Unique Interactions of the Small Translocases of the Mitochondrial Inner Membrane (Tims) in <i>Trypanosoma brucei</i>
The infectious agent for African trypanosomiasis, <i>Trypanosoma brucei</i>, possesses a unique and essential translocase of the mitochondrial inner membrane, known as the TbTIM17 complex. TbTim17 associates with six small TbTims (TbTim9, TbTim10, TbTim11, TbTim12, TbTim13, and TbTim8/13...
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MDPI AG
2024-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/25/3/1415 |
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| author | Linda S. Quiñones Fidel Soto Gonzalez Chauncey Darden Muhammad Khan Anuj Tripathi Joseph T. Smith Jamaine Davis Smita Misra Minu Chaudhuri |
| author_facet | Linda S. Quiñones Fidel Soto Gonzalez Chauncey Darden Muhammad Khan Anuj Tripathi Joseph T. Smith Jamaine Davis Smita Misra Minu Chaudhuri |
| author_sort | Linda S. Quiñones |
| collection | DOAJ |
| description | The infectious agent for African trypanosomiasis, <i>Trypanosoma brucei</i>, possesses a unique and essential translocase of the mitochondrial inner membrane, known as the TbTIM17 complex. TbTim17 associates with six small TbTims (TbTim9, TbTim10, TbTim11, TbTim12, TbTim13, and TbTim8/13). However, the interaction patterns of these smaller TbTims with each other and TbTim17 are not clear. Through yeast two-hybrid (Y2H) and co-immunoprecipitation analyses, we demonstrate that all six small TbTims interact with each other. Stronger interactions were found among TbTim8/13, TbTim9, and TbTim10. However, TbTim10 shows weaker associations with TbTim13, which has a stronger connection with TbTim17. Each of the small TbTims also interacts strongly with the C-terminal region of TbTim17. RNAi studies indicated that among all small TbTims, TbTim13 is most crucial for maintaining the steady-state levels of the TbTIM17 complex. Further analysis of the small TbTim complexes by size exclusion chromatography revealed that each small TbTim, except for TbTim13, is present in ~70 kDa complexes, possibly existing in heterohexameric forms. In contrast, TbTim13 is primarily present in the larger complex (>800 kDa) and co-fractionates with TbTim17. Altogether, our results demonstrate that, relative to other eukaryotes, the architecture and function of the small TbTim complexes are specific to <i>T. brucei</i>. |
| first_indexed | 2024-03-08T03:56:57Z |
| format | Article |
| id | doaj.art-b37a1bd789c2452895cc37d585df10b8 |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-08T03:56:57Z |
| publishDate | 2024-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-b37a1bd789c2452895cc37d585df10b82024-02-09T15:13:09ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-01-01253141510.3390/ijms25031415Unique Interactions of the Small Translocases of the Mitochondrial Inner Membrane (Tims) in <i>Trypanosoma brucei</i>Linda S. Quiñones0Fidel Soto Gonzalez1Chauncey Darden2Muhammad Khan3Anuj Tripathi4Joseph T. Smith5Jamaine Davis6Smita Misra7Minu Chaudhuri8Department of Microbiology, Immunology, and Physiology, School of Medicine, Meharry Medical College, Nashville, TN 37208, USADepartment of Microbiology, Immunology, and Physiology, School of Medicine, Meharry Medical College, Nashville, TN 37208, USADepartment of Biochemistry, Cancer Biology, Neuroscience, and Pharmacology, School of Medicine, Meharry Medical College, Nashville, TN 37208, USADepartment of Microbiology, Immunology, and Physiology, School of Medicine, Meharry Medical College, Nashville, TN 37208, USADepartment of Microbiology, Immunology, and Physiology, School of Medicine, Meharry Medical College, Nashville, TN 37208, USADepartment of Microbiology and Immunology, Jacobs School of Medicine and Biomedical Sciences, University at Buffalo, Buffalo, NY 14203, USADepartment of Biochemistry, Cancer Biology, Neuroscience, and Pharmacology, School of Medicine, Meharry Medical College, Nashville, TN 37208, USADepartment of Biomedical Science, School of Graduate Studies, Meharry Medical College, Nashville, TN 37208, USADepartment of Microbiology, Immunology, and Physiology, School of Medicine, Meharry Medical College, Nashville, TN 37208, USAThe infectious agent for African trypanosomiasis, <i>Trypanosoma brucei</i>, possesses a unique and essential translocase of the mitochondrial inner membrane, known as the TbTIM17 complex. TbTim17 associates with six small TbTims (TbTim9, TbTim10, TbTim11, TbTim12, TbTim13, and TbTim8/13). However, the interaction patterns of these smaller TbTims with each other and TbTim17 are not clear. Through yeast two-hybrid (Y2H) and co-immunoprecipitation analyses, we demonstrate that all six small TbTims interact with each other. Stronger interactions were found among TbTim8/13, TbTim9, and TbTim10. However, TbTim10 shows weaker associations with TbTim13, which has a stronger connection with TbTim17. Each of the small TbTims also interacts strongly with the C-terminal region of TbTim17. RNAi studies indicated that among all small TbTims, TbTim13 is most crucial for maintaining the steady-state levels of the TbTIM17 complex. Further analysis of the small TbTim complexes by size exclusion chromatography revealed that each small TbTim, except for TbTim13, is present in ~70 kDa complexes, possibly existing in heterohexameric forms. In contrast, TbTim13 is primarily present in the larger complex (>800 kDa) and co-fractionates with TbTim17. Altogether, our results demonstrate that, relative to other eukaryotes, the architecture and function of the small TbTim complexes are specific to <i>T. brucei</i>.https://www.mdpi.com/1422-0067/25/3/1415mitochondriatranslocasesmall Timsprotein interactions<i>Trypanosoma brucei</i> |
| spellingShingle | Linda S. Quiñones Fidel Soto Gonzalez Chauncey Darden Muhammad Khan Anuj Tripathi Joseph T. Smith Jamaine Davis Smita Misra Minu Chaudhuri Unique Interactions of the Small Translocases of the Mitochondrial Inner Membrane (Tims) in <i>Trypanosoma brucei</i> International Journal of Molecular Sciences mitochondria translocase small Tims protein interactions <i>Trypanosoma brucei</i> |
| title | Unique Interactions of the Small Translocases of the Mitochondrial Inner Membrane (Tims) in <i>Trypanosoma brucei</i> |
| title_full | Unique Interactions of the Small Translocases of the Mitochondrial Inner Membrane (Tims) in <i>Trypanosoma brucei</i> |
| title_fullStr | Unique Interactions of the Small Translocases of the Mitochondrial Inner Membrane (Tims) in <i>Trypanosoma brucei</i> |
| title_full_unstemmed | Unique Interactions of the Small Translocases of the Mitochondrial Inner Membrane (Tims) in <i>Trypanosoma brucei</i> |
| title_short | Unique Interactions of the Small Translocases of the Mitochondrial Inner Membrane (Tims) in <i>Trypanosoma brucei</i> |
| title_sort | unique interactions of the small translocases of the mitochondrial inner membrane tims in i trypanosoma brucei i |
| topic | mitochondria translocase small Tims protein interactions <i>Trypanosoma brucei</i> |
| url | https://www.mdpi.com/1422-0067/25/3/1415 |
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