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TitleCryo-ET of parasites gives subnanometer insight into tubulin-based structures.
Journal, issue, pagesProc Natl Acad Sci U S A, Vol. 119, Issue 6, Year 2022
Publish dateFeb 8, 2022
AuthorsStella Y Sun / Li-Av Segev-Zarko / Muyuan Chen / Grigore D Pintilie / Michael F Schmid / Steven J Ludtke / John C Boothroyd / Wah Chiu /
PubMed AbstractTubulin is a conserved protein that polymerizes into different forms of filamentous structures in , an obligate intracellular parasite in the phylum Apicomplexa. Two key tubulin-containing ...Tubulin is a conserved protein that polymerizes into different forms of filamentous structures in , an obligate intracellular parasite in the phylum Apicomplexa. Two key tubulin-containing cytoskeletal components are subpellicular microtubules (SPMTs) and conoid fibrils (CFs). The SPMTs help maintain shape and gliding motility, while the CFs are implicated in invasion. Here, we use cryogenic electron tomography to determine the molecular structures of the SPMTs and CFs in vitrified intact and detergent-extracted parasites. Subvolume densities from detergent-extracted parasites yielded averaged density maps at subnanometer resolutions, and these were related back to their architecture in situ. An intralumenal spiral lines the interior of the 13-protofilament SPMTs, revealing a preferred orientation of these microtubules relative to the parasite's long axis. Each CF is composed of nine tubulin protofilaments that display a comma-shaped cross-section, plus additional associated components. Conoid protrusion, a crucial step in invasion, is associated with an altered pitch of each CF. The use of basic building blocks of protofilaments and different accessory proteins in one organism illustrates the versatility of tubulin to form two distinct types of assemblies, SPMTs and CFs.
External linksProc Natl Acad Sci U S A / PubMed:35121661 / PubMed Central
MethodsEM (tomography) / EM (subtomogram averaging)
Resolution6.7 - 46.0 Å
Structure data

EMDB-26006:
3D reconstruction of detergent-extract Toxoplasma gondii cells at the apical end
Method: EM (tomography)

EMDB-26007:
Three-dimensional organization of the apical complex in Toxoplasma tachyzoites
Method: EM (tomography)

EMDB-26008:
In situ average map of conoid with PCR from intact Toxoplasma gondii cells
Method: EM (subtomogram averaging) / Resolution: 46.0 Å

EMDB-26009:
The conoid segment from intact Toxoplasma gondii cells
Method: EM (subtomogram averaging) / Resolution: 28.0 Å

EMDB-26010:
The symmetrized subpellicular microtubule map from intact Toxoplasma gondii cells
Method: EM (subtomogram averaging) / Resolution: 26.0 Å

26018

7tnq

EMDB-26018, PDB-7tnq:
The symmetry-released subpellicular microtubule map from detergent-extracted Toxoplasma cells
Method: EM (subtomogram averaging) / Resolution: 8.4 Å

26019

7tns

EMDB-26019, PDB-7tns:
Subpellicular microtubule from detergent-extract Toxoplasma gondii cells
Method: EM (subtomogram averaging) / Resolution: 6.7 Å

26020

7tnt

EMDB-26020, PDB-7tnt:
The tubulin-based conoid from detergent-extract Toxoplasma gondii cells
Method: EM (subtomogram averaging) / Resolution: 9.3 Å

Source
  • toxoplasma gondii (eukaryote)
KeywordsCELL INVASION / parasites / Toxoplasma gondii / cytoskeleton / microtubules / tubulin / conoid

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