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bioRxiv · 10.1101/2023.08.14.553257

ATG2A-mediated bridge-like lipid transport regulates lipid droplet accumulation

Korfhage, J. L.·Wan, N.·Elhan, H.·Kauffman, L.·Pineda, M. J.·Fuller, D. M.·Thiam, A. R.·Reinisch, K.·Melia, T.
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Abstract

ATG2 proteins facilitate bulk lipid transport between membranes. ATG2 is an essential autophagy protein, but ATG2 also localizes to lipid droplets (LDs), and genetic depletion of ATG2 increases LD numbers while impairing fatty acid transport from LDs to mitochondria. How ATG2 supports LD homeostasis and whether lipid transport regulates this homeostasis remains unknown. Here we demonstrate that ATG2 is preferentially recruited to phospholipid monolayers such as those surrounding LDs rather than to phospholipid bilayers. In vitro, ATG2 can drive phospholipid transport from artificial LDs with rates that correlate with the binding affinities, such that phospholipids are moved much more efficiently when one of the ATG2-interacting structures is an artificial LD. ATG2 is thought to exhibit bridge-like" lipid transport, with lipids flowing across the protein between membranes. We mutated key amino acids within the bridge to form a transport-dead ATG2 mutant (TD-ATG2A) which we show specifically blocks bridge-like, but not shuttle-like, lipid transport in vitro. TD-ATG2A still localizes to LDs, but is unable to rescue LD accumulation in ATG2 knockout cells. Thus, ATG2 has a natural affinity for, and an enhanced activity upon LD surfaces and uses bridge-like lipid transport to support LD dynamics in cells.

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Korfhage, J. L., Wan, N., Elhan, H., Kauffman, L., Pineda, M. J., Fuller, D. M., Thiam, A. R., Reinisch, K., Melia, T.. 2023-08-14. ATG2A-mediated bridge-like lipid transport regulates lipid droplet accumulation. https://doi.org/10.1101/2023.08.14.553257

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