PXD059041

PXD059041 is an original dataset announced via ProteomeXchange.

Title	A novel approach to tagging tubulin reveals MT assembly dynamics of the axoneme in Trypanosoma brucei
Description	The protozoan parasite Trypanosoma brucei is a mono-flagellated cell during the G1-phase of its cell cycle. In order to duplicate, it assembles a new flagellum alongside the mature one, in which further elongation is prevented. Our group proposed a model where the mature flagellum is locked after construction to full length (Bertiaux et al. 2018) and access of new building blocks for elongation is exclusive to the new flagellum. To test this hypothesis directly, we developed a tool for the inducible expression of tagged tubulin. Alpha-tubulin that was tagged with an intragenic Ty-1-epitope behaved indistinguishable from untagged tubulin. Its incorporation was monitored after inducible expression, to follow the assembly dynamics of microtubules in the cell body, the mitotic spindle and the flagellum. In this study we present direct evidence that integration of tubulin occurs at the distal flagellum tip at a linear rate and is indeed restricted to the new flagellum in bi-flagellated cells. This is direct evidence that trypanosomes avoid competition between the two flagella by allowing tubulin incorporation only in the new but not the old organelle. However, by tracing flagella over several cell cycles we could also show that mature flagella do not remain locked indefinitely. The restriction is lifted briefly after the bi-flagellated cell has divided and the daughter cell inheriting the old flagellum shows incorporation of newly synthesized building blocks again. It then has to lock again before the cell can assemble a new flagellum. Our findings suggest regular incorporation of tubulin at the tip of previously locked flagella. Since flagellum length in trypanosomes is stable, it is likely that regular dis-assembly needs to occur to prevent a net elongation over time. This evidence was supported with an orthogonal approach, with which we monitored the incorporation of HALO-tagged radial spoke protein 4/6
HostingRepository	PRIDE
AnnounceDate	2025-06-18
AnnouncementXML	Submission_2025-06-18_04:56:21.772.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	MARIETTE MATONDO
SpeciesList	scientific name: Trypanosoma brucei; NCBI TaxID: 5691;
ModificationList	acetylated residue
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2024-12-19 02:26:47	ID requested
⏵ 1	2025-06-18 04:56:22	announced

Dataset with its publication pending

submitter keyword: PTMS, acetylation,Tubbulines

Bastin Philippe
contact affiliation	rypanosome Cell Biology Unit & INSERM U1201 Institut Pasteur, 25, rue du Docteur Roux, 75015 Paris (France)
contact email	philippe.bastin@pasteur.fr
lab head
MARIETTE MATONDO
contact affiliation	Pasteur
contact email	mariette.matondo@pasteur.fr
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD059041
     1. Label: PRIDE project
     2. Name: A novel approach to tagging tubulin reveals MT assembly dynamics of the axoneme in Trypanosoma brucei