title: Dynamics of mitochondrial RNA-binding protein complex in Trypanosoma brucei and its petite mutant under optimized immobilization conditions.
abstract: There are a variety of complex metabolic processes ongoing simultaneously in the single, large mitochondrion of Trypanosoma brucei. Understanding the organellar environment and dynamics of mitochondrial proteins requires quantitative measurement in vivo. In this study, we have validated a method for immobilizing both procyclic stage (PS) and bloodstream stage (BS) T. brucei brucei with a high level of cell viability over several hours and verified its suitability for undertaking fluorescence recovery after photobleaching (FRAP), with mitochondrion-targeted yellow fluorescent protein (YFP). Next, we used this method for comparative analysis of the translational diffusion of mitochondrial RNA-binding protein 1 (MRP1) in the BS and in T. b. evansi. The latter flagellate is like petite mutant Saccharomyces cerevisiae because it lacks organelle-encoded nucleic acids. FRAP measurement of YFP-tagged MRP1 in both cell lines illuminated from a new perspective how the absence or presence of RNA affects proteins involved in mitochondrial RNA metabolism. This work represents the first attempt to examine this process in live trypanosomes.
authors: Institute of Parasitology, Biology Centre, Czech Academy of Sciences, České Budĕjovice (Budweis), Czech Republic Faculty of Sciences, University of South Bohemia, České Budĕjovice (Budweis), Czech Republic.
authors: Zhenqiu
authors: Z
authors: Huang
authors: Faculty of Sciences, University of South Bohemia, České Budĕjovice (Budweis), Czech Republic.
authors: Sabine
authors: S
authors: Kaltenbrunner
authors: Institute for Molecular Genetics, Czech Academy of Sciences, Prague, Czech Republic.
authors: Eva
authors: E
authors: Šimková
authors: Institute for Molecular Genetics, Czech Academy of Sciences, Prague, Czech Republic.
authors: David
authors: D
authors: Stanĕk
authors: Institute of Parasitology, Biology Centre, Czech Academy of Sciences, České Budĕjovice (Budweis), Czech Republic Faculty of Sciences, University of South Bohemia, České Budĕjovice (Budweis), Czech Republic jula@paru.cas.cz hassan@paru.cas.cz.
authors: Julius
authors: J
authors: Lukeš
authors: Institute of Parasitology, Biology Centre, Czech Academy of Sciences, České Budĕjovice (Budweis), Czech Republic Faculty of Sciences, University of South Bohemia, České Budĕjovice (Budweis), Czech Republic jula@paru.cas.cz hassan@paru.cas.cz.
authors: Hassan
authors: H
authors: Hashimi
mesh_terms: D002470:Cell Survival / Q000235:genetics; D024101:Mitochondrial Proteins / Q000235:genetics; D009154:Mutation; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D012313:RNA / Q000235:genetics*; D034622:RNA Interference; D000077278:RNA, Mitochondrial; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014346:Trypanosoma brucei brucei / Q000235:genetics*
chemical_list: D024101:Mitochondrial Proteins; D015800:Protozoan Proteins; D000077278:RNA, Mitochondrial; D016601:RNA-Binding Proteins; C104816:gBP21 protein, Trypanosoma brucei; D012313:RNA