title: Phospholipid Flippase ATP10A Translocates Phosphatidylcholine and Is Involved in Plasma Membrane Dynamics.
abstract: We showed previously that ATP11A and ATP11C have flippase activity toward aminophospholipids (phosphatidylserine (PS) and phosphatidylethanolamine (PE)) and ATP8B1 and that ATP8B2 have flippase activity toward phosphatidylcholine (PC) (Takatsu, H., Tanaka, G., Segawa, K., Suzuki, J., Nagata, S., Nakayama, K., and Shin, H. W. (2014) J. Biol. Chem. 289, 33543-33556). Here, we show that the localization of class 5 P4-ATPases to the plasma membrane (ATP10A and ATP10D) and late endosomes (ATP10B) requires an interaction with CDC50A. Moreover, exogenous expression of ATP10A, but not its ATPase-deficient mutant ATP10A(E203Q), dramatically increased PC flipping but not flipping of PS or PE. Depletion of CDC50A caused ATP10A to be retained at the endoplasmic reticulum instead of being delivered to the plasma membrane and abrogated the increased PC flipping activity observed by expression of ATP10A. These results demonstrate that ATP10A is delivered to the plasma membrane via its interaction with CDC50A and, specifically, flips PC at the plasma membrane. Importantly, expression of ATP10A, but not ATP10A(E203Q), dramatically altered the cell shape and decreased cell size. In addition, expression of ATP10A, but not ATP10A(E203Q), delayed cell adhesion and cell spreading onto the extracellular matrix. These results suggest that enhanced PC flipping activity due to exogenous ATP10A expression alters the lipid composition at the plasma membrane, which may in turn cause a delay in cell spreading and a change in cell morphology.
authors.affiliation: From the Graduate School and.
authors.forename: Tomoki
authors.initials: T
authors.lastname: Naito
authors.affiliation: From the Graduate School and.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Takatsu
authors.affiliation: From the Graduate School and.
authors.forename: Rie
authors.initials: R
authors.lastname: Miyano
authors.affiliation: the Faculty of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Naoto
authors.initials: N
authors.lastname: Takada
authors.affiliation: From the Graduate School and.
authors.forename: Kazuhisa
authors.initials: K
authors.lastname: Nakayama
authors.affiliation: From the Graduate School and shin@pharm.kyoto-u.ac.jp.
authors.forename: Hye-Won
authors.initials: HW
authors.lastname: Shin
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D002448:Cell Adhesion / Q000502:physiology; D002462:Cell Membrane / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D017931:DNA Primers; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism / Q000502:physiology; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D008969:Molecular Sequence Data; D010713:Phosphatidylcholines / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid; D013347:Subcellular Fractions / Q000201:enzymology
chemical_list: D017931:DNA Primers; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D010713:Phosphatidylcholines; C526978:TMEM30a protein, human; D000251:Adenosine Triphosphatases; C426648:ATP10A protein, human
keywords: ATPase; cell spreading; flippase; lipid bilayer; membrane protein; phospholipid; plasma membrane