A fluorogenic complementation tool kit for interrogating lipid droplet-organelle interaction.

Autor:	Li X; Department of Cell and Molecular Biology, St. Jude Children's Research Hospital, Memphis, TN, USA., Gamuyao R; Department of Cell and Molecular Biology, St. Jude Children's Research Hospital, Memphis, TN, USA., Wu ML; Department of Cell and Molecular Biology, St. Jude Children's Research Hospital, Memphis, TN, USA., Cho WJ; Cell and Tissue Imaging Center, St. Jude Children's Research Hospital, Memphis, TN, USA., King SV; Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, TN, USA., Petersen RA; Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, TN, USA., Stabley DR; Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, TN, USA., Lindow C; Department of Cell and Molecular Biology, St. Jude Children's Research Hospital, Memphis, TN, USA., Climer LK; Department of Cell and Molecular Biology, St. Jude Children's Research Hospital, Memphis, TN, USA., Shirinifard A; Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, TN, USA., Ferrara F; Vector Production and Development Laboratory, St. Jude Children's Research Hospital, Memphis, TN, USA., Throm RE; Vector Production and Development Laboratory, St. Jude Children's Research Hospital, Memphis, TN, USA., Robinson CG; Cell and Tissue Imaging Center, St. Jude Children's Research Hospital, Memphis, TN, USA., Zhou Y; Department of Biostatistics, St. Jude Children's Research Hospital, Memphis, TN, USA., Carisey AF; Department of Cell and Molecular Biology, St. Jude Children's Research Hospital, Memphis, TN, USA., Tebo AG; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA., Chang CL; Department of Cell and Molecular Biology, St. Jude Children's Research Hospital, Memphis, TN, USA.
Jazyk:	angličtina
Zdroj:	The Journal of cell biology [J Cell Biol] 2024 Sep 02; Vol. 223 (9). Date of Electronic Publication: 2024 Jul 01.
DOI:	10.1083/jcb.202311126
Abstrakt:	Contact sites between lipid droplets and other organelles are essential for cellular lipid and energy homeostasis upon metabolic demands. Detection of these contact sites at the nanometer scale over time in living cells is challenging. We developed a tool kit for detecting contact sites based on fluorogen-activated bimolecular complementation at CONtact sites, FABCON, using a reversible, low-affinity split fluorescent protein, splitFAST. FABCON labels contact sites with minimal perturbation to organelle interaction. Via FABCON, we quantitatively demonstrated that endoplasmic reticulum (ER)- and mitochondria (mito)-lipid droplet contact sites are dynamic foci in distinct metabolic conditions, such as during lipid droplet biogenesis and consumption. An automated analysis pipeline further classified individual contact sites into distinct subgroups based on size, likely reflecting differential regulation and function. Moreover, FABCON is generalizable to visualize a repertoire of organelle contact sites including ER-mito. Altogether, FABCON reveals insights into the dynamic regulation of lipid droplet-organelle contact sites and generates new hypotheses for further mechanistical interrogation during metabolic regulation. (© 2024 Li et al.)
Databáze:	MEDLINE
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