A novel approach to tagging tubulin reveals microtubule assembly dynamics of the axoneme in Trypanosoma brucei

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68378050%3A_____%2F25%3A00641324" target="_blank" >RIV/68378050:_____/25:00641324 - isvavai.cz</a>

Výsledek na webu

<a href="https://doi.org/10.1242/jcs.264145" target="_blank" >https://doi.org/10.1242/jcs.264145</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1242/jcs.264145" target="_blank" >10.1242/jcs.264145</a>

Jazyk výsledku

angličtina

Název v původním jazyce

A novel approach to tagging tubulin reveals microtubule assembly dynamics of the axoneme in Trypanosoma brucei

Popis výsledku v původním jazyce

The protozoan parasite Trypanosoma brucei assembles a new flagellum while maintaining the existing one in the same cell. Our group has previously proposed a model where the mature flagellum is locked after construction to full length. To test this hypothesis directly, we monitored flagellum assembly dynamics through inducible expression of tubulin marked with an intragenic tag. We found that addition of new tubulin occurs at the distal flagellum tip at a linear rate and is indeed restricted to the new flagellum in bi-flagellated cells. Depleting the locking protein CEP164C prior to induction resulted in simultaneous integration of new tubulin in both flagella. This is direct evidence that trypanosomes avoid competition between the two flagella by allowing tubulin incorporation only in the new organelle. However, by tracing flagella over several cell cycles we also found that flagella do not remain locked forever. An orthogonal approach with HaloTag-tagged radial spoke protein 4/6 (GeneID Tb927.11.4480) supported these findings. Given that flagellum length in trypanosomes is stable, this indicates regular events of transient disassembly, followed by assembly, at the distal tip.

Název v anglickém jazyce

A novel approach to tagging tubulin reveals microtubule assembly dynamics of the axoneme in Trypanosoma brucei

Popis výsledku anglicky

The protozoan parasite Trypanosoma brucei assembles a new flagellum while maintaining the existing one in the same cell. Our group has previously proposed a model where the mature flagellum is locked after construction to full length. To test this hypothesis directly, we monitored flagellum assembly dynamics through inducible expression of tubulin marked with an intragenic tag. We found that addition of new tubulin occurs at the distal flagellum tip at a linear rate and is indeed restricted to the new flagellum in bi-flagellated cells. Depleting the locking protein CEP164C prior to induction resulted in simultaneous integration of new tubulin in both flagella. This is direct evidence that trypanosomes avoid competition between the two flagella by allowing tubulin incorporation only in the new organelle. However, by tracing flagella over several cell cycles we also found that flagella do not remain locked forever. An orthogonal approach with HaloTag-tagged radial spoke protein 4/6 (GeneID Tb927.11.4480) supported these findings. Given that flagellum length in trypanosomes is stable, this indicates regular events of transient disassembly, followed by assembly, at the distal tip.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/GA23-07370S" target="_blank" >GA23-07370S: Nutriční stress jako spouštěč buněčné diferenciace procyclické formy Trypanosoma brucei</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Cell Science

ISSN

0021-9533

e-ISSN

1477-9137

Svazek periodika

138

Číslo periodika v rámci svazku

20

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

18

Strana od-do

jcs264145

Kód UT WoS článku

001604761900005

EID výsledku v databázi Scopus

2-s2.0-105010719822