Cargando…

Changes in seam number and location induce holes within microtubules assembled from porcine brain tubulin and in Xenopus egg cytoplasmic extracts

Microtubules are tubes of about 25 nm in diameter that are critically involved in a variety of cellular functions, including motility, compartmentalization, and division. They are considered as pseudo-helical polymers whose constituent αβ-tubulin heterodimers share lateral homotypic interactions, ex...

Descripción completa

Detalles Bibliográficos
Autores principales: Guyomar, Charlotte, Bousquet, Clément, Ku, Siou, Heumann, John M, Guilloux, Gabriel, Gaillard, Natacha, Heichette, Claire, Duchesne, Laurence, Steinmetz, Michel O, Gibeaux, Romain, Chrétien, Denis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9788831/
https://www.ncbi.nlm.nih.gov/pubmed/36503602
http://dx.doi.org/10.7554/eLife.83021
_version_ 1784858841533382656
author Guyomar, Charlotte
Bousquet, Clément
Ku, Siou
Heumann, John M
Guilloux, Gabriel
Gaillard, Natacha
Heichette, Claire
Duchesne, Laurence
Steinmetz, Michel O
Gibeaux, Romain
Chrétien, Denis
author_facet Guyomar, Charlotte
Bousquet, Clément
Ku, Siou
Heumann, John M
Guilloux, Gabriel
Gaillard, Natacha
Heichette, Claire
Duchesne, Laurence
Steinmetz, Michel O
Gibeaux, Romain
Chrétien, Denis
author_sort Guyomar, Charlotte
collection PubMed
description Microtubules are tubes of about 25 nm in diameter that are critically involved in a variety of cellular functions, including motility, compartmentalization, and division. They are considered as pseudo-helical polymers whose constituent αβ-tubulin heterodimers share lateral homotypic interactions, except at one unique region called the seam. Here, we used a segmented sub-tomogram averaging strategy to reassess this paradigm and analyze the organization of the αβ-tubulin heterodimers in microtubules assembled from purified porcine brain tubulin in the presence of GTP and GMPCPP, and in Xenopus egg cytoplasmic extracts. We find that in almost all conditions, microtubules incorporate variable protofilament and/or tubulin subunit helical-start numbers, as well as variable numbers of seams. Strikingly, the seam number and location vary along individual microtubules, generating holes of one to a few subunits in size within their lattices. Together, our results reveal that the formation of mixed and discontinuous microtubule lattices is an intrinsic property of tubulin that requires the formation of unique lateral interactions without longitudinal ones. They further suggest that microtubule assembly is tightly regulated in a cytoplasmic environment.
format Online
Article
Text
id pubmed-9788831
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher eLife Sciences Publications, Ltd
record_format MEDLINE/PubMed
spelling pubmed-97888312022-12-24 Changes in seam number and location induce holes within microtubules assembled from porcine brain tubulin and in Xenopus egg cytoplasmic extracts Guyomar, Charlotte Bousquet, Clément Ku, Siou Heumann, John M Guilloux, Gabriel Gaillard, Natacha Heichette, Claire Duchesne, Laurence Steinmetz, Michel O Gibeaux, Romain Chrétien, Denis eLife Cell Biology Microtubules are tubes of about 25 nm in diameter that are critically involved in a variety of cellular functions, including motility, compartmentalization, and division. They are considered as pseudo-helical polymers whose constituent αβ-tubulin heterodimers share lateral homotypic interactions, except at one unique region called the seam. Here, we used a segmented sub-tomogram averaging strategy to reassess this paradigm and analyze the organization of the αβ-tubulin heterodimers in microtubules assembled from purified porcine brain tubulin in the presence of GTP and GMPCPP, and in Xenopus egg cytoplasmic extracts. We find that in almost all conditions, microtubules incorporate variable protofilament and/or tubulin subunit helical-start numbers, as well as variable numbers of seams. Strikingly, the seam number and location vary along individual microtubules, generating holes of one to a few subunits in size within their lattices. Together, our results reveal that the formation of mixed and discontinuous microtubule lattices is an intrinsic property of tubulin that requires the formation of unique lateral interactions without longitudinal ones. They further suggest that microtubule assembly is tightly regulated in a cytoplasmic environment. eLife Sciences Publications, Ltd 2022-12-12 /pmc/articles/PMC9788831/ /pubmed/36503602 http://dx.doi.org/10.7554/eLife.83021 Text en © 2022, Guyomar et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Cell Biology
Guyomar, Charlotte
Bousquet, Clément
Ku, Siou
Heumann, John M
Guilloux, Gabriel
Gaillard, Natacha
Heichette, Claire
Duchesne, Laurence
Steinmetz, Michel O
Gibeaux, Romain
Chrétien, Denis
Changes in seam number and location induce holes within microtubules assembled from porcine brain tubulin and in Xenopus egg cytoplasmic extracts
title Changes in seam number and location induce holes within microtubules assembled from porcine brain tubulin and in Xenopus egg cytoplasmic extracts
title_full Changes in seam number and location induce holes within microtubules assembled from porcine brain tubulin and in Xenopus egg cytoplasmic extracts
title_fullStr Changes in seam number and location induce holes within microtubules assembled from porcine brain tubulin and in Xenopus egg cytoplasmic extracts
title_full_unstemmed Changes in seam number and location induce holes within microtubules assembled from porcine brain tubulin and in Xenopus egg cytoplasmic extracts
title_short Changes in seam number and location induce holes within microtubules assembled from porcine brain tubulin and in Xenopus egg cytoplasmic extracts
title_sort changes in seam number and location induce holes within microtubules assembled from porcine brain tubulin and in xenopus egg cytoplasmic extracts
topic Cell Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9788831/
https://www.ncbi.nlm.nih.gov/pubmed/36503602
http://dx.doi.org/10.7554/eLife.83021
work_keys_str_mv AT guyomarcharlotte changesinseamnumberandlocationinduceholeswithinmicrotubulesassembledfromporcinebraintubulinandinxenopuseggcytoplasmicextracts
AT bousquetclement changesinseamnumberandlocationinduceholeswithinmicrotubulesassembledfromporcinebraintubulinandinxenopuseggcytoplasmicextracts
AT kusiou changesinseamnumberandlocationinduceholeswithinmicrotubulesassembledfromporcinebraintubulinandinxenopuseggcytoplasmicextracts
AT heumannjohnm changesinseamnumberandlocationinduceholeswithinmicrotubulesassembledfromporcinebraintubulinandinxenopuseggcytoplasmicextracts
AT guillouxgabriel changesinseamnumberandlocationinduceholeswithinmicrotubulesassembledfromporcinebraintubulinandinxenopuseggcytoplasmicextracts
AT gaillardnatacha changesinseamnumberandlocationinduceholeswithinmicrotubulesassembledfromporcinebraintubulinandinxenopuseggcytoplasmicextracts
AT heichetteclaire changesinseamnumberandlocationinduceholeswithinmicrotubulesassembledfromporcinebraintubulinandinxenopuseggcytoplasmicextracts
AT duchesnelaurence changesinseamnumberandlocationinduceholeswithinmicrotubulesassembledfromporcinebraintubulinandinxenopuseggcytoplasmicextracts
AT steinmetzmichelo changesinseamnumberandlocationinduceholeswithinmicrotubulesassembledfromporcinebraintubulinandinxenopuseggcytoplasmicextracts
AT gibeauxromain changesinseamnumberandlocationinduceholeswithinmicrotubulesassembledfromporcinebraintubulinandinxenopuseggcytoplasmicextracts
AT chretiendenis changesinseamnumberandlocationinduceholeswithinmicrotubulesassembledfromporcinebraintubulinandinxenopuseggcytoplasmicextracts