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A Role for the Membrane in Regulating Chlamydomonas Flagellar Length

Flagellar assembly requires coordination between the assembly of axonemal proteins and the assembly of the flagellar membrane and membrane proteins. Fully grown steady-state Chlamydomonas flagella release flagellar vesicles from their tips and failure to resupply membrane should affect flagellar len...

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Detalles Bibliográficos
Autor principal: Dentler, William
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3554728/
https://www.ncbi.nlm.nih.gov/pubmed/23359798
http://dx.doi.org/10.1371/journal.pone.0053366
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author Dentler, William
author_facet Dentler, William
author_sort Dentler, William
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description Flagellar assembly requires coordination between the assembly of axonemal proteins and the assembly of the flagellar membrane and membrane proteins. Fully grown steady-state Chlamydomonas flagella release flagellar vesicles from their tips and failure to resupply membrane should affect flagellar length. To study vesicle release, plasma and flagellar membrane surface proteins were vectorially pulse-labeled and flagella and vesicles were analyzed for biotinylated proteins. Based on the quantity of biotinylated proteins in purified vesicles, steady-state flagella appeared to shed a minimum of 16% of their surface membrane per hour, equivalent to a complete flagellar membrane being released every 6 hrs or less. Brefeldin-A destroyed Chlamydomonas Golgi, inhibited the secretory pathway, inhibited flagellar regeneration, and induced full-length flagella to disassemble within 6 hrs, consistent with flagellar disassembly being induced by a failure to resupply membrane. In contrast to membrane lipids, a pool of biotinylatable membrane proteins was identified that was sufficient to resupply flagella as they released vesicles for 6 hrs in the absence of protein synthesis and to support one and nearly two regenerations of flagella following amputation. These studies reveal the importance of the secretory pathway to assemble and maintain full-length flagella.
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spelling pubmed-35547282013-01-28 A Role for the Membrane in Regulating Chlamydomonas Flagellar Length Dentler, William PLoS One Research Article Flagellar assembly requires coordination between the assembly of axonemal proteins and the assembly of the flagellar membrane and membrane proteins. Fully grown steady-state Chlamydomonas flagella release flagellar vesicles from their tips and failure to resupply membrane should affect flagellar length. To study vesicle release, plasma and flagellar membrane surface proteins were vectorially pulse-labeled and flagella and vesicles were analyzed for biotinylated proteins. Based on the quantity of biotinylated proteins in purified vesicles, steady-state flagella appeared to shed a minimum of 16% of their surface membrane per hour, equivalent to a complete flagellar membrane being released every 6 hrs or less. Brefeldin-A destroyed Chlamydomonas Golgi, inhibited the secretory pathway, inhibited flagellar regeneration, and induced full-length flagella to disassemble within 6 hrs, consistent with flagellar disassembly being induced by a failure to resupply membrane. In contrast to membrane lipids, a pool of biotinylatable membrane proteins was identified that was sufficient to resupply flagella as they released vesicles for 6 hrs in the absence of protein synthesis and to support one and nearly two regenerations of flagella following amputation. These studies reveal the importance of the secretory pathway to assemble and maintain full-length flagella. Public Library of Science 2013-01-24 /pmc/articles/PMC3554728/ /pubmed/23359798 http://dx.doi.org/10.1371/journal.pone.0053366 Text en © 2013 William Dentler http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Dentler, William
A Role for the Membrane in Regulating Chlamydomonas Flagellar Length
title A Role for the Membrane in Regulating Chlamydomonas Flagellar Length
title_full A Role for the Membrane in Regulating Chlamydomonas Flagellar Length
title_fullStr A Role for the Membrane in Regulating Chlamydomonas Flagellar Length
title_full_unstemmed A Role for the Membrane in Regulating Chlamydomonas Flagellar Length
title_short A Role for the Membrane in Regulating Chlamydomonas Flagellar Length
title_sort role for the membrane in regulating chlamydomonas flagellar length
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3554728/
https://www.ncbi.nlm.nih.gov/pubmed/23359798
http://dx.doi.org/10.1371/journal.pone.0053366
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