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Phosphatidylinositol 3-phosphate is found in microdomains of early endosomes

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Abstract

Phosphatidylinositol 3-phosphate [PI(3)P] is a phosphatidylinositol 3-kinase product whose localisation is restricted to the limiting membranes of early endosomes and to the internal vesicles of multivesicular bodies. In this study the intracellular distribution of PI(3)P was compared with those of another phosphoinositide and a number of endosomal proteins. Using a 2xFYVE probe specific for PI(3)P we found that PI(3)P is present in microdomains within the endosome membrane, whereas a phosphoinositide required for clathrin-mediated endocytosis, PI(4,5)P2, was only detected at the plasma membrane. The small GTPase Rab5 as well as the PI(3)P-binding proteins EEA1, SARA and CISK were found to be abundant within PI(3)P-containing endosomal microdomains. In contrast, another PI(3)P-binding protein, Hrs, was found concentrated in clathrin-coated endosomal microdomains with low levels of PI(3)P. While PI(3)P-containing microdomains could be readily distinguished on enlarged endosomes in cells transfected with a constitutively active Rab5 mutant, such domains could also be detected in endosomes of non-transfected cells. We conclude that the membranes of early endosomes consist of microdomains in which PI(3)P and specific proteins are concentrated. These microdomains may be necessary for the assembly of distinct multimolecular complexes that specify organelle identity, membrane trafficking and receptor signalling.

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Acknowledgements

We thank Eva Rønning for technical assistance. This study was supported by the Research Council of Norway, the Norwegian Cancer Society and the Novo Nordisk Foundation.

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Authors and Affiliations

  1. Department of Biochemistry, Institute for Cancer Research, The Norwegian Radium Hospital, Montebello, 0310, Oslo, Norway

    David J. Gillooly, Camilla Raiborg & Harald Stenmark

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  1. David J. Gillooly
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  2. Camilla Raiborg
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  3. Harald Stenmark
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Correspondence to Harald Stenmark.

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David J. Gillooly and Camilla Raiborg contributed equally

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Gillooly, D.J., Raiborg, C. & Stenmark, H. Phosphatidylinositol 3-phosphate is found in microdomains of early endosomes. Histochem Cell Biol 120, 445–453 (2003). https://doi.org/10.1007/s00418-003-0591-7

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