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dc.contributor.authorCraft Van De Weghe, Julie
dc.contributor.authorHarris, J. Aaron
dc.contributor.authorKubo, Tomohiro
dc.contributor.authorWitman, George B.
dc.contributor.authorLechtreck, Karl F.
dc.date2022-08-11T08:10:49.000
dc.date.accessioned2022-08-23T17:21:09Z
dc.date.available2022-08-23T17:21:09Z
dc.date.issued2020-09-11
dc.date.submitted2020-10-21
dc.identifier.citation<p>Craft Van De Weghe J, Harris JA, Kubo T, Witman GB, Lechtreck KF. Diffusion rather than intraflagellar transport likely provides most of the tubulin required for axonemal assembly in <em>Chlamydomonas</em>. J Cell Sci. 2020 Sep 11;133(17):jcs249805. doi: 10.1242/jcs.249805. PMID: 32801124; PMCID: PMC7502601. <a href="https://doi.org/10.1242/jcs.249805">Link to article on publisher's site</a></p>
dc.identifier.issn0021-9533 (Linking)
dc.identifier.doi10.1242/jcs.249805
dc.identifier.pmid32801124
dc.identifier.urihttp://hdl.handle.net/20.500.14038/48458
dc.description.abstractTubulin enters the cilium by diffusion and motor-based intraflagellar transport (IFT). However, the respective contribution of each route in providing tubulin for axonemal assembly remains unknown. Using Chlamydomonas, we attenuated IFT-based tubulin transport of GFP-beta-tubulin by altering the IFT74N-IFT81N tubulin-binding module and the C-terminal E-hook of tubulin. E-hook-deficient GFP-beta-tubulin was incorporated into the axonemal microtubules, but its transport frequency by IFT was reduced by approximately 90% in control cells and essentially abolished when the tubulin-binding site of IFT81 was incapacitated. Despite the strong reduction in IFT, the proportion of E-hook-deficient GFP-beta-tubulin in the axoneme was only moderately reduced. In vivo imaging showed more GFP-beta-tubulin particles entering cilia by diffusion than by IFT. Extrapolated to endogenous tubulin, the data indicate that diffusion provides most of the tubulin required for axonemal assembly. We propose that IFT of tubulin is nevertheless needed for ciliogenesis, because it augments the tubulin pool supplied to the ciliary tip by diffusion, thus ensuring that free tubulin there is maintained at the critical concentration for plus-end microtubule assembly during rapid ciliary growth.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=32801124&dopt=Abstract">Link to Article in PubMed</a></p>
dc.rights© 2020. Published by The Company of Biologists Ltd. Publisher PDF posted after 12 months as allowed by publisher's author rights policy at https://www.biologists.com/user-licence-1-1/.
dc.subjectFlagella
dc.subjectCilia
dc.subjectMicrotubule
dc.subjectAxoneme
dc.subjectCalponin homology domain
dc.subjectIFT81IFT74
dc.subjectDiffusion
dc.subjectAlgae
dc.subjectAmino Acids, Peptides, and Proteins
dc.subjectCell Biology
dc.subjectCellular and Molecular Physiology
dc.subjectDevelopmental Biology
dc.titleDiffusion rather than intraflagellar transport likely provides most of the tubulin required for axonemal assembly in Chlamydomonas
dc.typeJournal Article
dc.source.journaltitleJournal of cell science
dc.source.volume133
dc.source.issue17
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=1575&amp;context=radiology_pubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/radiology_pubs/563
dc.legacy.embargo2021-09-11T00:00:00-07:00
dc.identifier.contextkey19898211
refterms.dateFOA2022-08-23T17:21:09Z
html.description.abstract<p>Tubulin enters the cilium by diffusion and motor-based intraflagellar transport (IFT). However, the respective contribution of each route in providing tubulin for axonemal assembly remains unknown. Using Chlamydomonas, we attenuated IFT-based tubulin transport of GFP-beta-tubulin by altering the IFT74N-IFT81N tubulin-binding module and the C-terminal E-hook of tubulin. E-hook-deficient GFP-beta-tubulin was incorporated into the axonemal microtubules, but its transport frequency by IFT was reduced by approximately 90% in control cells and essentially abolished when the tubulin-binding site of IFT81 was incapacitated. Despite the strong reduction in IFT, the proportion of E-hook-deficient GFP-beta-tubulin in the axoneme was only moderately reduced. In vivo imaging showed more GFP-beta-tubulin particles entering cilia by diffusion than by IFT. Extrapolated to endogenous tubulin, the data indicate that diffusion provides most of the tubulin required for axonemal assembly. We propose that IFT of tubulin is nevertheless needed for ciliogenesis, because it augments the tubulin pool supplied to the ciliary tip by diffusion, thus ensuring that free tubulin there is maintained at the critical concentration for plus-end microtubule assembly during rapid ciliary growth.</p>
dc.identifier.submissionpathradiology_pubs/563
dc.contributor.departmentWitman Lab
dc.contributor.departmentDepartment of Radiology
dc.source.pagesjcs249805


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