CARD19 Interacts with Mitochondrial Contact Site and Cristae Organizing System Constituent Proteins and Regulates Cristae Morphology
CARD19 is a mitochondrial protein of unknown function. While CARD19 was originally reported to regulate TCR-dependent NF-κB activation via interaction with BCL10, this function is not recapitulated ex vivo in primary murine CD8<sup>+</sup> T cells. Here, we employ a combination of SIM, T...
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| Format: | Article |
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MDPI AG
2022-03-01
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| Online Access: | https://www.mdpi.com/2073-4409/11/7/1175 |
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| author | Kariana E. Rios Ming Zhou Nathaniel M. Lott Chelsi R. Beauregard Dennis P. McDaniel Thomas P. Conrads Brian C. Schaefer |
| author_facet | Kariana E. Rios Ming Zhou Nathaniel M. Lott Chelsi R. Beauregard Dennis P. McDaniel Thomas P. Conrads Brian C. Schaefer |
| author_sort | Kariana E. Rios |
| collection | DOAJ |
| description | CARD19 is a mitochondrial protein of unknown function. While CARD19 was originally reported to regulate TCR-dependent NF-κB activation via interaction with BCL10, this function is not recapitulated ex vivo in primary murine CD8<sup>+</sup> T cells. Here, we employ a combination of SIM, TEM, and confocal microscopy, along with proteinase K protection assays and proteomics approaches, to identify interacting partners of CARD19 in macrophages. Our data show that CARD19 is specifically localized to the outer mitochondrial membrane. Through deletion of functional domains, we demonstrate that both the distal C-terminus and transmembrane domain are required for mitochondrial targeting, whereas the CARD is not. Importantly, mass spectrometry analysis of 3×Myc-CARD19 immunoprecipitates reveals that CARD19 interacts with the components of the mitochondrial intermembrane bridge (MIB), consisting of mitochondrial contact site and cristae organizing system (MICOS) components MIC19, MIC25, and MIC60, and MICOS-interacting proteins SAMM50 and MTX2. These CARD19 interactions are in part dependent on a properly folded CARD. Consistent with previously reported phenotypes upon siRNA silencing of MICOS subunits, absence of CARD19 correlates with irregular cristae morphology. Based on these data, we propose that CARD19 is a previously unknown interacting partner of the MIB and the MIC19–MIC25–MIC60 MICOS subcomplex that regulates cristae morphology. |
| first_indexed | 2024-03-09T12:00:49Z |
| format | Article |
| id | doaj.art-8f981d76c573421587c0da605ad086c0 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T12:00:49Z |
| publishDate | 2022-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-8f981d76c573421587c0da605ad086c02023-11-30T23:04:30ZengMDPI AGCells2073-44092022-03-01117117510.3390/cells11071175CARD19 Interacts with Mitochondrial Contact Site and Cristae Organizing System Constituent Proteins and Regulates Cristae MorphologyKariana E. Rios0Ming Zhou1Nathaniel M. Lott2Chelsi R. Beauregard3Dennis P. McDaniel4Thomas P. Conrads5Brian C. Schaefer6Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USAWomen’s Health Integrated Research Center, Women’s Service Line, Inova Health System, Annandale, VA 22003, USADepartment of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USADepartment of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USABiomedical Instrumentation Center, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USAWomen’s Health Integrated Research Center, Women’s Service Line, Inova Health System, Annandale, VA 22003, USADepartment of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USACARD19 is a mitochondrial protein of unknown function. While CARD19 was originally reported to regulate TCR-dependent NF-κB activation via interaction with BCL10, this function is not recapitulated ex vivo in primary murine CD8<sup>+</sup> T cells. Here, we employ a combination of SIM, TEM, and confocal microscopy, along with proteinase K protection assays and proteomics approaches, to identify interacting partners of CARD19 in macrophages. Our data show that CARD19 is specifically localized to the outer mitochondrial membrane. Through deletion of functional domains, we demonstrate that both the distal C-terminus and transmembrane domain are required for mitochondrial targeting, whereas the CARD is not. Importantly, mass spectrometry analysis of 3×Myc-CARD19 immunoprecipitates reveals that CARD19 interacts with the components of the mitochondrial intermembrane bridge (MIB), consisting of mitochondrial contact site and cristae organizing system (MICOS) components MIC19, MIC25, and MIC60, and MICOS-interacting proteins SAMM50 and MTX2. These CARD19 interactions are in part dependent on a properly folded CARD. Consistent with previously reported phenotypes upon siRNA silencing of MICOS subunits, absence of CARD19 correlates with irregular cristae morphology. Based on these data, we propose that CARD19 is a previously unknown interacting partner of the MIB and the MIC19–MIC25–MIC60 MICOS subcomplex that regulates cristae morphology.https://www.mdpi.com/2073-4409/11/7/1175CARD19BinCARDMICOSCARD proteinscristaeMIB |
| spellingShingle | Kariana E. Rios Ming Zhou Nathaniel M. Lott Chelsi R. Beauregard Dennis P. McDaniel Thomas P. Conrads Brian C. Schaefer CARD19 Interacts with Mitochondrial Contact Site and Cristae Organizing System Constituent Proteins and Regulates Cristae Morphology Cells CARD19 BinCARD MICOS CARD proteins cristae MIB |
| title | CARD19 Interacts with Mitochondrial Contact Site and Cristae Organizing System Constituent Proteins and Regulates Cristae Morphology |
| title_full | CARD19 Interacts with Mitochondrial Contact Site and Cristae Organizing System Constituent Proteins and Regulates Cristae Morphology |
| title_fullStr | CARD19 Interacts with Mitochondrial Contact Site and Cristae Organizing System Constituent Proteins and Regulates Cristae Morphology |
| title_full_unstemmed | CARD19 Interacts with Mitochondrial Contact Site and Cristae Organizing System Constituent Proteins and Regulates Cristae Morphology |
| title_short | CARD19 Interacts with Mitochondrial Contact Site and Cristae Organizing System Constituent Proteins and Regulates Cristae Morphology |
| title_sort | card19 interacts with mitochondrial contact site and cristae organizing system constituent proteins and regulates cristae morphology |
| topic | CARD19 BinCARD MICOS CARD proteins cristae MIB |
| url | https://www.mdpi.com/2073-4409/11/7/1175 |
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