Abstract
MitoNEET is an outer mitochondrial membrane protein essential for sensing and regulation of iron and reactive oxygen species (ROS) homeostasis. It is a key player in multiple human maladies including diabetes, cancer, neurodegeneration, and Parkinson’s diseases. In healthy cells, mitoNEET receives its clusters from the mitochondrion and transfers them to acceptor proteins in a process that could be altered by drugs or during illness. Here, we report that mitoNEET regulates the outer-mitochondrial membrane (OMM) protein voltage-dependent anion channel 1 (VDAC1). VDAC1 is a crucial player in the cross talk between the mitochondria and the cytosol. VDAC proteins function to regulate metabolites, ions, ROS, and fatty acid transport, as well as function as a “governator” sentry for the transport of metabolites and ions between the cytosol and the mitochondria. We find that the redox-sensitive [2Fe-2S] cluster protein mitoNEET gates VDAC1 when mitoNEET is oxidized. Addition of the VDAC inhibitor 4,4′-diisothiocyanatostilbene-2,2′-disulfonate (DIDS) prevents both mitoNEET binding in vitro and mitoNEET-dependent mitochondrial iron accumulation in situ. We find that the DIDS inhibitor does not alter the redox state of MitoNEET. Taken together, our data indicate that mitoNEET regulates VDAC in a redox-dependent manner in cells, closing the pore and likely disrupting VDAC’s flow of metabolites.
| Original language | American English |
|---|---|
| Pages (from-to) | 19924-19929 |
| Number of pages | 6 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 116 |
| Issue number | 40 |
| DOIs | |
| State | Published - 1 Oct 2019 |
Bibliographical note
Funding Information:ACKNOWLEDGMENTS. This work was supported by NSF–Binational Science Foundation (BSF) funding (NSF-MCB-1613462 [R.M.]) and BSF (BSF Grant 2015831 [R.N.]). Work at the laboratory of P.A.J. is supported by National Institutes of Health Grant GM101467. J.N.O. is a Cancer Prevention and Research Institute of Texas (CPRIT) Scholar in Cancer Research sponsored by CPRIT. J.N.O. was supported by the Center for Theoretical Biological Physics sponsored by the NSF (Grant PHY-1427654) and by NSF-CHE 1614101. The funders had no role in the design, data collection, analysis, decision to publish, or preparation of the manuscript. We also thank Prof. Varda Shoshan-Barmatz from Ben Gurion University, who generously shared her VDAC conducting system to collect data that was used in Fig. 2, as well as Prof. Emmanuel Theodorakis and Dr. Kendra Hailey from University of California at San Diego for their thoughtful as well as valuable insights into this manuscript.
Funding Information:
This work was supported by NSF?Binational Science Foundation (BSF) funding (NSF-MCB-1613462 [R.M.]) and BSF (BSF Grant 2015831 [R.N.]). Work at the laboratory of P.A.J. is supported by National Institutes of Health Grant GM101467. J.N.O. is a Cancer Prevention and Research Institute of Texas (CPRIT) Scholar in Cancer Research sponsored by CPRIT. J.N.O. was supported by the Center for Theoretical Biological Physics sponsored by the NSF (Grant PHY-1427654) and by NSF-CHE 1614101. The funders had no role in the design, data collection, analysis, decision to publish, or preparation of the manuscript. We also thank Prof. Varda Shoshan-Barmatz from Ben Gurion University, who generously shared her VDAC conducting system to collect data that was used in Fig. 2, as well as Prof. Emmanuel Theodorakis and Dr. Kendra Hailey from University of California at San Diego for their thoughtful as well as valuable insights into this manuscript.
Publisher Copyright:
© 2019 National Academy of Sciences. All rights reserved.
Keywords
- CISD1
- Direct coupling
- Ferroptosis
- MitoNEET
- VDAC1
