Mitochondrial metabolic remodeling is a hallmark of the Trypanosoma brucei digenetic life cycle because the insect stage utilizes a cost-effective oxidative phosphorylation (OxPhos) to generate ATP, while bloodstream cells switch to aerobic glycolysis. Due to difficulties in acquiring enough parasites from the tsetse fly vector, the dynamics of the parasite's metabolic rewiring in the vector have remained obscure. Here, we took advantage of in vitro-induced differentiation to follow changes at the RNA, protein, and metabolite levels. This multi-omics and cell-based profiling showed an immediate redirection of electron flow from the cytochrome-mediated pathway to an alternative oxidase (AOX), an increase in proline consumption, elevated activity of complex II, and certain tricarboxylic acid (TCA) cycle enzymes, which led to mitochondrial membrane hyperpolarization and increased reactive oxygen species (ROS) levels. Interestingly, these ROS molecules appear to act as signaling molecules driving developmental progression because ectopic expression of catalase, a ROS scavenger, halted the in vitro-induced differentiation. Our results provide insights into the mechanisms of the parasite's mitochondrial rewiring and reinforce the emerging concept that mitochondria act as signaling organelles through release of ROS to drive cellular differentiation.
- MeSH
- adenosintrifosfát biosyntéza MeSH
- buněčná diferenciace účinky léků MeSH
- buněčné dýchání účinky léků MeSH
- buněčné linie MeSH
- elektrony MeSH
- glukosa farmakologie MeSH
- membránový potenciál mitochondrií účinky léků MeSH
- metabolické sítě a dráhy účinky léků MeSH
- metabolomika * MeSH
- mitochondriální proteiny metabolismus MeSH
- mitochondrie účinky léků metabolismus MeSH
- oxidace-redukce MeSH
- oxidoreduktasy metabolismus MeSH
- prolin metabolismus MeSH
- proteom metabolismus MeSH
- protozoální proteiny metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- rostlinné proteiny metabolismus MeSH
- signální transdukce MeSH
- transkriptom genetika MeSH
- transport elektronů účinky léků MeSH
- Trypanosoma brucei brucei účinky léků genetika růst a vývoj metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Trypanosomes are masters of adaptation to different host environments during their complex life cycle. Large-scale proteomic approaches provide information on changes at the cellular level, and in a systematic way. However, detailed work on single components is necessary to understand the adaptation mechanisms on a molecular level. Here, we have performed a detailed characterization of a bloodstream form (BSF) stage-specific putative flagellar host adaptation factor Tb927.11.2400, identified previously in a SILAC-based comparative proteome study. Tb927.11.2400 shares 38% amino acid identity with TbFlabarin (Tb927.11.2410), a procyclic form (PCF) stage-specific flagellar BAR domain protein. We named Tb927.11.2400 TbFlabarin-like (TbFlabarinL), and demonstrate that it originates from a gene duplication event, which occurred in the African trypanosomes. TbFlabarinL is not essential for the growth of the parasites under cell culture conditions and it is dispensable for developmental differentiation from BSF to the PCF in vitro. We generated TbFlabarinL-specific antibodies, and showed that it localizes in the flagellum. Co-immunoprecipitation experiments together with a biochemical cell fractionation suggest a dual association of TbFlabarinL with the flagellar membrane and the components of the paraflagellar rod.
- MeSH
- duplikace genu MeSH
- flagella fyziologie MeSH
- fylogeneze MeSH
- geneticky modifikované organismy MeSH
- myši inbrední C57BL MeSH
- proteinové domény MeSH
- protozoální proteiny chemie genetika imunologie metabolismus MeSH
- Trypanosoma brucei brucei genetika patogenita fyziologie MeSH
- trypanozomóza africká parazitologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
