Molecular model of the mitochondrial genome segregation machinery in
            <i>Trypanosoma brucei</i>

Hoffmann, Anneliese; Käser, Sandro; Jakob, Martin; Amodeo, Simona; Peitsch, Camille Françoise; Týč, Jiří; Vaughan, Sue; Zuber, Benoît; Schneider, André; Ochsenreiter, Torsten

doi:10.1073/pnas.1716582115

Cited by 43 publications

(105 citation statements)

References 61 publications

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“…A total of 138 million interactions were screened and 15 clones were isolated and characterised (Table S3). Three high confidence hits were identified expressing a N-terminal part of the TAC component p166 ( (Zhao et al 2008;Hoffmann et al 2018) ). This large, acidic TAC protein contains a single transmembrane domain in the C-terminus and was previously shown by superresolution microscopy to be in close proximity to TAC102 (Zhao et al 2008;Hoffmann et al 2018) .…”

Section: Yeast Two-hybrid Screen Shows Interaction Between Tac102 Andmentioning

confidence: 99%

“…Three high confidence hits were identified expressing a N-terminal part of the TAC component p166 ( (Zhao et al 2008;Hoffmann et al 2018) ). This large, acidic TAC protein contains a single transmembrane domain in the C-terminus and was previously shown by superresolution microscopy to be in close proximity to TAC102 (Zhao et al 2008;Hoffmann et al 2018) . Two of the three high confidence p166 clones expressed a short region from amino acid 37 to 210 while the third clone expressed a larger fragment ranging from amino acid 71 to 716 of the 1502 amino acid long protein ( Figure S4).…”

Section: Yeast Two-hybrid Screen Shows Interaction Between Tac102 Andmentioning

confidence: 99%

“…A number of additional components that display multiple localisations including in the TAC, like the E2 subunit of the α-ketoglutarate dehydrogenase and the tubulin-binding cofactor C protein, have been identified (Sykes and Hajduk 2013;André et al 2013) . The assembly of the TAC occurs de novo from the base of the flagellum towards the kDNA in a hierarchical manner such that kDNA proximal components depend on the proper assembly of the kDNA distal components (Hoffmann et al 2018;Schneider and Ochsenreiter 2018) . In order to identify novel interactors of TAC102, we used three different approaches with TAC102 as the bait: (i) proximity-dependent biotin identification (BioID), (ii) immunoprecipitation with a monoclonal TAC102 antibody and (iii) yeast two-hybrid.…”

Section: Introductionmentioning

confidence: 99%

“…In this study we used a monoclonal antibody raised against a region in the C-terminus of TAC102. This antibody was previously shown to be highly specific for TAC102 in fixed cells and native/denatured protein extracts (Trikin et al 2016;Hoffmann et al 2018) . The third approach we applied was a yeast two-hybrid screens that relies on the reconstitution of a functional transcription factor when two peptides of interest interact (Fields and Song 1989) .…”

Section: Introductionmentioning

confidence: 99%

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Discovery of new mitochondrial DNA segregation machinery components inTrypanosoma brucei: a comparison of three approaches

Hcm

Pfeiffer

Ochsenreiter

2019

Preprint

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Trypanosoma brucei is a single celled eukaryotic parasite and the causative agent of human African sleeping sickness and Nagana in cattle. Aside from its medical relevance T. brucei has also been key to the discovery of several general biological principles including GPI-anchoring, RNA-editing and trans-splicing. The parasite contains a single mitochondrial organelle with a singular genome. Recent studies have identified several molecular components of the mitochondrial genome segregation machinery (tripartite attachment complex, TAC), which connects the basal body of the flagellum to the mitochondrial DNA of T. brucei . The TAC component in closest proximity to the mitochondrial DNA is TAC102.Here we apply and compare three different approaches (proximity labeling, Immunoprecipitation and yeast two-hybrid) to identify novel interactors of TAC102 and subsequently verify their localisation. Furthermore, we establish the direct interaction of TAC102 and p166 in the unilateral filaments of the TAC.

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Section: Yeast Two-hybrid Screen Shows Interaction Between Tac102 Andmentioning

confidence: 99%

Section: Yeast Two-hybrid Screen Shows Interaction Between Tac102 Andmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Discovery of new mitochondrial DNA segregation machinery components inTrypanosoma brucei: a comparison of three approaches

Hcm

Pfeiffer

Ochsenreiter

2019

Preprint

Self Cite

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“…have not yet been investigated (Hoffmann et al, 2018;Trikin et al, 2016; Zhao, Lindsay, Roy Chowdhury, Robinson, & Englund, 2008).…”

Section: Cells With Two Temporally Distinct Flagella: Age Discriminmentioning

confidence: 99%

Dealing with several flagella in the same cell

Bertiaux

Bastin

2020

Cellular Microbiology

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Flagella are sophisticated organelles found in many eukaryotic microbes where they perform functions related to motility, signal detection, or cell morphogenesis. In many cases, several flagella are present per cell, and these can have a different composition, length, age, or function, raising the question of how this is managed. When the flagella are equivalent and constructed simultaneously such as in Chlamydomonas or Naegleria, we propose an equal access model where molecular components have free access to each organelle. By contrast, Trypanosoma and Leishmania contain temporally distinct organelles and elongate a new flagellum whilst maintaining the existing one. The equal access model could function providing that the mature flagellum is "locked" so that it can no longer be elongated or shortened. Alternatively, access of flagellar components could be restricted at the level of the basal body, the transition zone, or the loading on intraflagellar transport trains. In organisms that contains flagella of different age and composition such as Giardia, a temporal dimension is necessary, with the production of protein components of flagella spreading over one or more cell cycles. In the future, deciphering the molecular mechanisms involved in these processes should reveal new insights in flagellum assembly and function.

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Airyscan Superresolution Microscopy to Study Trypanosomatid Cell Biology

Harmer

Belbelazi

Carr

et al. 2020

Methods in Molecular Biology

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For over 35 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-bystep fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice. These hallmark features were introduced by series editor Dr. John Walker and constitute the key ingredient in each and every volume of the Methods in Molecular Biology series. Tested and trusted, comprehensive and reliable, all protocols from the series are indexed in PubMed.

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Molecular model of the mitochondrial genome segregation machinery in Trypanosoma brucei

Cited by 43 publications

References 61 publications

Discovery of new mitochondrial DNA segregation machinery components inTrypanosoma brucei: a comparison of three approaches

Discovery of new mitochondrial DNA segregation machinery components inTrypanosoma brucei: a comparison of three approaches

Dealing with several flagella in the same cell

Airyscan Superresolution Microscopy to Study Trypanosomatid Cell Biology

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