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

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Venditti , R , Rega , L R , Masone , M C , Santoro , M , Polishchuk , E , Sarnataro , D , Paladino , S , D'Auria , S , Varriale , A , Olkkonen , V M , Di Tullio , G , Polishchuk , R & De Matteis , M A 2019 , ' Molecular determinants of ER-Golgi contacts identified through a new FRET-FLIM system ' , Journal of Cell Biology , vol. 218 , no. 3 , pp. 1055-1065 . https://doi.org/10.1083/jcb.201812020

Title: Molecular determinants of ER-Golgi contacts identified through a new FRET-FLIM system
Author: Venditti, Rossella; Rega, Laura Rita; Masone, Maria Chiara; Santoro, Michele; Polishchuk, Elena; Sarnataro, Daniela; Paladino, Simona; D'Auria, Sabato; Varriale, Antonio; Olkkonen, Vesa M.; Di Tullio, Giuseppe; Polishchuk, Roman; De Matteis, Maria Antonietta
Other contributor: University of Helsinki, Medicum



Date: 2019-03
Language: eng
Number of pages: 11
Belongs to series: Journal of Cell Biology
ISSN: 0021-9525
DOI: https://doi.org/10.1083/jcb.201812020
URI: http://hdl.handle.net/10138/305645
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.
Subject: ENDOPLASMIC-RETICULUM
PLASMA-MEMBRANE
LIPID-TRANSFER
BINDING
PHOSPHATIDYLSERINE
VAP
TRAFFICKING
TRANSPORT
PROTEINS
INTERACTOME
3111 Biomedicine
1182 Biochemistry, cell and molecular biology
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