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Production of PtdInsP3 at endomembranes is triggered by receptor endocytosis

Nature Cell Biology volume 5, pages 1016–1022 (2003)Cite this article

Abstract

Phosphatidylinositol-3,4,5-trisphosphate (PtdInsP3) regulates diverse cellular functions, including cell proliferation and apoptosis, and has roles in the progression of diabetes and cancer. However, little is known about its production. Here, we describe fluorescent indicators for PtdInsP3 that allow a spatio-temporal examination of PtdInsP3 production in single living cells. After ligand stimulation, PtdInsP3 levels increased to a larger extent at the endomembranes (that is, the endoplasmic reticulum and the Golgi) than at the plasma membrane. This increase was found to originate from in situ production at the endomembranes, a process stimulated directly by receptor tyrosine kinases endocytosed from the plasma membrane to the endomembranes. The demonstration of PtdInsP3 production through receptor endocytosis addresses a long-standing question about how signalling pathways downstream of PtdInsP3 are activated at intracellular compartments remote from the plasma membrane.

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Figure 1: Fluorescent indicators for PtdInsP3 in single living cells.
Figure 2: Fluorescence imaging of fllip-pm.
Figure 3: Response of fllip-em to PtdInsP3 at the endomembranes after PDGF stimulation.
Figure 4: Inhibition of PtdInsP3 production at endomembranes.

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Acknowledgements

This work was supported by CREST (Core Research for Evolutional Science and Technology) of JST (Japan Science and Technology Agency) and grants for M.S. and Y.U. from the Ministry of Education, Science and Culture, Japan. We thank J. E. Pessin for providing an adenovirus vector of DynK44A, M. Bryer-Ash for providing an adenovirus vector of PTP1B and Y. Imai for experimental help.

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  1. Moritoshi Sato and Yoshibumi Ueda: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry, School of Science, The University of Tokyo, and Japan Science and Technology Agency, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan

    Moritoshi Sato, Yoshibumi Ueda, Tokio Takagi & Yoshio Umezawa

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  1. Moritoshi Sato
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Correspondence to Yoshio Umezawa.

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Sato, M., Ueda, Y., Takagi, T. et al. Production of PtdInsP3 at endomembranes is triggered by receptor endocytosis. Nat Cell Biol 5, 1016–1022 (2003). https://doi.org/10.1038/ncb1054

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