TY - JOUR
T1 - High-Resolution microtubule structures reveal the structural transitions in αβ-tubulin upon GTP hydrolysis
AU - Alushin, Gregory M.
AU - Lander, Gabriel C.
AU - Kellogg, Elizabeth H.
AU - Zhang, Rui
AU - Baker, David
AU - Nogales, Eva
N1 - Funding Information:
We thank Patricia Grob and Tom Houweling for electron microscopy and computer support, respectively, Tom Goddard for help with UCSF Chimera, Yifan Song and Frank DiMaio for making code available for use prior to publication, and Stuart Howes for assistance with kinesin purification. We are grateful to Robert Glaeser for assistance with data collection and many discussions. The kinesin expression construct was the gift of Erik Jonsson and Ron Vale. This work was funded by the National Institute of General Medical Sciences (GM051487) (to E.N.) and a Damon Runyon Cancer Research Foundation fellowship (DRG 2055-10) (to G.C.L.). E.N. and D.B. are Howard Hughes Medical Institute Investigators.
PY - 2014/5/22
Y1 - 2014/5/22
N2 - Dynamic instability, the stochastic switching between growth and shrinkage, is essential for microtubule function. This behavior is driven by GTP hydrolysis in the microtubule lattice and is inhibited by anticancer agents like Taxol. We provide insight into the mechanism of dynamic instability, based on high-resolution cryo-EM structures (4.7-5.6 Å) of dynamic microtubules and microtubules stabilized by GMPCPP or Taxol. We infer that hydrolysis leads to a compaction around the E-site nucleotide at longitudinal interfaces, as well as movement of the α-tubulin intermediate domain and H7 helix. Displacement of the C-terminal helices in both α- and β-tubulin subunits suggests an effect on interactions with binding partners that contact this region. Taxol inhibits most of these conformational changes, allosterically inducing a GMPCPP-like state. Lateral interactions are similar in all conditions we examined, suggesting that microtubule lattice stability is primarily modulated at longitudinal interfaces.
AB - Dynamic instability, the stochastic switching between growth and shrinkage, is essential for microtubule function. This behavior is driven by GTP hydrolysis in the microtubule lattice and is inhibited by anticancer agents like Taxol. We provide insight into the mechanism of dynamic instability, based on high-resolution cryo-EM structures (4.7-5.6 Å) of dynamic microtubules and microtubules stabilized by GMPCPP or Taxol. We infer that hydrolysis leads to a compaction around the E-site nucleotide at longitudinal interfaces, as well as movement of the α-tubulin intermediate domain and H7 helix. Displacement of the C-terminal helices in both α- and β-tubulin subunits suggests an effect on interactions with binding partners that contact this region. Taxol inhibits most of these conformational changes, allosterically inducing a GMPCPP-like state. Lateral interactions are similar in all conditions we examined, suggesting that microtubule lattice stability is primarily modulated at longitudinal interfaces.
UR - http://www.scopus.com/inward/record.url?scp=84901362834&partnerID=8YFLogxK
U2 - 10.1016/j.cell.2014.03.053
DO - 10.1016/j.cell.2014.03.053
M3 - Article
C2 - 24855948
AN - SCOPUS:84901362834
SN - 0092-8674
VL - 157
SP - 1117
EP - 1129
JO - Cell
JF - Cell
IS - 5
ER -