The mitochondrial Cu+ transporter PiC2 (SLC25A3) is a target of MTF1 and contributes to the development of skeletal muscle in vitro
The loading of copper (Cu) into cytochrome c oxidase (COX) in mitochondria is essential for energy production in cells. Extensive studies have been performed to characterize mitochondrial cuproenzymes that contribute to the metallation of COX, such as Sco1, Sco2, and Cox17. However, limited informat...
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Frontiers Media S.A.
2022-11-01
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| Series: | Frontiers in Molecular Biosciences |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmolb.2022.1037941/full |
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| author | Cat McCann Michael Quinteros Ifeoluwa Adelugba Marcos N. Morgada Aida R. Castelblanco Emily J. Davis Antonio Lanzirotti Sarah J. Hainer Alejandro J. Vila Juan G. Navea Teresita Padilla-Benavides |
| author_facet | Cat McCann Michael Quinteros Ifeoluwa Adelugba Marcos N. Morgada Aida R. Castelblanco Emily J. Davis Antonio Lanzirotti Sarah J. Hainer Alejandro J. Vila Juan G. Navea Teresita Padilla-Benavides |
| author_sort | Cat McCann |
| collection | DOAJ |
| description | The loading of copper (Cu) into cytochrome c oxidase (COX) in mitochondria is essential for energy production in cells. Extensive studies have been performed to characterize mitochondrial cuproenzymes that contribute to the metallation of COX, such as Sco1, Sco2, and Cox17. However, limited information is available on the upstream mechanism of Cu transport and delivery to mitochondria, especially through Cu-impermeable membranes, in mammalian cells. The mitochondrial phosphate transporter SLC25A3, also known as PiC2, binds Cu+ and transports the ion through these membranes in eukaryotic cells, ultimately aiding in the metallation of COX. We used the well-established differentiation model of primary myoblasts derived from mouse satellite cells, wherein Cu availability is necessary for growth and maturation, and showed that PiC2 is a target of MTF1, and its expression is both induced during myogenesis and favored by Cu supplementation. PiC2 deletion using CRISPR/Cas9 showed that the transporter is required for proliferation and differentiation of primary myoblasts, as both processes are delayed upon PiC2 knock-out. The effects of PiC2 deletion were rescued by the addition of Cu to the growth medium, implying the deleterious effects of PiC2 knockout in myoblasts may be in part due to a failure to deliver sufficient Cu to the mitochondria, which can be compensated by other mitochondrial cuproproteins. Co-localization and co-immunoprecipitation of PiC2 and COX also suggest that PiC2 may participate upstream in the copper delivery chain into COX, as verified by in vitro Cu+-transfer experiments. These data indicate an important role for PiC2 in both the delivery of Cu to the mitochondria and COX, favoring the differentiation of primary myoblasts. |
| first_indexed | 2024-04-11T07:11:15Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2296-889X |
| language | English |
| last_indexed | 2024-04-11T07:11:15Z |
| publishDate | 2022-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Biosciences |
| spelling | doaj.art-68efc8fe18604b64aa27dc185ae288ff2022-12-22T04:38:12ZengFrontiers Media S.A.Frontiers in Molecular Biosciences2296-889X2022-11-01910.3389/fmolb.2022.10379411037941The mitochondrial Cu+ transporter PiC2 (SLC25A3) is a target of MTF1 and contributes to the development of skeletal muscle in vitroCat McCann0Michael Quinteros1Ifeoluwa Adelugba2Marcos N. Morgada3Aida R. Castelblanco4Emily J. Davis5Antonio Lanzirotti6Sarah J. Hainer7Alejandro J. Vila8Juan G. Navea9Teresita Padilla-Benavides10Department of Molecular Biology and Biochemistry, Wesleyan University, Middletown, CT, United StatesDepartment of Molecular Biology and Biochemistry, Wesleyan University, Middletown, CT, United StatesUniversity of Massachusetts Chan Medical School, Worcester, MA, United StatesInstituto de Biología Molecular y Celular de Rosario, Rosario, ArgentinaDepartment of Chemistry, Skidmore College, Saratoga Springs, NY, United StatesDepartment of Chemistry, Skidmore College, Saratoga Springs, NY, United StatesCenter for Advanced Radiation Sources, The University of Chicago, Chicago, IL, United StatesDepartment of Biological Sciences. University of Pittsburgh, Pittsburgh, PA, United StatesInstituto de Biología Molecular y Celular de Rosario, Rosario, ArgentinaDepartment of Chemistry, Skidmore College, Saratoga Springs, NY, United StatesDepartment of Molecular Biology and Biochemistry, Wesleyan University, Middletown, CT, United StatesThe loading of copper (Cu) into cytochrome c oxidase (COX) in mitochondria is essential for energy production in cells. Extensive studies have been performed to characterize mitochondrial cuproenzymes that contribute to the metallation of COX, such as Sco1, Sco2, and Cox17. However, limited information is available on the upstream mechanism of Cu transport and delivery to mitochondria, especially through Cu-impermeable membranes, in mammalian cells. The mitochondrial phosphate transporter SLC25A3, also known as PiC2, binds Cu+ and transports the ion through these membranes in eukaryotic cells, ultimately aiding in the metallation of COX. We used the well-established differentiation model of primary myoblasts derived from mouse satellite cells, wherein Cu availability is necessary for growth and maturation, and showed that PiC2 is a target of MTF1, and its expression is both induced during myogenesis and favored by Cu supplementation. PiC2 deletion using CRISPR/Cas9 showed that the transporter is required for proliferation and differentiation of primary myoblasts, as both processes are delayed upon PiC2 knock-out. The effects of PiC2 deletion were rescued by the addition of Cu to the growth medium, implying the deleterious effects of PiC2 knockout in myoblasts may be in part due to a failure to deliver sufficient Cu to the mitochondria, which can be compensated by other mitochondrial cuproproteins. Co-localization and co-immunoprecipitation of PiC2 and COX also suggest that PiC2 may participate upstream in the copper delivery chain into COX, as verified by in vitro Cu+-transfer experiments. These data indicate an important role for PiC2 in both the delivery of Cu to the mitochondria and COX, favoring the differentiation of primary myoblasts.https://www.frontiersin.org/articles/10.3389/fmolb.2022.1037941/fullSLC25A3PiC2MTF1copper transportcytochrome c oxidasemitochondria |
| spellingShingle | Cat McCann Michael Quinteros Ifeoluwa Adelugba Marcos N. Morgada Aida R. Castelblanco Emily J. Davis Antonio Lanzirotti Sarah J. Hainer Alejandro J. Vila Juan G. Navea Teresita Padilla-Benavides The mitochondrial Cu+ transporter PiC2 (SLC25A3) is a target of MTF1 and contributes to the development of skeletal muscle in vitro Frontiers in Molecular Biosciences SLC25A3 PiC2 MTF1 copper transport cytochrome c oxidase mitochondria |
| title | The mitochondrial Cu+ transporter PiC2 (SLC25A3) is a target of MTF1 and contributes to the development of skeletal muscle in vitro |
| title_full | The mitochondrial Cu+ transporter PiC2 (SLC25A3) is a target of MTF1 and contributes to the development of skeletal muscle in vitro |
| title_fullStr | The mitochondrial Cu+ transporter PiC2 (SLC25A3) is a target of MTF1 and contributes to the development of skeletal muscle in vitro |
| title_full_unstemmed | The mitochondrial Cu+ transporter PiC2 (SLC25A3) is a target of MTF1 and contributes to the development of skeletal muscle in vitro |
| title_short | The mitochondrial Cu+ transporter PiC2 (SLC25A3) is a target of MTF1 and contributes to the development of skeletal muscle in vitro |
| title_sort | mitochondrial cu transporter pic2 slc25a3 is a target of mtf1 and contributes to the development of skeletal muscle in vitro |
| topic | SLC25A3 PiC2 MTF1 copper transport cytochrome c oxidase mitochondria |
| url | https://www.frontiersin.org/articles/10.3389/fmolb.2022.1037941/full |
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