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Structure paper

TitleThe role of tubulin-tubulin lattice contacts in the mechanism of microtubule dynamic instability.
Journal, issue, pagesNat Struct Mol Biol, Vol. 25, Issue 7, Page 607-615, Year 2018
Publish dateJul 2, 2018
AuthorsSzymon W Manka / Carolyn A Moores /
PubMed AbstractMicrotubules form from longitudinally and laterally assembling tubulin α-β dimers. The assembly induces strain in tubulin, resulting in cycles of microtubule catastrophe and regrowth. This 'dynamic ...Microtubules form from longitudinally and laterally assembling tubulin α-β dimers. The assembly induces strain in tubulin, resulting in cycles of microtubule catastrophe and regrowth. This 'dynamic instability' is governed by GTP hydrolysis that renders the microtubule lattice unstable, but it is unclear how. We used a human microtubule nucleating and stabilizing neuronal protein, doublecortin, and high-resolution cryo-EM to capture tubulin's elusive hydrolysis intermediate GDP•Pi state, alongside the prehydrolysis analog GMPCPP state and the posthydrolysis GDP state with and without an anticancer drug, Taxol. GTP hydrolysis to GDP•Pi followed by Pi release constitutes two distinct structural transitions, causing unevenly distributed compressions of tubulin dimers, thereby tightening longitudinal and loosening lateral interdimer contacts. We conclude that microtubule catastrophe is triggered because the lateral contacts can no longer counteract the strain energy stored in the lattice, while reinforcement of the longitudinal contacts may support generation of force.
External linksNat Struct Mol Biol / PubMed:29967541 / PubMed Central
MethodsEM (single particle)
Resolution3.8 - 4.4 Å
Structure data

EMDB-3961, PDB-6evw:
Cryo-EM structure of GMPCPP-microtubule co-polymerised with doublecortin
Method: EM (single particle) / Resolution: 4.4 Å

EMDB-3962, PDB-6evx:
Cryo-EM structure of GDP.Pi-microtubule rapidly co-polymerised with doublecortin
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-3963, PDB-6evy:
Cryo-EM structure of GTPgammaS-microtubule co-polymerised with doublecortin
Method: EM (single particle) / Resolution: 4.4 Å

EMDB-3964, PDB-6evz:
Cryo-EM structure of GDP-microtubule co-polymerised with doublecortin
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-3965, PDB-6ew0:
Cryo-EM structure of GDP-microtubule co-polymerised with doublecortin and supplemented with Taxol
Method: EM (single particle) / Resolution: 3.8 Å

Chemicals

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

ChemComp-MG:
Unknown entry

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

ChemComp-GDP:
GUANOSINE-5'-DIPHOSPHATE / GDP, energy-carrying molecule*YM / Guanosine diphosphate

ChemComp-PO4:
PHOSPHATE ION / Phosphate

ChemComp-GSP:
5'-GUANOSINE-DIPHOSPHATE-MONOTHIOPHOSPHATE

ChemComp-TA1:
TAXOL / medication, chemotherapy*YM / Paclitaxel

Source
  • sus sucrofa (pig)
  • Pig (pig)
  • sus scrofa (pig)
KeywordsSTRUCTURAL PROTEIN / microtubule / GTPase / tubulin / Taxol

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