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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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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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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 |
collection | PubMed |
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. |
format | Online Article Text |
id | pubmed-3554728 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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