Related Experiment Video
Updated: Oct 6, 2025

The Soft Agar Colony Formation Assay
Published on: October 27, 2014
FBXW7-mediated ERK3 degradation regulates the proliferation of lung cancer cells
Hyun-Jung An1,2, Cheol-Jung Lee1,3, Ga-Eun Lee1,2
1College of Pharmacy, The Catholic University of Korea, 43, Jibong-Ro, Wonmi-Gu, Bucheon-si, Gyeonggi-Do, 14662, Republic of Korea.
Abstract:
Extracellular signal-regulated kinase 3 (ERK3) is an atypical member of the mitogen-activated protein kinase (MAPK) family, members of which play essential roles in diverse cellular processes during carcinogenesis, including cell proliferation, differentiation, migration, and invasion. Unlike other MAPKs, ERK3 is an unstable protein with a short half-life. Although deubiquitination of ERK3 has been suggested to regulate the activity, its ubiquitination has not been described in the literature. Here, we report that FBXW7 (F-box and WD repeat domain-containing 7) acts as a ubiquitination E3 ligase for ERK3. Mammalian two-hybrid assay and immunoprecipitation results demonstrated that ERK3 is a novel binding partner of FBXW7. Furthermore, complex formation between ERK3 and the S-phase kinase-associated protein 1 (SKP1)-cullin 1-F-box protein (SCF) E3 ligase resulted in the destabilization of ERK3 via a ubiquitination-mediated proteasomal degradation pathway, and FBXW7 depletion restored ERK3 protein levels by inhibiting this ubiquitination. The interaction between ERK3 and FBXW7 was driven by binding between the C34D of ERK3, especially at Thr417 and Thr421, and the WD40 domain of FBXW7. A double mutant of ERK3 (Thr417 and Thr421 to alanine) abrogated FBXW7-mediated ubiquitination. Importantly, ERK3 knockdown inhibited the proliferation of lung cancer cells by regulating the G1/S-phase transition of the cell cycle. These results show that FBXW7-mediated ERK3 destabilization suppresses lung cancer cell proliferation in vitro.
Insights
FBXW7 targets ERK3 for ubiquitination and degradation, suppressing lung cancer cell proliferation by controlling cell cycle progression. This discovery reveals a new mechanism in cancer biology.
Area of Science:
- Molecular Biology
- Cell Biology
- Cancer Research
Background:
- Extracellular signal-regulated kinase 3 (ERK3) is an atypical MAPK involved in cell processes crucial for cancer.
- ERK3 is unstable, and its regulation by ubiquitination has not been previously described.
- Mitogen-activated protein kinases (MAPKs) are key regulators in cell proliferation, differentiation, migration, and invasion during carcinogenesis.
Purpose of the Study:
- To identify the E3 ligase responsible for ERK3 ubiquitination.
- To elucidate the mechanism by which FBXW7 regulates ERK3 stability.
- To investigate the role of ERK3 in lung cancer cell proliferation.
Main Methods:
- Mammalian two-hybrid assay and immunoprecipitation to confirm protein interactions.
- Ubiquitination assays and Western blotting to assess protein stability and degradation.
- ERK3 knockdown and cell cycle analysis (G1/S-phase transition) in lung cancer cells.
Main Results:
- FBXW7 was identified as the E3 ligase that ubiquitylates ERK3.
- ERK3 forms a complex with the SCF-FBXW7 E3 ligase, leading to ERK3 ubiquitination and proteasomal degradation.
- FBXW7 depletion stabilized ERK3 by inhibiting its ubiquitination.
- ERK3 knockdown suppressed lung cancer cell proliferation by inhibiting G1/S-phase transition.
Conclusions:
- FBXW7-mediated ubiquitination and degradation of ERK3 is a novel mechanism for regulating ERK3 stability.
- FBXW7-mediated destabilization of ERK3 suppresses lung cancer cell proliferation.
- Targeting the FBXW7-ERK3 interaction may offer a therapeutic strategy for lung cancer.
More Related Videos
06:51Utilizing 18F-FDG PET/CT Imaging and Quantitative Histology to Measure Dynamic Changes in the Glucose Metabolism in Mouse Models of Lung Cancer
Published on: July 21, 2018
11:32Identification of Transcription Factor Regulators using Medium-Throughput Screening of Arrayed Libraries and a Dual-Luciferase-Based Reporter
Published on: March 27, 2020
Related Concept Videos
Mitogens and the Cell Cycle
Receptor Downregulation in MVBs
The EGFR can initiate signaling pathways that lead to cell proliferation, migration, and differentiation. Overexpression of EGFR stimulates cells to proliferate. Excessive EGFR...
Abnormal Proliferation
Negative Regulator Molecules
MAPK Signaling Cascades
PI3K/mTOR/AKT Signaling Pathway