2013 Fiscal Year Final Research Report
Development of super-resolution fluorescence microscopy and single-molecule analysis of signal transduction mechanism of a receptor protein in cell membrane
| Project Area | Integrative understanding of biological processes mediated by transient macromolecular complexes; New technology for visualizing physiologically metastable states. |
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| Project/Area Number |
21121004
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| Research Category |
Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
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| Allocation Type | Single-year Grants |
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| Review Section |
Complex systems
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| Research Institution | The University of Tokyo |
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Principal Investigator |
FUNATSU Takashi 東京大学, 薬学研究科(研究院), 教授 (00190124)
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| Co-Investigator(Renkei-kenkyūsha) |
KATO Takashi 早稲田大学, 教育・総合科学学術院, 教授 (80350388)
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| Project Period (FY) |
2009-07-23 – 2014-03-31
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| Keywords | １分子生理・生化学 / １分子イメージング / ナノ計測 / 受容体 / 生体膜 |
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| Research Abstract |
The thrombopoietin (TPO) receptor, Mpl, is a cytokine receptor regulating platelet production and early hematopoiesis. Although TPO signaling is initiated by phosphorylation of optimally oriented Mpl dimers, dimerization of Mpl is poorly understood. Here, we describe the regulatory mechanism of dimerization of Mpl by single-molecule fluorescence imaging in live cells. Mpl molecules were associated and dissociated repeatedly on the plasma membrane. As a result, Mpl existed as a mixture of monomers, dimers, and oligomers. TPO increased Mpl dimers or oligomers by stabilizing Mpl dimers. The stabilization of Mpl dimers was dependent on their phosphorylation and an adaptor protein, Shc, suggesting that phosphorylated Mpl dimers were crosslinked by Shc. In contrast, knockdown of Shc, an Mpl transmembrane mutant, influenced their phosphorylation. Thus, phosphorylation of Mpl was regulated by the stability of Mpl dimers in a positive feedback manner.
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[Journal Article] A spontaneously blinking fluorophore based on intramolecular spirocyclization for live-cell super-resolution imaging
Author(s)
Shin-nosuke Uno, Mako Kamiya, Toshitada Yoshihara, Ko Sugawara, Kohki Okabe, Mehmet C. Tarhan, Hiroyuki Fujita, Takashi Funatsu, Yashushi Okada, Seiji Tobita, Yasuteru Urano
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Journal Title
Nat. Chem
Volume: (In press)
Peer Reviewed
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