ESCRT-0 marks an APPL1-independent transit route for EGFR between the cell surface and the EEA1-positive early endosome
| Autor: | Neftali, Flores-Rodriguez, David A, Kenwright, Pei-Hua, Chung, Andrew W, Harrison, Flavia, Stefani, Thomas A, Waigh, Victoria J, Allan, Philip G, Woodman |
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| Rok vydání: | 2015 |
| Předmět: |
Hrs
Particle-based colocalisation EGFR Vesicular Transport Proteins macromolecular substances Endosomes EEA1 Cell Line Tumor Image Processing Computer-Assisted Humans RNA Small Interfering Transport Vesicles Sorting Nexins Adaptor Proteins Signal Transducing Endosomal Sorting Complexes Required for Transport Epidermal Growth Factor Cell Membrane Ubiquitination Phosphoproteins Enzyme Activation ErbB Receptors Protein Transport ESCRT-I RNA Interference HeLa Cells Research Article |
| Zdroj: | Journal of Cell Science |
| ISSN: | 1477-9137 |
| Popis: | Endosomal sorting complexes required for transport (ESCRT)-0 sorts ubiquitylated EGFR within the early endosome so that the receptor can be incorporated into intralumenal vesicles. An important question is whether ESCRT-0 acts solely upon EGFR that has already entered the vacuolar early endosome (characterised by the presence of EEA1) or engages EGFR within earlier compartments. Here, we employ a suite of software to determine the localisation of ESCRT-0 at subpixel resolution and to perform particle-based colocalisation analysis with other endocytic markers. We demonstrate that although some of the ESCRT-0 subunit Hrs (also known as HGS) colocalises with the vacuolar early endosome marker EEA1, most localises to a population of peripheral EEA1-negative endosomes that act as intermediates in transporting EGFR from the cell surface to more central early endosomes. The peripheral Hrs-labelled endosomes are distinct from APPL1-containing endosomes, but co-label with the novel endocytic adaptor SNX15. In contrast to ESCRT-0, ESCRT-I is recruited to EGF-containing endosomes at later times as they move to more a central position, whereas ESCRT-III is also recruited more gradually. RNA silencing experiments show that both ESCRT-0 and ESCRT-I are important for the transit of EGF to EEA1 endosomes. |
| Databáze: | OpenAIRE |
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