This study analyses and characterises novel mitochondrial proteins in the parasitic protist Trypanosoma brucei. Applying phylogenetic analysis was described the evolutionary origin of ZapE protein in eukaryotes, using a newly developed proximity-dependent biotinylation approach (BioID2) we identified ZapE interaction partners like Oxa1. We also discovered a relationship when distribution of mitochondrial ZapE is restricted only to organisms with Oxa1, respiratory complexes, and a mitochondrial genome. TbPams were detected by phylogenetic analyses as orthologs of corresponding proteins in Opistokonts. We analyse the function of TbPam18 and TbPam16 in the replication of the mitochondrial DNA and determine, how the TMDs of TbPam18 and TbPam16 are essential for their functions. Finally, we evaluated a set of putative mitochondrial proteins of the heterolobosean N. gruberi defined by Localisation of Organelle Proteins by Isotope Tagging (LOPIT) and analyse the origin of mtFfh and mtFtsY.
Anotace v angličtině
This study analyses and characterises novel mitochondrial proteins in the parasitic protist Trypanosoma brucei. Applying phylogenetic analysis was described the evolutionary origin of ZapE protein in eukaryotes, using a newly developed proximity-dependent biotinylation approach (BioID2) we identified ZapE interaction partners like Oxa1. We also discovered a relationship when distribution of mitochondrial ZapE is restricted only to organisms with Oxa1, respiratory complexes, and a mitochondrial genome. TbPams were detected by phylogenetic analyses as orthologs of corresponding proteins in Opistokonts. We analyse the function of TbPam18 and TbPam16 in the replication of the mitochondrial DNA and determine, how the TMDs of TbPam18 and TbPam16 are essential for their functions. Finally, we evaluated a set of putative mitochondrial proteins of the heterolobosean N. gruberi defined by Localisation of Organelle Proteins by Isotope Tagging (LOPIT) and analyse the origin of mtFfh and mtFtsY.
This study analyses and characterises novel mitochondrial proteins in the parasitic protist Trypanosoma brucei. Applying phylogenetic analysis was described the evolutionary origin of ZapE protein in eukaryotes, using a newly developed proximity-dependent biotinylation approach (BioID2) we identified ZapE interaction partners like Oxa1. We also discovered a relationship when distribution of mitochondrial ZapE is restricted only to organisms with Oxa1, respiratory complexes, and a mitochondrial genome. TbPams were detected by phylogenetic analyses as orthologs of corresponding proteins in Opistokonts. We analyse the function of TbPam18 and TbPam16 in the replication of the mitochondrial DNA and determine, how the TMDs of TbPam18 and TbPam16 are essential for their functions. Finally, we evaluated a set of putative mitochondrial proteins of the heterolobosean N. gruberi defined by Localisation of Organelle Proteins by Isotope Tagging (LOPIT) and analyse the origin of mtFfh and mtFtsY.
Anotace v angličtině
This study analyses and characterises novel mitochondrial proteins in the parasitic protist Trypanosoma brucei. Applying phylogenetic analysis was described the evolutionary origin of ZapE protein in eukaryotes, using a newly developed proximity-dependent biotinylation approach (BioID2) we identified ZapE interaction partners like Oxa1. We also discovered a relationship when distribution of mitochondrial ZapE is restricted only to organisms with Oxa1, respiratory complexes, and a mitochondrial genome. TbPams were detected by phylogenetic analyses as orthologs of corresponding proteins in Opistokonts. We analyse the function of TbPam18 and TbPam16 in the replication of the mitochondrial DNA and determine, how the TMDs of TbPam18 and TbPam16 are essential for their functions. Finally, we evaluated a set of putative mitochondrial proteins of the heterolobosean N. gruberi defined by Localisation of Organelle Proteins by Isotope Tagging (LOPIT) and analyse the origin of mtFfh and mtFtsY.
The defense was held in hybrid format, in person and online via Zoom - committee: prof. RNDr. Ivo Šauman, CSc. – head, in person,RNDr. Ladislav Anděra, CSc. - ONLINE, prof. RNDr. Jan Tachezy, CSc.- ONLINE, doc. RNDr. Alena Panicucci Zíková, Ph.D. - in person, reviewers: Dr. Dave Speijer, Ph.D. (University of Amsterdam) - ONLINE, prof. MSc. Vjačeslav Jurčenko, Ph.D. (Ostravská univerzita) - in person. 25 people attended the defense in person.
Defense:
1. Co-supervisor Juliu Lukeš (originally supervisor) read his statement.
2. Supervisor Ignatio Durante read his statement.
3. Student presented her work in approx. 30 min.
4. Reviewer Dave Speijer commented his review and presentation and discuss with the student: 1. How could the studied proteins obtain completely new function? (Dave added that it is very difficult general question) - student hesitated and answered rather simply. 2. Figure 3 (page 114) - knock down effect on proteomics - side effect of low ATP on maxicircles as alternative explanation? Student repeated their explanation and defended that. Dave was mostly satisfied with answers.
5. Reviewer Vjačeslav Jurčenko read his review and discussed with the student his specific questions from his report. Student answered all of them.
6. Discussion with committe - A. Zikova asked couple questions, all answered. J. Tachezy - commneted Dave's question and added question to transportation mechanisms in mitochondria - student again rather hesitated and admitted that she did not think about that. (L. Andera had to leave slightly before the end of the discussion and voted online).
7. Without public - discussion about defense and voting.
VOTES - 3 in person and 3 ONLINE via MS FORMS: Valid votes - 6, Passed - 5 Failed - 1