Nucleation of Astral-shaped Microtubules from Latex Beads Conjugated with MTOG Proteins(Cell Biology)

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概要

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• A model system for the formation of astral-shaped microtubules (Mts) consisting of Latex beads (diameter of 0.2 μm), a protein fraction (p51) comprised of MTOGs (microtubule-organizing granules) and tubulin was established. The Latex beads were first incubated with p51 in the presence of GTP at 0℃, then the purified tubulin dimer fraction was added, resulting in the formation of an aster-like structure observed by dark-field microscopy. On preincubation of the Latex beads with GDP instead of GTP, the asters did not form. Unhydrolyzable GTP analogues such as GTP-γS and GMP-PNP promoted aster formation as did GTP as observed by dark-field microscopy. Polylysine, as representative of basic polymers capable of binding to the surface of the Latex beads, promoted spontaneous Mt assembly, and eventually an aster-like structure without Latex beads in the center formed. Further analyses made by measuring the optical density of the aster-forming system produced the following results. 1) preincubation of the Latex beads with GTP or GMP-PNP supported Mt assembly from the beads showing profiles typical for a sitedirected assembly without the lag phase. 2) GTP-γS and GDP inhibited the turbidity increase of the system, causing a decrease in both the initial velocity and the level of steady state of Mt assembly. 3) Anti-p51 monoclonal antibody (HP1) substantially inhibited the aster formation, while anti-γ-tubulin antibody only slightly inhibited assembly.

• 社団法人日本動物学会の論文
• 2000-07-01

著者

• OSUMI MASAKO

  Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University

• TAKAHASHI Kyoko

  Department of Food Science and Nutrition, School of Human Environmental Sciences, Mukogawa Women’s U

• Nakamura Akiko

  Department of Surgery, Kagoshima Medical Association Hospital

• OSUMI Masako

  日本女子大学理学部

• Osumi Masako

  Department Of Chemical And Biological Sciences Faculty Of Science Japan Women's University

• Osumi Masako

  Laboratory Of Electron Microscopy/open Research Center Japan Women's University

• Osumi Masako

  Department Of Chemical And Biological Sciences Faculty Of Science Japan Woman's University

• 大隈 正子

  日本女子大学理学部物質生物科学科

• 大隅 正子

  Department Of Chemical And Biological Sciences Faculty Of Science Japan Women's University

• Sato Mamiko

  Department Of Chemical And Biological Sciences Faculty Of Science Japan Women's University

• Sakai H

  Department Of Applied Aquabiology National Fisheries University

• SAKAI Hikoichi

  Department of Chemical and Biolobical Sciences, Faculty of Science, Japan Women's University

• Maruno Ayako

  Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University

• Sugawara Natsuki

  Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University

• Hoshi Chihiro

  Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University

• Nakamura Risako

  Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University

• Shinozaki Naeko

  Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University

• Maruno Ayako

  Department Of Chemical And Biological Sciences Faculty Of Science Japan Women's University

• Sugawara Natsuki

  Department Of Chemical And Biological Sciences Faculty Of Science Japan Women's University

• Nakamura Risako

  Department Of Chemical And Biological Sciences Faculty Of Science Japan Women's University

• Takahashi Kyoko

  Department Of Food Science And Nutrition School Of Human Environmental Sciences Mukogawa Women'

• Osumi Masako

  Department Of Biology Faculty Of Science Osaka City University

• Takahashi Kyoko

  Department Of Chemical And Biological Sciences Faculty Of Science Japan Women's University

• Hoshi Chihiro

  Department Of Chemical And Biological Sciences Faculty Of Science Japan Women's University

• Shinozaki Naeko

  Department Of Chemical And Biological Sciences Faculty Of Science Japan Women's University

• Nakamura Akiko

  Department Of Chemical And Biological Sciences Faculty Of Science Japan Women's University

• Nakamura Akiko

  Department Of Bioscience And Technology Faculty Of Agriculture Iwate University

• Sakai Hikoichi

  Department Of Biophysics And Biochemistry Faculty Of Science The University Of Tokyo

• Takahashi Kyoko

  Department Of Bioproduction Science Faculty Of Horticulture Chiba University

• NAKAMURA Akiko

  Department of Bacteriology, National Institute of Health

• SATO MAMIKO

  Department of Biology, Japan Women's University

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