Journal
FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY
Volume 7, Issue -, Pages -Publisher
FRONTIERS MEDIA SA
DOI: 10.3389/fcell.2019.00181
Keywords
trans-Golgi network; cargo adaptor; sorting; clathrin; storm
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Funding
- Hong Kong Research Grants Council [26100315, 16101116, 16102218, AoE/M-05/123, C4002-17G, C4012-16E, HKUST12/CRF/13G]
- National Natural Science Foundation of China [NSFC 31871421]
- Shenzhen Science and Technology Innovation Committee [JCYJ20180306174847511]
- Offices of Provost
- VPRG
- Dean of Science, HKUST [HKUST VPRGO12SC02]
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The trans-Golgi network (TGN) and endosomes are essential protein sorting stations in the secretory transport pathway. Protein sorting is fundamentally a process of spatial segregation, but the spatial relationships among the proteins that constitute the sorting machinery have not been systematically analyzed at high resolution in mammalian cells. Here, using two-color STORM imaging, we show that the TGN/endosome-localized cargo adaptors, AP-1, GGA2 and epsinR, form elongated structures of over 250 nm in length at the juxta-nuclear Golgi area. Many of these structures are associated with clathrin. We found that AP-1 is spatially segregated from AP-3 and GGA2, whereas a fraction of AP-1 and GGA2 punctae are associated with epsinR. Moreover, we observed that the planar cell polarity cargo proteins, Vangl2 and Frizzled6 associate with different cargo adaptors-AP-1 and GGA2 or epsinR, respectively-when exiting the TGN. Knockdown analysis confirms the functional significance of this segregation. Our data indicates that TGN/endosome-localized cargo adaptors have distinct spatial relationships. The spatially segregated cargo adaptors GGA2 and AP-1 regulate sorting of Frizzled6 and Vangl2, respectively and spatially associated cargo adaptors can cooperatively regulate a specific sorting process.
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