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<p>The 9 + 2 microtubule-based axoneme within motile flagella is well known for its symmetry. However, examples of asymmetric structures and proteins asymmetrically positioned within the 9 + 2 axoneme architecture have been identified. These occur in multiple different organisms, particularly involving the inner or outer dynein arms. Here, we comprehensively analyse conserved proximal-distal asymmetries in the uniflagellate trypanosomatid eukaryotic parasites. Building on the genome-wide localisation screen in <i>Trypanosoma brucei</i> we identify conserved proteins with an analogous asymmetric localisation in the related parasite <i>Leishmania mexicana</i>. Using deletion mutants, we find which are necessary for normal cell swimming, flagellum beat parameters and axoneme ultrastructure. Using combinatorial endogenous fluorescent tagging and deletion, we map co-dependencies for assembly into their normal asymmetric localisation. This revealed 15 proteins, 9 known and 6 novel, with a conserved proximal or distal axoneme-specific localisation. Most are outer dynein arm associated and show that there are multiple classes of proximal-distal asymmetry – one which is dependent on the docking complex. Many of these&#xa0;proteins are necessary for retaining the normal frequency of the tip-to-base symmetric flagellar waveform. Our comprehensive mapping reveals unexpected contributions of proximal-specific axoneme components to the frequency of waveforms initiated distally.</p>

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Proteins with proximal-distal asymmetries in axoneme localisation control flagellum beat frequency

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年4月4日

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  • Cecile Fort,
  • Benjamin J. Walker,
  • Lore Baert,
  • Richard J. Wheeler

摘要

The 9 + 2 microtubule-based axoneme within motile flagella is well known for its symmetry. However, examples of asymmetric structures and proteins asymmetrically positioned within the 9 + 2 axoneme architecture have been identified. These occur in multiple different organisms, particularly involving the inner or outer dynein arms. Here, we comprehensively analyse conserved proximal-distal asymmetries in the uniflagellate trypanosomatid eukaryotic parasites. Building on the genome-wide localisation screen in Trypanosoma brucei we identify conserved proteins with an analogous asymmetric localisation in the related parasite Leishmania mexicana. Using deletion mutants, we find which are necessary for normal cell swimming, flagellum beat parameters and axoneme ultrastructure. Using combinatorial endogenous fluorescent tagging and deletion, we map co-dependencies for assembly into their normal asymmetric localisation. This revealed 15 proteins, 9 known and 6 novel, with a conserved proximal or distal axoneme-specific localisation. Most are outer dynein arm associated and show that there are multiple classes of proximal-distal asymmetry – one which is dependent on the docking complex. Many of these proteins are necessary for retaining the normal frequency of the tip-to-base symmetric flagellar waveform. Our comprehensive mapping reveals unexpected contributions of proximal-specific axoneme components to the frequency of waveforms initiated distally.