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In vivo analysis of outer arm dynein transport reveals cargo-specific intraflagellar transport properties

Outer dynein arms (ODAs) are multiprotein complexes that drive flagellar beating. Based on genetic and biochemical analyses, ODAs preassemble in the cell body and then move into the flagellum by intraflagellar transport (IFT). To study ODA transport in vivo, we expressed the essential intermediate c...

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Autores principales: Dai, Jin, Barbieri, Francesco, Mitchell, David R., Lechtreck, Karl F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6254574/
https://www.ncbi.nlm.nih.gov/pubmed/30133350
http://dx.doi.org/10.1091/mbc.E18-05-0291
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author Dai, Jin
Barbieri, Francesco
Mitchell, David R.
Lechtreck, Karl F.
author_facet Dai, Jin
Barbieri, Francesco
Mitchell, David R.
Lechtreck, Karl F.
author_sort Dai, Jin
collection PubMed
description Outer dynein arms (ODAs) are multiprotein complexes that drive flagellar beating. Based on genetic and biochemical analyses, ODAs preassemble in the cell body and then move into the flagellum by intraflagellar transport (IFT). To study ODA transport in vivo, we expressed the essential intermediate chain 2 tagged with mNeonGreen (IC2-NG) to rescue the corresponding Chlamydomonas reinhardtii mutant oda6. IC2-NG moved by IFT; the transport was of low processivity and increased in frequency during flagellar growth. As expected, IFT of IC2-NG was diminished in oda16, lacking an ODA-specific IFT adapter, and in ift46 IFT46ΔN lacking the ODA16-interacting portion of IFT46. IFT loading appears to involve ODA16-dependent recruitment of ODAs to basal bodies followed by handover to IFT. Upon unloading from IFT, ODAs rapidly docked to the axoneme. Transient docking still occurred in the docking complex mutant oda3 indicating that the docking complex stabilizes rather than initiates ODA–microtubule interactions. In full-length flagella, ODAs continued to enter and move inside cilia by short-term bidirectional IFT and diffusion and the newly imported complexes frequently replaced axoneme-bound ODAs. We propose that the low processivity of ODA-IFT contributes to flagellar maintenance by ensuring the availability of replacement ODAs along the length of flagella.
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spelling pubmed-62545742019-02-04 In vivo analysis of outer arm dynein transport reveals cargo-specific intraflagellar transport properties Dai, Jin Barbieri, Francesco Mitchell, David R. Lechtreck, Karl F. Mol Biol Cell Articles Outer dynein arms (ODAs) are multiprotein complexes that drive flagellar beating. Based on genetic and biochemical analyses, ODAs preassemble in the cell body and then move into the flagellum by intraflagellar transport (IFT). To study ODA transport in vivo, we expressed the essential intermediate chain 2 tagged with mNeonGreen (IC2-NG) to rescue the corresponding Chlamydomonas reinhardtii mutant oda6. IC2-NG moved by IFT; the transport was of low processivity and increased in frequency during flagellar growth. As expected, IFT of IC2-NG was diminished in oda16, lacking an ODA-specific IFT adapter, and in ift46 IFT46ΔN lacking the ODA16-interacting portion of IFT46. IFT loading appears to involve ODA16-dependent recruitment of ODAs to basal bodies followed by handover to IFT. Upon unloading from IFT, ODAs rapidly docked to the axoneme. Transient docking still occurred in the docking complex mutant oda3 indicating that the docking complex stabilizes rather than initiates ODA–microtubule interactions. In full-length flagella, ODAs continued to enter and move inside cilia by short-term bidirectional IFT and diffusion and the newly imported complexes frequently replaced axoneme-bound ODAs. We propose that the low processivity of ODA-IFT contributes to flagellar maintenance by ensuring the availability of replacement ODAs along the length of flagella. The American Society for Cell Biology 2018-10-15 /pmc/articles/PMC6254574/ /pubmed/30133350 http://dx.doi.org/10.1091/mbc.E18-05-0291 Text en © 2018 Dai et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. http://creativecommons.org/licenses/by-nc-sa/3.0 This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License.
spellingShingle Articles
Dai, Jin
Barbieri, Francesco
Mitchell, David R.
Lechtreck, Karl F.
In vivo analysis of outer arm dynein transport reveals cargo-specific intraflagellar transport properties
title In vivo analysis of outer arm dynein transport reveals cargo-specific intraflagellar transport properties
title_full In vivo analysis of outer arm dynein transport reveals cargo-specific intraflagellar transport properties
title_fullStr In vivo analysis of outer arm dynein transport reveals cargo-specific intraflagellar transport properties
title_full_unstemmed In vivo analysis of outer arm dynein transport reveals cargo-specific intraflagellar transport properties
title_short In vivo analysis of outer arm dynein transport reveals cargo-specific intraflagellar transport properties
title_sort in vivo analysis of outer arm dynein transport reveals cargo-specific intraflagellar transport properties
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6254574/
https://www.ncbi.nlm.nih.gov/pubmed/30133350
http://dx.doi.org/10.1091/mbc.E18-05-0291
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