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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-...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2018
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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. |
format | Online Article Text |
id | pubmed-5931796 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
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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