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J Cell Biol. 2019 Mar 4;218(3):1055-1065. doi: 10.1083/jcb.201812020. Epub 2019 Jan 18.

Molecular determinants of ER-Golgi contacts identified through a new FRET-FLIM system.

Author information

1
Telethon Institute of Genetics and Medicine, Pozzuoli, Italy venditti@tigem.it.
2
Department of Molecular Medicine and Medical Biotechnology, University of Napoli Federico II, Medical School, Naples, Italy.
3
Telethon Institute of Genetics and Medicine, Pozzuoli, Italy.
4
Institute of Food Science, Consiglio Nazionale delle Ricerche, Avellino, Italy.
5
Department of Anatomy, Faculty of Medicine, FI-00014 University of Helsinki, Helsinki, Finland.
6
Minerva Foundation Institute for Medical Research, Biomedicum 2U Helsinki, Helsinki, Finland.
7
Telethon Institute of Genetics and Medicine, Pozzuoli, Italy dematteis@tigem.it.

Abstract

ER-TGN contact sites (ERTGoCS) have been visualized by electron microscopy, but their location in the crowded perinuclear area has hampered their analysis via optical microscopy as well as their mechanistic study. To overcome these limits we developed a FRET-based approach and screened several candidates to search for molecular determinants of the ERTGoCS. These included the ER membrane proteins VAPA and VAPB and lipid transfer proteins possessing dual (ER and TGN) targeting motifs that have been hypothesized to contribute to the maintenance of ERTGoCS, such as the ceramide transfer protein CERT and several members of the oxysterol binding proteins. We found that VAP proteins, OSBP1, ORP9, and ORP10 are required, with OSBP1 playing a redundant role with ORP9, which does not involve its lipid transfer activity, and ORP10 being required due to its ability to transfer phosphatidylserine to the TGN. Our results indicate that both structural tethers and a proper lipid composition are needed for ERTGoCS integrity.

PMID:
30659100
PMCID:
PMC6400564
[Available on 2019-09-04]
DOI:
10.1083/jcb.201812020

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