Related Experiment Video

Updated: Aug 4, 2026

RhoC GTPase Activation Assay
09:58

RhoC GTPase Activation Assay

Published on: August 22, 2010

Motility and invasion are differentially modulated by Rho family GTPases

J Banyard1, B Anand-Apte, M Symons

  • 1Department of Surgical Research, Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.

Oncogene
|March 4, 2000
PubMed

Insights

Cell invasion through 3D matrices requires Cdc42, Rac, and Rho GTPase activity. Optimal activation levels of these GTPases are crucial for cell invasion, with migration and invasion being differentially regulated.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Cell migration in vivo often involves tissue matrix invasion.
  • Signaling pathways regulating cell invasion through 3D matrices are not well understood.
  • Small GTPases Cdc42, Rac, and Rho are critical regulators of the actin cytoskeleton and cell adhesion.

Purpose of the Study:

  • To investigate the roles of Cdc42, Rac, and Rho GTPases in PDGF-BB-stimulated cell invasion through 3D collagen matrices.
  • To differentiate the roles of these GTPases in 3D invasion versus 2D migration.

Main Methods:

  • Expression of dominant-negative and constitutively active forms of Cdc42, Rac, and Rho in Rat1 fibroblasts.
  • Assessment of cell invasion into 3D collagen matrices.
  • Assessment of cell locomotion across a planar substrate using a Boyden chamber assay.
  • Investigation of the role of PI-3-kinase activity.

Main Results:

  • Inhibition of Cdc42, Rac, or Rho GTPase activity blocked PDGF-BB-stimulated invasion into 3D matrices.
  • Only Rac activation was essential for PDGF-BB-stimulated 2D migration.
  • Constitutively active Cdc42 or Rho, and to a lesser extent Rac, inhibited PDGF-induced invasion.
  • Constitutively active Rac stimulated basal invasion independently of PI-3-kinase, an effect dependent on its effector function.

Conclusions:

  • Cellular invasion through 3D matrices necessitates optimal activation levels of Cdc42, Rho, and Rac GTPases.
  • Cell migration and invasion are differentially regulated by Rho family GTPases.
  • These findings provide insight into the molecular mechanisms governing cell invasion in complex 3D environments.

More Related Videos

Comparing the Affinity of GTPase-binding Proteins using Competition Assays10:37

Comparing the Affinity of GTPase-binding Proteins using Competition Assays

Published on: October 8, 2015

Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay13:51

Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay

Published on: November 11, 2018

Related Concept Videos

Cell Migration01:09

Cell Migration

Cell migration, the process by which cells move from one location to another, is essential for the proper development and viability of organisms throughout their life. When cells are not able to migrate properly to their ordained locations, various disorders may occur. For example, disruption in cell migration causes chronic inflammatory diseases such as arthritis.
Cancer Cell Migration through Invadopodia01:35

Cancer Cell Migration through Invadopodia

Invadosome is a broad category of cell surface structures with proteolytic activity that  degrades the extracellular matrix (ECM). Invadosomes are present in normal cell types, including macrophages, endothelial cells, and neurons, as well as tumor cells. Although the macrophage podosomes and tumor cell invadopodia are classified as invadosomes, they have different structures, molecular pathways, and functions. Podosomes are short structures that last for a few minutes. However, invadopodia can...
Role of Myosin in Cell Migration01:18

Role of Myosin in Cell Migration

Myosins are multimeric motor proteins involved in various cellular processes such as migration, adhesion, and proliferation. Myosin II is the most common type in animal cells, which binds and cross-links actin filaments.
Myosin II  is a hexamer comprising two heavy chains with globular heads and coiled-coil tails, two regulatory light chains, and two essential light chains. The ATPase sites on the myosin heads hydrolyze ATP, and the released phosphate generates the force for contraction. It is...
Cell Polarization by Rho Proteins01:21

Cell Polarization by Rho Proteins

Cell polarity is the asymmetric distribution of cellular and membrane components, making one side of the cell different from the other. This polarity is essential to many processes such as embryogenesis, axon migration, glucose transport across epithelial cells, and directional cell migration. A migrating cell responds to intracellular or extracellular signals via molecular cascades that reorganize the actin cytoskeleton to establish this polarity. In these cells, the Rho family proteins Cdc42,...
Cytoskeletal Coordination in Cell Migration01:32

Cytoskeletal Coordination in Cell Migration

A migrating cell changes its shape during the cyclic events of attachment and detachment from the substratum and repositions the cell organelles correspondingly. These complex events are orchestrated by the dynamic cytoskeletal network comprising actin filaments, intermediate filaments, and microtubules. Cytoskeletal crosstalk — the direct and indirect communication between the different components — is crucial for this coordination. Direct communication involves various linker proteins that...
Small GTPases - Ras and Rho01:24

Small GTPases - Ras and Rho

Ras and Rho are small monomeric GTPases that act downstream of receptor tyrosine kinase (RTK) and regulate various cellular processes. These GTPases switch between active and inactive states by binding to guanine nucleotides.
Three regulatory proteins control their activity: