Related Experiment Video

Updated: Jun 17, 2025

Studying TGF-β Signaling and TGF-β-induced Epithelial-to-mesenchymal Transition in Breast Cancer and Normal Cells
06:54

Studying TGF-β Signaling and TGF-β-induced Epithelial-to-mesenchymal Transition in Breast Cancer and Normal Cells

Published on: October 27, 2020

12.9K

Targeted dephosphorylation of SMAD3 as an approach to impede TGF-β signaling

Abigail Brewer1, Jin-Feng Zhao1, Rotimi Fasimoye1

  • 1Medical Research Council (MRC) Protein Phosphorylation & Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.

Iscience
|August 6, 2024
PubMed

Insights

Targeted dephosphorylation of phospho-SMAD3 using a novel molecule (BDPIC) effectively reduced SERPINE-1 transcription, offering a new way to control TGF-β signaling pathways in diseases like cancer and fibrosis.

Area of Science:

  • Cellular signaling
  • Molecular biology
  • Biochemistry

Background:

  • Transforming growth factor-β (TGF-β) signaling regulates crucial cellular functions.
  • Dysregulated TGF-β signaling is linked to diseases such as fibrosis and cancer.
  • Phosphorylation of SMAD2 and SMAD3 controls TGF-β transcriptional responses.

Purpose of the Study:

  • To investigate targeted dephosphorylation of phospho-SMAD3 as a strategy to inhibit TGF-β-induced transcription.
  • To develop and utilize a bifunctional molecule for targeted protein modification.
  • To assess the impact of phospho-SMAD3 dephosphorylation on SERPINE-1 transcription.

Main Methods:

  • Development of a bifunctional molecule, BDPIC (bromoTAG-dTAG proximity-inducing chimera).
  • Utilizing CRISPR-Cas9 technology to generate homozygous double knock-in A549 cells (PPM1H/SMAD3).
  • Employing BDPIC to induce proximity between bromoTAG-PPM1H and dTAG-SMAD3 for targeted dephosphorylation.

Main Results:

  • BDPIC successfully redirected phosphatases to dephosphorylate phospho-SMAD3.
  • Demonstrated robust dephosphorylation of dTAG-SMAD3 in engineered cells.
  • Observed a significant decrease in SERPINE-1 (PAI-1) transcription.

Conclusions:

  • Targeted dephosphorylation of phospho-proteins is a viable strategy for modulating cell signaling.
  • BDPIC provides a novel tool for rewiring TGF-β signaling pathways.
  • This approach holds potential for therapeutic interventions in TGF-β-related diseases.
Keywords:
BiochemistryBiochemistry methodsCell biology

More Related Videos

Live Cell Imaging of the TGF- β/Smad3 Signaling Pathway In Vitro and In Vivo Using an Adenovirus Reporter System11:06

Live Cell Imaging of the TGF- β/Smad3 Signaling Pathway In Vitro and In Vivo Using an Adenovirus Reporter System

Published on: July 30, 2018

10.6K
Visualization and Quantification of TGFβ/BMP/SMAD Signaling under Different Fluid Shear Stress Conditions using Proximity-Ligation-Assay11:38

Visualization and Quantification of TGFβ/BMP/SMAD Signaling under Different Fluid Shear Stress Conditions using Proximity-Ligation-Assay

Published on: September 14, 2021

3.4K

Related Concept Videos

TGF - β Signaling Pathway01:16

TGF - β Signaling Pathway

The TGF-β signaling pathway regulates cell growth, differentiation, adhesion, motility, and development. TGF-β ligands that induce TGF-β signaling are synthesized in their latent form. Several proteases or cell surface receptors such as integrins act upon the latent form, releasing the active ligand. There are three types of mammalian TGF-βs: (TGF-β1, TGF-β2, and TGF-β3) that bind as homodimers or heterodimers to TGF-β receptors. The TGF-β receptors...
7.3K
The JAK-STAT Signaling Pathway01:20

The JAK-STAT Signaling Pathway

Several cytokine receptors have tightly bound Janus kinase or JAK proteins attached at their cytosolic tail. Small signaling molecules such as cytokines, growth hormones, or prolactins bind to the cytokine receptors and initiate their dimerization. The dimerization brings the cytosolic JAKs together that trans-phosphorylate and activates each other. The activated JAKs now phosphorylate cytosolic tails of the cytokine receptors, which serve as binding sites for adaptor proteins such as  SH2...
8.7K
PI3K/mTOR/AKT Signaling Pathway01:22

PI3K/mTOR/AKT Signaling Pathway

The mammalian target of rapamycin  (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1  (mTORC1) and mTOR complex 2  (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast,  mTORC2 consists of a...
3.5K
Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

When a ligand binds to a cell-surface receptor, the receptor's intracellular domain changes shape, which may either activate its enzyme function or allow its binding to other molecules. The initial signal is amplified by most signal transduction pathways. This means that a single ligand molecule can activate multiple molecules of a downstream target. Proteins that relay a signal are most commonly phosphorylated at one or more sites, activating or inactivating the protein. Kinases catalyze...
8.4K