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Crystal structure of intraflagellar transport protein 80 reveals a homo-dimer required for ciliogenesis

Oligomeric assemblies of intraflagellar transport (IFT) particles build cilia through sequential recruitment and transport of ciliary cargo proteins within cilia. Here we present the 1.8 Å resolution crystal structure of the Chlamydomonas IFT-B protein IFT80, which reveals the architecture of two N-...

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Autores principales: Taschner, Michael, Lorentzen, Anna, Mourão, André, Collins, Toby, Freke, Grace M, Moulding, Dale, Basquin, Jerome, Jenkins, Dagan, Lorentzen, Esben
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5931796/
https://www.ncbi.nlm.nih.gov/pubmed/29658880
http://dx.doi.org/10.7554/eLife.33067
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author Taschner, Michael
Lorentzen, Anna
Mourão, André
Collins, Toby
Freke, Grace M
Moulding, Dale
Basquin, Jerome
Jenkins, Dagan
Lorentzen, Esben
author_facet Taschner, Michael
Lorentzen, Anna
Mourão, André
Collins, Toby
Freke, Grace M
Moulding, Dale
Basquin, Jerome
Jenkins, Dagan
Lorentzen, Esben
author_sort Taschner, Michael
collection PubMed
description Oligomeric assemblies of intraflagellar transport (IFT) particles build cilia through sequential recruitment and transport of ciliary cargo proteins within cilia. Here we present the 1.8 Å resolution crystal structure of the Chlamydomonas IFT-B protein IFT80, which reveals the architecture of two N-terminal β-propellers followed by an α-helical extension. The N-terminal β-propeller tethers IFT80 to the IFT-B complex via IFT38 whereas the second β-propeller and the C-terminal α-helical extension result in IFT80 homo-dimerization. Using CRISPR/Cas to create biallelic Ift80 frameshift mutations in IMCD3 mouse cells, we demonstrate that IFT80 is absolutely required for ciliogenesis. Structural mapping and rescue experiments reveal that human disease-causing missense mutations do not cluster within IFT80 and form functional IFT particles. Unlike missense mutant forms of IFT80, deletion of the C-terminal dimerization domain prevented rescue of ciliogenesis. Taken together our results may provide a first insight into higher order IFT complex formation likely required for IFT train formation.
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spelling pubmed-59317962018-05-03 Crystal structure of intraflagellar transport protein 80 reveals a homo-dimer required for ciliogenesis Taschner, Michael Lorentzen, Anna Mourão, André Collins, Toby Freke, Grace M Moulding, Dale Basquin, Jerome Jenkins, Dagan Lorentzen, Esben eLife Cell Biology Oligomeric assemblies of intraflagellar transport (IFT) particles build cilia through sequential recruitment and transport of ciliary cargo proteins within cilia. Here we present the 1.8 Å resolution crystal structure of the Chlamydomonas IFT-B protein IFT80, which reveals the architecture of two N-terminal β-propellers followed by an α-helical extension. The N-terminal β-propeller tethers IFT80 to the IFT-B complex via IFT38 whereas the second β-propeller and the C-terminal α-helical extension result in IFT80 homo-dimerization. Using CRISPR/Cas to create biallelic Ift80 frameshift mutations in IMCD3 mouse cells, we demonstrate that IFT80 is absolutely required for ciliogenesis. Structural mapping and rescue experiments reveal that human disease-causing missense mutations do not cluster within IFT80 and form functional IFT particles. Unlike missense mutant forms of IFT80, deletion of the C-terminal dimerization domain prevented rescue of ciliogenesis. Taken together our results may provide a first insight into higher order IFT complex formation likely required for IFT train formation. eLife Sciences Publications, Ltd 2018-04-16 /pmc/articles/PMC5931796/ /pubmed/29658880 http://dx.doi.org/10.7554/eLife.33067 Text en © 2018, Taschner et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Cell Biology
Taschner, Michael
Lorentzen, Anna
Mourão, André
Collins, Toby
Freke, Grace M
Moulding, Dale
Basquin, Jerome
Jenkins, Dagan
Lorentzen, Esben
Crystal structure of intraflagellar transport protein 80 reveals a homo-dimer required for ciliogenesis
title Crystal structure of intraflagellar transport protein 80 reveals a homo-dimer required for ciliogenesis
title_full Crystal structure of intraflagellar transport protein 80 reveals a homo-dimer required for ciliogenesis
title_fullStr Crystal structure of intraflagellar transport protein 80 reveals a homo-dimer required for ciliogenesis
title_full_unstemmed Crystal structure of intraflagellar transport protein 80 reveals a homo-dimer required for ciliogenesis
title_short Crystal structure of intraflagellar transport protein 80 reveals a homo-dimer required for ciliogenesis
title_sort crystal structure of intraflagellar transport protein 80 reveals a homo-dimer required for ciliogenesis
topic Cell Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5931796/
https://www.ncbi.nlm.nih.gov/pubmed/29658880
http://dx.doi.org/10.7554/eLife.33067
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