020). The roles of ubiquitin in mediating autophagy. Cells 9: 2025, https://doi.org/10.3390/cells9092025.Suche in Google Scholar PubMed PubMed Central
Yuan, L., Wang, Y., Margulis, B.A., Song, T., Wang, Z., and Zhang, Z. (2023). Ectopic BH3-only protein Bim associates with Hsp70 to regulate yeast mitophagy. Dokl. Biochem. Biophys. 512: 292–299, https://doi.org/10.1134/s1607672923700485.Suche in Google Scholar
Zhang, Z., Song, T., Zhang, T., Gao, J., Wu, G., An, L., and Du, G. (2011). A novel BH3 mimetic S1 potently induces Bax/Bak-dependent apoptosis by targeting both Bcl-2 and Mcl-1. Int. J. Cancer 128: 1724–1735, https://doi.org/10.1002/ijc.25484.Suche in Google Scholar PubMed
© 2024 Walter de Gruyter GmbH, Berlin/Boston
Artikel in diesem Heft
- Frontmatter
- Research Articles/Short Communications
- Protein Structure and Function
- Structural homology of mite profilins to plant profilins is not indicative of allergic cross-reactivity
- Membranes, Lipids, Glycobiology
- Apolipoprotein E-containing lipoproteins and their extracellular interactions with LRP1 affect LPS-induced inflammation
- Cell Biology and Signaling
- Inhibition of Chk1 stimulates cytotoxic action of platinum-based drugs and TRAIL combination in human prostate cancer cells
- 18β-glycyrrhetinic acid alleviates radiation-induced skin injury by activating the Nrf2/HO-1 signaling pathway
- Yeast Bxi1/Ybh3 mediates conserved mitophagy and apoptosis in yeast and mammalian cells: convergence in Bcl-2 family
- Novel Techniques
- Integrated machine learning and multimodal data fusion for patho-phenotypic feature recognition in iPSC models of dilated cardiomyopathy
Artikel in diesem Heft
- Frontmatter
- Research Articles/Short Communications
- Protein Structure and Function
- Structural homology of mite profilins to plant profilins is not indicative of allergic cross-reactivity
- Membranes, Lipids, Glycobiology
- Apolipoprotein E-containing lipoproteins and their extracellular interactions with LRP1 affect LPS-induced inflammation
- Cell Biology and Signaling
- Inhibition of Chk1 stimulates cytotoxic action of platinum-based drugs and TRAIL combination in human prostate cancer cells
- 18β-glycyrrhetinic acid alleviates radiation-induced skin injury by activating the Nrf2/HO-1 signaling pathway
- Yeast Bxi1/Ybh3 mediates conserved mitophagy and apoptosis in yeast and mammalian cells: convergence in Bcl-2 family
- Novel Techniques
- Integrated machine learning and multimodal data fusion for patho-phenotypic feature recognition in iPSC models of dilated cardiomyopathy