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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...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2022
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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 |
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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 |
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