Description
Anionic lipid molecules, including phosphatidylinositol- 4,5- bisphosphate (PI(4,5)P2),
are implicated in the regulation of epidermal growth factor receptor (EGFR). However, the role of
the spatiotemporal dynamics of PI(4,5)P2 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)P2 nanodomains in the plasma membrane of HeLa cells using super- resolution
single- molecule microscopy. The EGFR and PI(4,5)P2 nanodomains aggregated before stimulation
with epidermal growth factor (EGF) through transient visits of EGFR to the PI(4,5)P2 nanodomains.
The degree of coaggregation decreased after EGF stimulation and depended on phospholipase
Cγ, the EGFR effector hydrolyzing PI(4,5)P2. Artificial reduction in the PI(4,5)P2 content of the
plasma membrane reduced both the dimerization and autophosphorylation of EGFR after stimula-
tion with EGF. Inhibition of PI(4,5)P2 hydrolysis after EGF stimulation decreased phosphorylation of
EGFR- Thr654. Thus, EGFR kinase activity and the density of PI(4,5)P2 around EGFR molecules were
found to be mutually regulated.
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).