title: X-ray fiber diffraction analysis shows dynamic changes in axial tubulin repeats in native microtubules depending on paclitaxel content, temperature and GTP-hydrolysis.
abstract: Microtubules are key components of the cytoskeleton in eukaryotic cells. The dynamics between assembled microtubules and free tubulin dimers in the cytoplasm is closely related to the active shape changes of microtubule networks. One of the most fundamental questions is the association of microtubule dynamics with the molecular conformation of tubulin within microtubules. To address this issue, we applied a new technique for the rapid shear-flow alignment of biological filaments, enabling us to acquire the structural periodicity data of microtubules by X-ray fiber diffraction under various physiological conditions. We classified microtubules into three main groups on the basis of distinct axial tubulin periodicities and mean microtubule diameters that varied depending on GTP hydrolysis and the content of paclitaxel, a microtubule stabilizer. Paclitaxel induced rapid changes in tubulin axial repeats in a cooperative manner. This is the first demonstration of dynamic changes of axial tubulin repeats within native microtubules without fixation. We also found extraordinary features of negative thermal expansion of axial tubulin repeats in both paclitaxel-stabilized and GMPCPP-containing microtubules. Our results suggest that even in assembled microtubules, both GTP- and GDP-tubulin dimers can undergo dynamic conversion between at least two different states: short and long configurations.
authors.affiliation: Department of Biological Sciences, Faculty of Science and Engineering, Chuo University, Kasuga, Bunkyo, Tokyo, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Kamimura
authors.affiliation: Department of Biological Sciences, Faculty of Science and Engineering, Chuo University, Kasuga, Bunkyo, Tokyo, Japan.
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Fujita
authors.affiliation: Department of Biological Sciences, Faculty of Science and Engineering, Chuo University, Kasuga, Bunkyo, Tokyo, Japan.
authors.forename: Yuuko
authors.initials: Y
authors.lastname: Wada
authors.affiliation: Department of Life Sciences, Faculty of Life and Environmental Sciences, Prefectural University of Hiroshima, Shobara, Hiroshima, Japan.
authors.forename: Toshiki
authors.initials: T
authors.lastname: Yagi
authors.affiliation: Life and Environmental Division, SPring-8, Japan Synchrotron Radiation Research Institute (JASRI), Sayo, Hyogo, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Iwamoto
mesh_terms: D000818:Animals; D006160:Guanosine Triphosphate / Q000737:chemistry*; D006868:Hydrolysis; D008870:Microtubules / Q000737:chemistry* / Q000648:ultrastructure*; D017239:Paclitaxel / Q000737:chemistry*; D013552:Swine; D014404:Tubulin / Q000737:chemistry* / Q000378:metabolism; D014961:X-Ray Diffraction
chemical_list: D014404:Tubulin; D006160:Guanosine Triphosphate; D017239:Paclitaxel
keywords: GMPCPP; X-ray fiber diffraction analysis; microtubule structure in solution; negative thermal expansion of microtubules; paclitaxel; shear-flow alignment of microtubules