Lysosomal Interaction of Akt with Phafin2 : A Critical Step in the Induction of Autophagy

Matsuda-Lennikov Mami; Suizu Futoshi; Hirata Noriyuki; Hashimoto Manabu; Kimura Kohki; Nagamine Tadashi; Fujioka Yoichiro; Ohba Yusuke; Iwanaga Toshihiko; Noguchi Masayuki

Title	en Lysosomal Interaction of Akt with Phafin2 : A Critical Step in the Induction of Autophagy
Creator	en Matsuda-Lennikov, Mami en Suizu, Futoshi NRID 1000090431379 en Hirata, Noriyuki en Hashimoto, Manabu en Kimura, Kohki en Nagamine, Tadashi en Fujioka, Yoichiro en Ohba, Yusuke NRID 1000030333503 en Iwanaga, Toshihiko NRID 1000010160128 en Noguchi, Masayuki NRID 1000040359477
Accessrights	open access
Rights	http://creativecommons.org/licenses/by/3.0/ en Creative Commons Attribution 3.0 Unported
Subject	NDC 490
Description	Abstract en Autophagy is an evolutionarily conserved mechanism for the gross disposal of intracellular proteins in mammalian cells and dysfunction in this pathway has been associated with human disease. Although the serine threonine kinase Akt is suggested to play a role in this process, little is known about the molecular mechanisms by which Akt induces autophagy. Using a yeast two-hybrid screen, Phafin2 (EAPF or PLEKHF2), a lysosomal protein with a unique structure of N-terminal PH (pleckstrin homology) domain and C-terminal FYVE (Fab 1, YOTB, Vac 1, and EEA1) domain was found to interact with Akt. A sucrose gradient fractionation experiment revealed that both Akt and Phafin2 co-existed in the same lysosome enriched fraction after autophagy induction. Confocal microscopic analysis and BiFC analysis demonstrated that both Akt and Phafin2 accumulate in the lysosome after induction of autophagy. BiFC analysis using PtdIns (3) P interaction defective mutant of Phafin2 demonstrated that lysosomal accumulation of the Akt-Phafin2 complex and subsequent induction of autophagy were lysosomal PtdIns (3) P dependent events. Furthermore, in murine macrophages, both Akt and Phafin2 were required for digestion of fluorescent bacteria and/or LPS-induced autophagy. Taken together, these findings establish that lysosomal accumulation of Akt and Phafin2 is a critical step in the induction of autophagy via an interaction with PtdIns (3)P.
Publisher	en Public library science
Date	Issued2014-01-08
Language	eng
Resource Type	journal article
Version Type	VoR
Identifier	HDL http://hdl.handle.net/2115/54739
Relation	isIdenticalTo DOI https://doi.org/10.1371/journal.pone.0079795 PMID 24416124
Journal	PISSN 1932-6203 en Plos one Volume Number9 Issue Number1 Page Starte79795
File	fulltext PLoS One_9(1)_e79795.pdf 2.51 MB (application/pdf) Issued2014-01-08
Oaidate	2023-07-26