459-Abe-EGFRdynamics
Author reviewedTIRF imaging of EGFR and PIP2 on Hela cells
Project ID
459-Abe-EGFRdynamics
Project SID
459
Title
TIRF imaging of EGFR and PIP2 on Hela cells
Description
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.
Submission Date
2025-07-30
Opened Date
-
Release Date
2026-06-09
Update Date
-
Kind
Image datasets and Quantitative datasets
License
Project URL
-
Project DOI
-
Metadata Version
-
Template Version
3.0.2
Funding Information
This research was supported by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (22K06609 to MA, 19H05647 to YS); Japan Science and Technology Agency (JST), PRESTO, JPMJPR20EF (MY); and the Glycolipidologue Program of RIKEN (to YS and TK).
Total
8 files / 636.4 MB
Image datasets
zipped 4 files / 236.3 MB, raw image 4 files / 400.1 MB, total 8 files / 636.4 MB
Quantitative datasets
zipped 0 files / 0 bytes, raw bdml 0 files / 0 bytes, total 0 files / 0 bytes
Biosample
Organism
Homo sapiens
(
NCBITaxon:9606
)
Strain
-
Cell
-
Cell line
HeLa cell
(
CLO:0003684
)
MeSH
Ontology
Anatomical Entity
-
UBERON
-
Biological Process
response to epidermal growth factor
(
GO:0070849
)
regulation of epidermal growth factor receptor signaling pathway
(
GO:0042058
)
Cellular Component
plasma membrane
(
GO:0005886
)
Molecular Function
epidermal growth factor receptor activity
(
GO:0005006
)
phosphatidylinositol-4,5-bisphosphate binding
(
GO:0005546
)
Medical Subject Headings
Imaging Method
Method involved in biological imaging
evanescent wave microscopy
(
FBbi:00000617
)
Paper DOI / Paper URL
People
Contact
Yasushi Sako
RIKEN
Cluster for Pioneering Research
Cellular Informatics Laboratory
2-1, Hirosawa, Wako, Saitama 351-0198, Japan
Imaging dataset contributor
Mitsuhiro Abe (RIKEN)
Quantitative dataset contributor
Masataka Yanagawa (RIKEN)
Image datasets
4
Quantitative datasets
0
Dataset List
Thumbnail
Dataset
Organism
Kind / Links
The tracjectories of EGFR-GFP from colocalization of EGFR and PI(4,5)P2 in living cells after EGF stimulation
The tracjectories of EGFR-GFP from colocalization of EGFR and PI(4,5)P2 in living cells before EGF stimulation
The tracjectories of JF549–PI(4,5)P2 from colocalization of EGFR and PIP2 in living cells after EGF stimulation
The tracjectories of JF549–PI(4,5)P2 from colocalization of EGFR and PIP2 in living cells before EGF stimulation