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Differential requirement for ATG2A domains for localization to autophagic membranes and lipid droplets.

FEBS letters (2017-11-08)
Norito Tamura, Taki Nishimura, Yuriko Sakamaki, Ikuko Koyama-Honda, Hayashi Yamamoto, Noboru Mizushima
ABSTRACT

ATG2 is one of the autophagy-related (ATG) proteins essential for autophagosome formation and localizes to isolation membranes and lipid droplets in mammalian cells. Here, we investigated the requirement of regions in ATG2A for its organellar localization and function. The N-terminal amino acids 1-198 and the C-terminal amino acids 1830-1938 are required for the localization to isolation membranes and lipid droplets, respectively. The C-terminal region is not required for the localization to isolation membranes and for autophagy. We also identified an amphipathic helix in ATG2A that is required for both its localization to organelles and autophagosome formation. These data suggest that the dual localization of ATG2A is regulated by different regions.

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Terreno Eagle modificato di Dulbecco - alto glucosio, With 4500 mg/L glucose, sodium pyruvate, and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
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Hexadimethrine bromide, ≥94% (titration)
Sigma-Aldrich
Anti-Atg18 (WIPI-2) Antibody, clone 2A2, clone 2A2, from mouse
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Anti-WIPI-2 (C-terminal) antibody produced in rabbit, ~1.0 mg/mL, affinity isolated antibody