2005 Fiscal Year Final Research Report Summary
Molecular mechanism of AAA-type ATPase dynein
| Project/Area Number |
16370069
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biophysics
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| Research Institution | The University of Tokyo |
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Principal Investigator |
TOYOSHIMA Yoko Y. The University of Tokyo, Department of Life Sciences, Associate Professor, 大学院・総合文化研究科, 助教授 (40158043)
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| Co-Investigator(Kenkyū-buntansha) |
EDAMATSU Masaki The University of Tokyo, Department of Life Sciences, Research Associate, 大学院・総合文化研究科, 助手 (60251328)
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| Project Period (FY) |
2004 – 2005
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| Keywords | Dynein / Molecular motor / Microtubules / Stalk / AAA motif / ATPase activity / ATP binding site / Microtubule binding site |
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| Research Abstract |
Motility of cytoplasmic dynein has been studied using in vitro microtubule-gliding assay system. Single molecules of dynein stalk head, as well as the stalk fragment composed of stalk head and the coiled-coil region, were observed to attach to the microtubule, move on the microtubule, and then detach from the microtubule. Analysis of the movement revealed to be Brownian motion, suggesting that the binding force of the stalk fragments is weak. Native dynein molecules purified from porcine brain were visualized by binding of the recombinant p150 fragment which contained dynein binding region and fused with gelsolin. In the presence of ATP, single molecules of native dynein did not show processive movement long enough to detect with a regular speed video camera (30fps). However, they behaved differently depending on the nucleotide conditions used. The native dynein molecules sit firmly on microtubules in the presence of AMP-PNP or ADP, and also in the absence of nucleotides. Meanwhile, in the presence of ADP-Vi, they moved around along microtubules very similar way to the movement of the stalk fragments. These results suggest that single molecules of dynein weakly bind to and keep contact with microtubules in ADP-Pi state, and the isolated stalk fragments mimic this state. The microtubule binding fragments of dynactin p150 also showed Brownian motion along microtubules, and the lifetime on microtubules were much longer than those of the stalk fragments and the native dynein molecules. These results show that dynactin p150 can contribute to increase the processivity of dynein movement without disturbing the motility in dynein-dynactin complex.
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[Journal Article] A single-headed recombinant fragment of Dictyostelium cyoplasmic dynein can drive the robust sliding of microtubules2004
Author(s)
Nishiura, N., Kon, T., Shiroguchi, K., Ohkura, R., Shima, T., Toyoshima, Y.Y., Sutoh, K.
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Journal Title
J. Biol. Chem. 279
Pages: 22799-22802
Description
「研究成果報告書概要(和文)」より
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[Journal Article] Dynein and kinesin share an overlapping microtubule-binding site2004
Author(s)
Miuno, N., Toba, S., Edamatsu, M., Watai-Nishii, J., Hirokawa, N., Toyoshima, Y.Y., Kikkawa, M.
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Journal Title
EMBO J. 23
Pages: 2459-2467
Description
「研究成果報告書概要(和文)」より
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[Journal Article] A single-headed recombinant fragment of Dictyostelium cytoplasmic dynein can drive the robust sliding of microtubules2004
Author(s)
Nishiura, N., Kon, T., Shiroguchi, K., Ohkura, R., Shima, T., Toyoshima, Y.Y., Sutoh, K.
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Journal Title
J.Biol. Chem. 279
Pages: 22799-22802
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Dynein and kinesin share an overlapping microtubule-binding site2004
Author(s)
Mizuno, N., Toba, S., Edamatsu, M., Watai-Nishii, J., Hirokawa, N., Toyoshima, Y.Y., Kikkawa, M.
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Journal Title
EMBO J. 23
Pages: 2459-2467
Description
「研究成果報告書概要(欧文)」より
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