Microtubule structure at improved resolution.
Microtubule architecture can vary with eukaryotic species, with different cell types, and with the presence of stabilizing agents. For in vitro assembled microtubules, the average number of protofilaments is reduced by the presence of sarcodictyin A, epothilone B, and eleutherobin (similarly to taxol) but increased by taxotere. Assembly with a slowly hydrolyzable GTP analogue GMPCPP is known to give 96% 14 protofilament microtubules. We have used electron cryomicroscopy and helical reconstruction techniques to obtain three-dimensional maps of taxotere and GMPCPP microtubules incorporating data to 14 A resolution. The dimer packing within the microtubule wall is examined by docking the tubulin crystal structure into these improved microtubule maps. The docked tubulin and simulated images calculated from "atomic resolution" microtubule models show tubulin heterodimers are aligned head to tail along the protofilaments with the beta subunit capping the microtubule plus end. The relative positions of tubulin dimers in neighboring protofilaments are the same for both types of microtubule, confirming that conserved lateral interactions between tubulin subunits are responsible for the surface lattice accommodation observed for different microtubule architectures. Microtubules with unconventional protofilament numbers that exist in vivo are likely to have the same surface lattice organizations found in vitro. A curved "GDP" tubulin conformation induced by stathmin-like proteins appears to weaken lateral contacts between tubulin subunits and could block microtubule assembly or favor disassembly. We conclude that lateral contacts between tubulin subunits in neighboring protofilaments have a decisive role for microtubule stability, rigidity, and architecture.
Meurer-Grob
P
P
Institut de Biologie Structurale (CEA/CNRS), 41 rue Jules Horowitz, 38027 Grenoble, France.
Kasparian
J
J
Wade
R H
RH
D008841:Actin Cytoskeleton / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D000470:Alkaloids / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D019281:Dimerization; D004224:Diterpenes; D000077143:Docetaxel; D034261:Epothilones; D004852:Epoxy Compounds / Q000378:metabolism; D005079:Excipients / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D008870:Microtubules / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D008969:Molecular Sequence Data; D017239:Paclitaxel / Q000031:analogs & derivatives / Q000378:metabolism; D011487:Protein Conformation; D013499:Surface Properties; D013552:Swine; D043823:Taxoids; D013844:Thiazoles / Q000378:metabolism; D014404:Tubulin / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure
D000470:Alkaloids; D004224:Diterpenes; D034261:Epothilones; D004852:Epoxy Compounds; D005079:Excipients; D043823:Taxoids; D013844:Thiazoles; D014404:Tubulin; C109551:eleutherobin; C109552:sarcodictyin A; D000077143:Docetaxel; C004805:5'-guanylylmethylenebisphosphonate; D006160:Guanosine Triphosphate; D017239:Paclitaxel; C093788:epothilone B