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TitleChanges in seam number and location induce holes within microtubules assembled from porcine brain tubulin and in egg cytoplasmic extracts.
Journal, issue, pagesElife, Vol. 11, Year 2022
Publish dateDec 12, 2022
AuthorsCharlotte Guyomar / Clément Bousquet / Siou Ku / John M Heumann / Gabriel Guilloux / Natacha Gaillard / Claire Heichette / Laurence Duchesne / Michel O Steinmetz / Romain Gibeaux / Denis Chrétien /
PubMed AbstractMicrotubules 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 ...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 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.
External linksElife / PubMed:36503602 / PubMed Central
MethodsEM (tomography) / EM (subtomogram averaging)
Resolution24.1 - 48.5 Å
Structure data

EMDB-15732: Microtubules assembled in Xenopus egg cytoplasmic extract and decorated with kinesin-motor domains
Method: EM (tomography)

EMDB-15733: Microtubule decorated with kinesin-motor domains, 13 protofilaments, 3-start helix, 1 seam
Method: EM (subtomogram averaging) / Resolution: 43.3 Å

EMDB-15734: Microtubule decorated with kinesin-motor domain, 13 protofilaments, 3-start helix, transition from 5 to 3 seams
Method: EM (tomography)

EMDB-15735: Microtubule decorated with kinesin-motor domains, 14 protofilaments, 3-start helix, 1 seam
Method: EM (subtomogram averaging) / Resolution: 25.0 Å

EMDB-15736: Microtubule decorated with kinesin-motor domains; 13 protofilaments, 3-start helix, 3 seams, 2 abnormal protofilaments
Method: EM (subtomogram averaging) / Resolution: 24.1 Å

EMDB-15737: Microtubule decorated with kinesin-motor domains; 13 protofilaments, 3-start helix, 5 seams
Method: EM (subtomogram averaging) / Resolution: 30.7 Å

EMDB-15738: Microtubule decorated with kinesin-motor domains; 13 protofilaments, 3-start helix, 3 seams, 2 abnormal protofilaments
Method: EM (subtomogram averaging) / Resolution: 46.7 Å

EMDB-15739: Microtubule decorated with kinesin-motor domains; 13 protofilaments, 3-start helix, 3 seams
Method: EM (subtomogram averaging) / Resolution: 33.3 Å

EMDB-15740: Microtubule decorated with kinesin-motor domains, 13 protofilaments, 3-start helix, 3 seams
Method: EM (subtomogram averaging) / Resolution: 43.5 Å

EMDB-15741: Microtubule decorated with kinesin-motor domains, 13 protofilaments, 3-start helix, 1 seam
Method: EM (subtomogram averaging) / Resolution: 44.1 Å

EMDB-15742: Microtubule decorated with kinesin-motor 3, 13 protofilaments, 3-start helix, transition from 3 to 1 seams
Method: EM (tomography)

EMDB-15743: Microtubule decorated with kinesin-motor domains, 13 protofilaments, 3-start helix, 1 seam, fully embedded in ice
Method: EM (subtomogram averaging) / Resolution: 48.5 Å

EMDB-15744: Microtubule decorated with kinesin-motor domains, 13 protofilaments, 3-start helix, 0 seam, 1 abnormal protofilament
Method: EM (subtomogram averaging) / Resolution: 45.2 Å

EMDB-15745: Microtubule decorated with kinesin-motor domains, 13 protofilaments, 3-start helix, 1 seam
Method: EM (subtomogram averaging) / Resolution: 45.9 Å

EMDB-15746: Microtubule decorated with kinesin-motor domains, 12 protofilaments, 2-start helix, 2 seams
Method: EM (subtomogram averaging) / Resolution: 45.0 Å

EMDB-15747: Microtubule decorated with kinesin-motor domains, 12 protofilaments, 3-start helix, 1 seam
Method: EM (subtomogram averaging) / Resolution: 44.4 Å

EMDB-15748: Microtubule decorated with kinesin-motor domains, 13 protofilaments, 4-start helix, 0 seam
Method: EM (subtomogram averaging) / Resolution: 27.7 Å

EMDB-15749: Microtubule decorated with kinesin-motor domains, 14 protofilaments, 3-start helix, 1 seam
Method: EM (subtomogram averaging) / Resolution: 27.9 Å

EMDB-15750: Microtubule decorated with kinesin-motor domains, fully embedded in ice
Method: EM (tomography)

EMDB-15751: Microtubule decorated with kinesin-motor domains, 13 protofilaments, 3-start helix, 1 seam, in interaction with the air-water interface
Method: EM (subtomogram averaging) / Resolution: 40.0 Å

EMDB-15752: Microtubule decorated with kinesin-motor domains, interacting with the air-water interface
Method: EM (tomography)

Source
  • Xenopus laevis (African clawed frog)
  • Homo sapiens (human)
  • Sus scrofa domesticus (domestic pig)

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