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Structure paper

TitleStructural insight into TPX2-stimulated microtubule assembly.
Journal, issue, pagesElife, Vol. 6, Year 2017
Publish dateNov 9, 2017
AuthorsRui Zhang / Johanna Roostalu / Thomas Surrey / Eva Nogales /
PubMed AbstractDuring mitosis and meiosis, microtubule (MT) assembly is locally upregulated by the chromatin-dependent Ran-GTP pathway. One of its key targets is the MT-associated spindle assembly factor TPX2. The ...During mitosis and meiosis, microtubule (MT) assembly is locally upregulated by the chromatin-dependent Ran-GTP pathway. One of its key targets is the MT-associated spindle assembly factor TPX2. The molecular mechanism of how TPX2 stimulates MT assembly remains unknown because structural information about the interaction of TPX2 with MTs is lacking. Here, we determine the cryo-electron microscopy structure of a central region of TPX2 bound to the MT surface. TPX2 uses two flexibly linked elements ('ridge' and 'wedge') in a novel interaction mode to simultaneously bind across longitudinal and lateral tubulin interfaces. These MT-interacting elements overlap with the binding site of importins on TPX2. Fluorescence microscopy-based in vitro reconstitution assays reveal that this interaction mode is critical for MT binding and facilitates MT nucleation. Together, our results suggest a molecular mechanism of how the Ran-GTP gradient can regulate TPX2-dependent MT formation.
External linksElife / PubMed:29120325 / PubMed Central
MethodsEM (helical sym.)
Resolution3.3 - 3.6 Å
Structure data

EMDB-7101, PDB-6bjc:
TPX2_mini decorated GMPCPP-microtubule
Method: EM (helical sym.) / Resolution: 3.3 Å

EMDB-7102:
TPX2_micro decorated GMPCPP-microtubule
Method: EM (helical sym.) / Resolution: 3.6 Å

Chemicals

ChemComp-GTP:
GUANOSINE-5'-TRIPHOSPHATE / GTP, energy-carrying molecule*YM

ChemComp-MG:
Unknown entry

ChemComp-G2P:
PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER / GMP-CPP, energy-carrying molecule analogue*YM

Source
  • Sus scrofa (pig)
  • homo sapiens (human)
KeywordsCELL CYCLE / mitosis / GanGTP / nucleation

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