SSBD:database

Project ID

439-Takayama-dynamicsEGFR

Project SID

439

Title

EGFR dynamics on CHO–K1 cells with depletion and supplementation of cholesterol

Description

Epidermal growth factor receptor (EGFR)-mediated signal transduction controls cell growth and proliferation. The signaling pathway is regulated so that it is activated only by external EGF stimuli, but the mechanisms that prevent EGF-independent spontaneous activation of EGFR-mediated signaling are unknown. Here the authors report cholesterol depletion activates EGFR-mediated signaling without EGF. They applied automated single-molecule imaging to EGFR and characterized the lateral diffusion and cluster formation on cholesterol-depleted and cholesterol-supplemented membranes. In cells in which cholesterol was depleted by methyl-beta-cyclodextrin treatment, EGFR exhibited a reduction in lateral diffusion, an acceleration of cluster formation, and autophosphorylation without EGF. Concurrently, extracellular signal-regulated kinase (ERK), which is regulated by EGFR-mediated signaling, exhibited phosphorylation and nuclear translocation without EGF. These cholesterol depletion-induced changes were similar, albeit less efficient, to those that occurred with EGF stimulation in normal cells without methyl-beta-cyclodextrin, indicating the spontaneous activation of EGFR signaling. The exogenous supplementation of cholesterol suppressed the methyl-beta-cyclodextrin-induced spontaneous activation of EGFR and ERK nuclear translocation. Single-molecule imaging of EGFR in a large number of cells revealed cell-to-cell heterogeneity, with a sub-population showing a high ability for spontaneous activation. These results provide evidence that EGFR-mediated signaling is properly regulated by cholesterol metabolism to prevent uncontrolled spontaneous activation.

Submission Date

2025-06-09

Opened Date

-

Release Date

2025-11-28

Update Date

-

Kind

License

Project URL

-

Project DOI

-

Metadata Version

-

Template Version

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Funding Information

AMED Grant Number JP22ym0126815, Japan; JST CREST Grant Number JPMJCR21E1, Japan; MEXT Grants-in-Aid for Scientific Research(B) (18H01839, 22H02593), Japan; JSPS Grant-in-Aid for Challenging Research (Pioneering) (22K18363DSLM), Japan.

Total

36 files / 1.4 GB

Image datasets

zipped 8 files / 199.4 MB, raw image 8 files / 506.3 MB, total 16 files / 705.7 MB

Quantitative datasets

zipped 0 files / 0 bytes, raw bdml 0 files / 0 bytes, total 0 files / 0 bytes

Organism

Strain

-

Cell

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Cell line

MeSH

Anatomical Entity

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UBERON

Biological Process

Cellular Component

Molecular Function

Medical Subject Headings

Method involved in biological imaging