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Characterization of two novel proteins involved in mitochondrial DNA anchoring in Trypanosoma brucei
Trypanosoma brucei is a single celled eukaryotic parasite in the group of the Kinetoplastea. The parasite harbors a single mitochondrion with a singular mitochondrial genome that is known as the kinetoplast DNA (kDNA). The kDNA consists of a unique network of thousands of interlocked circular DNA mo...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10374059/ https://www.ncbi.nlm.nih.gov/pubmed/37459364 http://dx.doi.org/10.1371/journal.ppat.1011486 |
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author | Amodeo, Simona Bregy, Irina Hoffmann, Anneliese Fradera-Sola, Albert Kern, Mara Baudouin, Hélène Zuber, Benoît Butter, Falk Ochsenreiter, Torsten |
author_facet | Amodeo, Simona Bregy, Irina Hoffmann, Anneliese Fradera-Sola, Albert Kern, Mara Baudouin, Hélène Zuber, Benoît Butter, Falk Ochsenreiter, Torsten |
author_sort | Amodeo, Simona |
collection | PubMed |
description | Trypanosoma brucei is a single celled eukaryotic parasite in the group of the Kinetoplastea. The parasite harbors a single mitochondrion with a singular mitochondrial genome that is known as the kinetoplast DNA (kDNA). The kDNA consists of a unique network of thousands of interlocked circular DNA molecules. To ensure proper inheritance of the kDNA to the daughter cells, the genome is physically linked to the basal body, the master organizer of the cell cycle in trypanosomes. The connection that spans, cytoplasm, mitochondrial membranes and the mitochondrial matrix is mediated by the Tripartite Attachment Complex (TAC). Using a combination of proteomics and RNAi we test the current model of hierarchical TAC assembly and identify TbmtHMG44 and TbKAP68 as novel candidates of a complex that connects the TAC to the kDNA. Depletion of TbmtHMG44 or TbKAP68 each leads to a strong kDNA loss but not missegregation phenotype as previously defined for TAC components. We demonstrate that the proteins rely on both the TAC and the kDNA for stable localization to the interface between these two structures. In vitro experiments suggest a direct interaction between TbmtHMG44 and TbKAP68 and that recombinant TbKAP68 is a DNA binding protein. We thus propose that TbmtHMG44 and TbKAP68 are part of a distinct complex connecting the kDNA to the TAC. |
format | Online Article Text |
id | pubmed-10374059 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-103740592023-07-28 Characterization of two novel proteins involved in mitochondrial DNA anchoring in Trypanosoma brucei Amodeo, Simona Bregy, Irina Hoffmann, Anneliese Fradera-Sola, Albert Kern, Mara Baudouin, Hélène Zuber, Benoît Butter, Falk Ochsenreiter, Torsten PLoS Pathog Research Article Trypanosoma brucei is a single celled eukaryotic parasite in the group of the Kinetoplastea. The parasite harbors a single mitochondrion with a singular mitochondrial genome that is known as the kinetoplast DNA (kDNA). The kDNA consists of a unique network of thousands of interlocked circular DNA molecules. To ensure proper inheritance of the kDNA to the daughter cells, the genome is physically linked to the basal body, the master organizer of the cell cycle in trypanosomes. The connection that spans, cytoplasm, mitochondrial membranes and the mitochondrial matrix is mediated by the Tripartite Attachment Complex (TAC). Using a combination of proteomics and RNAi we test the current model of hierarchical TAC assembly and identify TbmtHMG44 and TbKAP68 as novel candidates of a complex that connects the TAC to the kDNA. Depletion of TbmtHMG44 or TbKAP68 each leads to a strong kDNA loss but not missegregation phenotype as previously defined for TAC components. We demonstrate that the proteins rely on both the TAC and the kDNA for stable localization to the interface between these two structures. In vitro experiments suggest a direct interaction between TbmtHMG44 and TbKAP68 and that recombinant TbKAP68 is a DNA binding protein. We thus propose that TbmtHMG44 and TbKAP68 are part of a distinct complex connecting the kDNA to the TAC. Public Library of Science 2023-07-17 /pmc/articles/PMC10374059/ /pubmed/37459364 http://dx.doi.org/10.1371/journal.ppat.1011486 Text en © 2023 Amodeo et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Amodeo, Simona Bregy, Irina Hoffmann, Anneliese Fradera-Sola, Albert Kern, Mara Baudouin, Hélène Zuber, Benoît Butter, Falk Ochsenreiter, Torsten Characterization of two novel proteins involved in mitochondrial DNA anchoring in Trypanosoma brucei |
title | Characterization of two novel proteins involved in mitochondrial DNA anchoring in Trypanosoma brucei |
title_full | Characterization of two novel proteins involved in mitochondrial DNA anchoring in Trypanosoma brucei |
title_fullStr | Characterization of two novel proteins involved in mitochondrial DNA anchoring in Trypanosoma brucei |
title_full_unstemmed | Characterization of two novel proteins involved in mitochondrial DNA anchoring in Trypanosoma brucei |
title_short | Characterization of two novel proteins involved in mitochondrial DNA anchoring in Trypanosoma brucei |
title_sort | characterization of two novel proteins involved in mitochondrial dna anchoring in trypanosoma brucei |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10374059/ https://www.ncbi.nlm.nih.gov/pubmed/37459364 http://dx.doi.org/10.1371/journal.ppat.1011486 |
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