WDR60-mediated dynein-2 loading into cilia powers retrograde IFT and transition zone crossing
| Autor: | Diogo R. M. Rodrigues, Ana Carvalho, Neide Vieira, Tiago J. Dantas, Cármen Vieira, Reto Gassmann, Maria J. G. De-Castro, Carla M. Abreu, Ana R. G. De-Castro |
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| Přispěvatelé: | Instituto de Investigação e Inovação em Saúde |
| Rok vydání: | 2021 |
| Předmět: |
Sensory Receptor Cells
Dynein Green Fluorescent Proteins Motility Development Biology 03 medical and health sciences 0302 clinical medicine Protein Domains Intraflagellar transport Live cell imaging Transition zone Genetics Animals Cilia Ciliary tip Caenorhabditis elegans Caenorhabditis elegans Proteins Cytoskeleton 030304 developmental biology 0303 health sciences Chemistry Cilium Dyneins Cell Biology biology.organism_classification Cell biology Cytoskeletal Proteins Kinetics Flagella Mutation sense organs Humanities 030217 neurology & neurosurgery |
| Zdroj: | Journal of Cell Biology. 221 |
| ISSN: | 1540-8140 0021-9525 |
| Popis: | The dynein-2 motor complex drives retrograde intraflagellar transport (IFT), playing a pivotal role in the assembly and functions of cilia. However, the mechanisms that regulate dynein-2 motility remain poorly understood. Here, we identify the Caenorhabditis elegans WDR60 homologue, WDR-60, and dissect the roles of this intermediate chain using genome editing and live imaging of endogenous dynein-2/IFT components. We find that loss of WDR-60 impairs dynein-2 recruitment to cilia and its incorporation onto anterograde IFT trains, reducing retrograde motor availability at the ciliary tip. Consistent with this, we show that fewer dynein-2 motors power WDR-60-deficient retrograde IFT trains, which move at reduced velocities and fail to exit cilia, accumulating on the distal side of the transition zone. Remarkably, disrupting the transition zone's NPHP module almost fully restores ciliary exit of underpowered retrograde trains in wdr-60 mutants. This work establishes WDR-60 as a major contributor to IFT, and the NPHP module as a roadblock to dynein-2 passage through the transition zone. This work was financed by Fundo Europeu de Desenvolvimento Regional (FEDER) through the COMPETE 2020 Operacional Programme for Competitiveness and Internationalisation (POCI), Portugal 2020, and Portuguese funds through Fundação para a Ciência e a Tecnologia (FCT)/Minist´erio da Ciência, Tecnologia e Ensino Superior in the framework of the project POCI-01-0145-FEDER-029471 (PTDC/BIA-BID/29471/2017) to T.J. Dantas. A.X. Carvalho, R. Gassmann, C.M.C. Abreu, and T.J. Dantas were supported by the FCT: CEECIND/01967/2017, CEECIND/00333/2017, CEECIND/01985/2018, and CEECIND/00771/2017, respectively. N. Vieira is a Junior Researcher under the scope of the FCT Transitional Rule DL57/2016, and her work was supported by a 2016 NARSAD Young Investigator Grant (#24929) from the Brain and Behavior Research Foundation. D.R.M. Rodrigues received a PhD fellowship from FCT (SFRH/BD/143985/2019) and support fromthe Biomedical Sciences PhD program at Instituto de Ciências Biom´edicas Abel Salazar (ICBAS). Some strains were provided by the National Bioresource Project for C. elegans and by the Caenorhabditis Genetics Center (CGC), which is funded by the National Institutes of Health Office of Research Infrastructure Programs (P40 OD010440). |
| Databáze: | OpenAIRE |
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