| Project/Area Number |
03454009
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
植物生理学
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| Research Institution | Osaka University |
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Principal Investigator |
SHIBAOKA Hiroh Faculty of Science, Osaka University, Professor, 理学部, 教授 (60087054)
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| Co-Investigator(Kenkyū-buntansha) |
SONOBE Seiji Faculty of Science, Osaka University, Assistant, 理学部, 助手 (30197024)
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| Project Period (FY) |
1991 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥5,700,000 (Direct Cost: ¥5,700,000)
Fiscal Year 1992: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1991: ¥3,000,000 (Direct Cost: ¥3,000,000)
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| Keywords | Gibberellin / Microtubules / Cultured cell / Cross-bridge protein / Plant tubulin / Cell biology / 膜ゴースト / 微小管・細胞膜架橋構造 / プロトプラスト / 微小管の安定性 |
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| Research Abstract |
1. Isolation of a microtubule-plasma membrane cross-linking protein from tobacco cultured cells - Tubulin in the extract prepared from miniprotoplasts that had been prepared from protoplasts of tobacco BY-2 cells polymerized into microtubules in the absence of any microtubule-stabilizing agents. The polymerized microtubules formed bundles and between adjacent microtubules in the bundle cross-bridges were observed. A protein of 65Kd was copurified with tubulin by assembly and disassembly method. The protein was found to have the ability to cross-link microtubules. Immunofluorescence microscopy using antibodies against the 65Kd protein revealed that this protein is codistributed with microtubules in tobacco BY-2 cells.
2. Cross-bridge structures between microtubules and plasma membrane in tobacco cultured cells - Membrane ghosts prepared from tobacco BY-2 cells have the ability to bind microtubules that were polymerized in the extract prepared from tobacco BY-2 miniprotoplasts. Treatment with 0.6 M KCl or trypsin eliminated the ability of the ghosts to bind microtubules, indicating a factor that is necessary for the binding is protein and is extracted by high concentrations of KCl. In the presence of taxol, a microtubule-stabilizing agent, microtubules polymerized in the extract were bound to the ghosts. The factor eliminated by KCl seems to have a microtubule-stabilizing activity. Microtubules polymerized in the extract attached to the ghost in the presence of taxol, but microtubules polymerized from purified tubulin didn't, indicating the presence in the extract of factor(s) that is necessary for the attachment of microtubules to the membrane.
3. Microtubule-stabilizing effect of gibberellin A_3 (GA_3) in maize mesocotyl cells - GA_3 stabilized microtubules in mesocotyl cells of maize seedlings. Microtubules in protoplasts prepared from mesocotyls of GA_3-treated seedlings were more stable than those in protoplasts prepared from GA_3-untreated seedlings.
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