SSBD - SSBD:database - Fig3A_PIP2_before

Detail of Fig3A_PIP2_before_EGF

SSBD:Repository

Title

The tracjectories of JF549–PI(4,5)P2 from colocalization of EGFR and PIP2 in living cells before EGF stimulation

Description

Time-lapse images of Halo-PI(4,5)P_{2} ( Halo-PLCdelta-PH) on HeLa cells before EGF (epidermal growth factor) stimulation. To simultaneously observe the dynamics of both EGFR(epidermal growth factor receptor) molecules and PI(4,5)P_{2} (phosphatidylinositol-4,5-bisphosphate) molecules upon EGF stimulation, EGFR-GFP and Halo-PI(4,5)P_{2} were imaged using dual-color total internal reflection (TIRF) microscopy. Halo-PI(4,5)P_{2} was visualized with JF549, a HaloTag ligand. This dataset includes only Halo-PI(4,5)P_{2} images. The images of Halo-PI(4,5)P_{2} detected by JF549 fluorescence were deposited in another dataset in this project; Fig3A_EGFR_before_EGF

Release, Updated

2026-06-09

Organism

Homo sapiens ( NCBI:txid9606 )

Strain(s)

Cell Line

HeLa cell

Molecular Function (MF)

phosphatidylinositol-4 , 5-bisphosphate binding

Biological Process (BP)

regulation of epidermal growth factor receptor signaling pathway

Cellular Component (CC)

plasma membrane

Biological Imaging Method

evanescent wave microscopy ( Fbbi:00000617 )

X scale

33.28 micrometer

Y scale

33.28 micrometer

T scale

30 milliseconds

Image Acquisition

Experiment type

Microscope type

Acquisition mode

Contrast method

Microscope model

Detector model

Objective model

Filter set

Summary of Methods

Abe M, Yanagawa M, Hiroshima M, Kobayashi T, Sako Y. Bilateral regulation of EGFR activity and local PI(4,5)P_{2} dynamics in mammalian cells observed with superresolution microscopy. Elife. 2024 Nov 8;13:e101652.

Related paper(s)

Mitsuhiro Abe, Masataka Yanagawa, Michio Hiroshima, Toshihide Kobayashi, Yasushi Sako (2024) Bilateral regulation of EGFR activity and local PI(4,5)P(2) dynamics in mammalian cells observed with superresolution microscopy., eLife, Volume 13

Published in 2024 Nov 8 (Electronic publication in Nov. 8, 2024, midnight )

doi: 10.7554/eLife.101652
pmid: 39513999

(Abstract) Anionic lipid molecules, including phosphatidylinositol-4,5-bisphosphate (PI(4,5)P(2)), are implicated in the regulation of epidermal growth factor receptor (EGFR). However, the role of the spatiotemporal dynamics of PI(4,5)P(2) in the regulation of EGFR activity in living cells is not fully understood, as it is difficult to visualize the local lipid domains around EGFR. Here, we visualized both EGFR and PI(4,5)P(2) nanodomains in the plasma membrane of HeLa cells using super-resolution single-molecule microscopy. The EGFR and PI(4,5)P(2) nanodomains aggregated before stimulation with epidermal growth factor (EGF) through transient visits of EGFR to the PI(4,5)P(2) nanodomains. The degree of coaggregation decreased after EGF stimulation and depended on phospholipase Cgamma, the EGFR effector hydrolyzing PI(4,5)P(2). Artificial reduction in the PI(4,5)P(2) content of the plasma membrane reduced both the dimerization and autophosphorylation of EGFR after stimulation with EGF. Inhibition of PI(4,5)P(2) hydrolysis after EGF stimulation decreased phosphorylation of EGFR-Thr654. Thus, EGFR kinase activity and the density of PI(4,5)P(2) around EGFR molecules were found to be mutually regulated.

(MeSH Terms)

Contact

Yasushi Sako , RIKEN , Cluster for Pioneering Research , Cluster for Pioneering Research