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Phospholipid Flippases Lem3p-Dnf1p and Lem3p-Dnf2p Are Involved in the Sorting of the Tryptophan Permease Tat2p in Yeast
| Title: | Phospholipid Flippases Lem3p-Dnf1p and Lem3p-Dnf2p Are Involved in the Sorting of the Tryptophan Permease Tat2p in Yeast |
| Authors: | Hachiro, Takeru Browse this author | | Yamamoto, Takaharu Browse this author →KAKEN DB | | Nakano, Kenji Browse this author | | Tanaka, Kazuma Browse this author →KAKEN DB |
| Keywords: | Amino Acid Transport | | Membrane Bilayer | | Phosphatidylserine | | Protein Sorting | | Ubiquitination | | Yeast | | Flippase | | Phospholipid Asymmetry | | Type 4 P-type ATPase |
| Issue Date: | 1-Feb-2013 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 288 |
| Issue: | 5 |
| Start Page: | 3594 |
| End Page: | 3608 |
| Publisher DOI: | 10.1074/jbc.M112.416263 |
| Abstract: | The type 4 P-type ATPases are flippases that generate phospholipid asymmetry in membranes. In budding yeast, heteromeric flippases, including Lem3p-Dnf1p and -Dnf2p translocate phospholipids to the cytoplasmic leaflet of membranes. Here we report that Lem3p-Dnf1/2p are involved in transport of the tryptophan permease Tat2p to the plasma membrane. The lem3Δ mutant exhibited a tryptophan requirement due to the mislocalization of Tat2p to intracellular membranes. Tat2p was relocalized to the plasma membrane when trans-Golgi network (TGN)-to-endosome transport was inhibited. Inhibition of ubiquitination by mutations in ubiquitination machinery also rerouted Tat2p to the plasma membrane. Lem3p-Dnf1/2p are localized to endosomal/TGN membranes in addition to the plasma membrane. Endocytosis mutants, in which Lem3p-Dnf1/2p are sequestered to the plasma membrane. Endocytosis mutants, in which Lem3p-Dnf1/2p are sequestered to the plasma membrane, also exhibited the ubiquitination-dependent missorting of Tat2p. These results suggest that Tat2p is ubiquitinated at the TGN and missorted to the vacuolar pathway in the lem3Δ mutant. The NH2-terminal cytoplasmic region of Tat2p containing ubiquitination acceptor lysines interacted with liposomes containing acidic phospholipids, including phosphatidylserine. This interaction was abrogated by alanine substitution mutations in the basic amino acids downstream of the ubiquitination sites. Interestingly, a mutant Tat2p containing these substitutions was missorted in a ubiquitination-dependent manner. We propose the following model based on these results; Tat2p is not ubiquitinated when the NH2-terminal region is bound to membrane phospholipids, but if it dissociates from the membrane due to a low level of phosphatidylserine caused by perturbation of phospholipid asymmetry in the lem3Δ mutant, Tat2p is ubiquitinated and then transported from the TGN to the vacuole. |
| Rights: | This research was originally published in Journal of Biological Chemistry. Takeru Hachiro; Takaharu Yamamoto; Kenji Nakano; Kazuma Tanaka. Phospholipid Flippases Lem3p-Dnf1p and Lem3p-Dnf2p Are Involved in the Sorting of the Tryptophan Permease Tat2p in Yeast. Journal of Biological Chemistry. 2013; 288:3594-3608. © the American Society for Biochemistry and Molecular Biology. |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/52118 |
| Appears in Collections: | 遺伝子病制御研究所 (Institute for Genetic Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 鉢呂 健
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