TY - JOUR
T1 - Single-molecule imaging of PI(4,5)P2 and PTEN in vitro reveals a positive feedback mechanism for PTEN membrane binding
AU - Yoshioka, Daisuke
AU - Fukushima, Seiya
AU - Koteishi, Hiroyasu
AU - Okuno, Daichi
AU - Ide, Toru
AU - Matsuoka, Satomi
AU - Ueda, Masahiro
N1 - Funding Information:
GFP-Nodulin was kindly provided by Y. Miao and P. N. Devreotes (Johns Hopkins University School of Medicine, Baltimore). We thank T. Uyeda (National Institute of Advanced Industrial Science and Technology, Ibaraki, Japan) for the PHAKT/PKB-EGFP construct, and members of the Ueda laboratories for helpful suggestions. We thank P. Karagiannis for English editing (Sofia Science Writing, Japan). This research was supported by Japan Society for the Promotion of Science (JSPS) KAKENHI Grant JP25871120 (to S.M.), by Precursory Research for Embryonic Science and Technology (JST-PRESTO) from Japan Science and Technology Agency JPMJPR1879 (to S.M.), and by the Advanced Research and Development Programs for Medical Innovation (AMED-CREST) from Japan Agency for Medical Research and Development AMED JP18gm0910001 (to M.U.).
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - PTEN, a 3-phosphatase of phosphoinositide, regulates asymmetric PI(3,4,5)P3 signaling for the anterior-posterior polarization and migration of motile cells. PTEN acts through posterior localization on the plasma membrane, but the mechanism for this accumulation is poorly understood. Here we developed an in vitro single-molecule imaging assay with various lipid compositions and use it to demonstrate that the enzymatic product, PI(4,5)P2, stabilizes PTEN’s membrane-binding. The dissociation kinetics and lateral mobility of PTEN depended on the PI(4,5)P2 density on artificial lipid bilayers. The basic residues of PTEN were responsible for electrostatic interactions with anionic PI(4,5)P2 and thus the PI(4,5)P2-dependent stabilization. Single-molecule imaging in living Dictyostelium cells revealed that these interactions were indispensable for the stabilization in vivo, which enabled efficient cell migration by accumulating PTEN posteriorly to restrict PI(3,4,5)P3 distribution to the anterior. These results suggest that PI(4,5)P2-mediated positive feedback and PTEN-induced PI(4,5)P2 clustering may be important for anterior-posterior polarization.
AB - PTEN, a 3-phosphatase of phosphoinositide, regulates asymmetric PI(3,4,5)P3 signaling for the anterior-posterior polarization and migration of motile cells. PTEN acts through posterior localization on the plasma membrane, but the mechanism for this accumulation is poorly understood. Here we developed an in vitro single-molecule imaging assay with various lipid compositions and use it to demonstrate that the enzymatic product, PI(4,5)P2, stabilizes PTEN’s membrane-binding. The dissociation kinetics and lateral mobility of PTEN depended on the PI(4,5)P2 density on artificial lipid bilayers. The basic residues of PTEN were responsible for electrostatic interactions with anionic PI(4,5)P2 and thus the PI(4,5)P2-dependent stabilization. Single-molecule imaging in living Dictyostelium cells revealed that these interactions were indispensable for the stabilization in vivo, which enabled efficient cell migration by accumulating PTEN posteriorly to restrict PI(3,4,5)P3 distribution to the anterior. These results suggest that PI(4,5)P2-mediated positive feedback and PTEN-induced PI(4,5)P2 clustering may be important for anterior-posterior polarization.
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U2 - 10.1038/s42003-020-0818-3
DO - 10.1038/s42003-020-0818-3
M3 - Article
C2 - 32111929
AN - SCOPUS:85080985874
SN - 2399-3642
VL - 3
JO - Communications Biology
JF - Communications Biology
IS - 1
M1 - 92
ER -