Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Recycling Endosomes and Transcytosis

Recycling Endosomes and Transcytosis

The recycling endosome, also known as the endosomal recycling compartment (ERC), is a part of the slow-recycling process of the endocytic pathway. Molecules internalized through receptor-mediated endocytosis are either degraded in the lysosomes or are recycled to the plasma membrane through the fast- or slow-recycling route.
The recycling endosome is not a single organelle but an extensively tubulated network of recycling pathways. It functions in storing molecules or transporting them across...

Coat Assembly and GTPases

Coat Assembly and GTPases

Vesicles incorporate different coat protein subunits in different cell locations, which changes the properties of the coat, such as the shape and geometry of the transport vesicles. Thus, vesicle coat proteins also play a significant role in cargo selection.
Coat assembly depends on the local availability of phosphatidylinositol phosphates or PIPs and GTP-binding proteins. Adaptor proteins, which link the coat proteins to the membrane, bind to these PIPs and play a crucial role in controlling...

The Early Endosome: Endocytosis of Transferrin

The Early Endosome: Endocytosis of Transferrin

Essential proteins such as insulin or low-density lipoprotein (LDL) and micronutrients such as iron enter a eukaryotic cell through receptor-mediated endocytosis. Subsequently, the early endosomes fuse with the vesicles containing such receptor-ligand complexes and play a vital role in sorting the incoming ligands and receptors. While the ligands are either degraded inside the vesicle or released into the cytosol, their receptors are returned to the plasma membrane for further rounds of...

Maturation of Endosomes

Maturation of Endosomes

The early endosome containing internalized molecules matures through transformations in its location, morphology, intraluminal pH, and membrane protein composition. Together, these changes result in a more acidic late endosome that contains multiple intraluminal vesicles; therefore, the late endosome is also called a multivesicular body (MVB).
Changes in location
The maturing endosome moves along microtubules from the periphery of the cell towards the perinuclear region. This movement of the...

Clathrin Coated Vesicles

Clathrin Coated Vesicles

Clathrin-coated vesicles use endocytosis to transport receptors and lysosomal hydrolases from the Golgi to the lysosome in the late secretory pathway. Clathrin-mediated endocytosis was the first described endocytic process, and Clathrin-coated vesicles remain one of the most well-studied transport vesicles. The molecular machinery that generates clathrin-coated vesicles comprises over 50 proteins that precisely coordinate vesicle formation. Cell surface receptors concentrated in indented sites...

Export of Misfolded Proteins out of the ER

Export of Misfolded Proteins out of the ER

After folding, the ER assesses the quality of secretory and membrane proteins. The correctly folded proteins are cleared by the calnexin cycle for transport to their final destination, while misfolded proteins are held back in the ER lumen. The ER chaperones attempt to unfold and refold the misfolded proteins but sometimes fail to achieve the correct native conformation. Such terminally misfolded proteins are then exported to the cytosol by ER-associated degradation or ERAD pathway for...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Divergent Rickettsia species exhibit distinct mechanisms of actin-based motility.

The Journal of cell biology·2026

Same author

Correction: Interferon receptor-deficient mice are susceptible to eschar-associated rickettsiosis.

eLife·2025

Same author

Divergent <i>Rickettsia</i> species exhibit distinct mechanisms of actin-based motility.

bioRxiv : the preprint server for biology·2025

Same author

The <i>Rickettsia</i> actin-based motility effectors RickA and Sca2 contribute differently to cell-to-cell spread and pathogenicity.

mBio·2025

Same author

From pipette to palette: Bridging science and art at <i>MBoC</i>.

Molecular biology of the cell·2024

Same author

Cryo-electron tomography of stationary phase <i>Burkholderia thailandensis</i>.

microPublication biology·2024

Search research articles

Related Experiment Video

Updated: Jun 16, 2026

The Microscopy-Based Assay to Study and Analyze the Recycling Endosomes using SNARE Trafficking

The Microscopy-Based Assay to Study and Analyze the Recycling Endosomes using SNARE Trafficking

Published on: February 12, 2022

WASH and the Arp2/3 complex regulate endosome shape and trafficking.

Steve N Duleh¹, Matthew D Welch

¹Department of Molecular and Cell Biology, University of California-Berkeley, Berkeley, CA 94720, USA.

Cytoskeleton (Hoboken, N.J.)

|February 23, 2010

Summary

The WASH protein activates the Arp2/3 complex, promoting actin assembly crucial for endosome shape and function. This study reveals WASH

More Related Videos

In Vitro Polymerization of F-actin on Early Endosomes

In Vitro Polymerization of F-actin on Early Endosomes

Published on: August 28, 2017

Density Gradient Ultracentrifugation for Investigating Endocytic Recycling in Mammalian Cells

Density Gradient Ultracentrifugation for Investigating Endocytic Recycling in Mammalian Cells

Published on: June 30, 2021

Related Experiment Videos

Last Updated: Jun 16, 2026

The Microscopy-Based Assay to Study and Analyze the Recycling Endosomes using SNARE Trafficking

The Microscopy-Based Assay to Study and Analyze the Recycling Endosomes using SNARE Trafficking

Published on: February 12, 2022

In Vitro Polymerization of F-actin on Early Endosomes

In Vitro Polymerization of F-actin on Early Endosomes

Published on: August 28, 2017

Density Gradient Ultracentrifugation for Investigating Endocytic Recycling in Mammalian Cells

Density Gradient Ultracentrifugation for Investigating Endocytic Recycling in Mammalian Cells

Published on: June 30, 2021

Area of Science:

Cell Biology
Molecular Biology
Biochemistry

Background:

Nucleation-promoting factors (NPFs) regulate actin assembly, a fundamental cellular process.
Different NPFs are involved in distinct cellular functions like endocytosis, cell migration, and intracellular transport.
The specific roles and localization of the WASH protein, an NPF, remained largely uncharacterized.

Purpose of the Study:

To investigate the cellular localization and function of the WASH protein.
To determine the role of WASH in regulating the Arp2/3 complex and actin assembly.
To elucidate the impact of WASH-mediated actin dynamics on endosomal trafficking.

Main Methods:

In vitro biochemical assays to assess Arp2/3 complex activation by WASH.
RNA interference (RNAi) to silence WASH and Arp2/3 complex expression in human cells.
Drug treatments to disrupt actin polymerization.
Confocal microscopy to visualize endosomal compartments (Rab5, Rab11, LAMP1) and actin filaments.

Main Results:

Human WASH directly activated the Arp2/3 complex in vitro and in cells, independent of autoinhibition.
WASH localized to Rab5- and Rab11-positive endosomes, which were enriched in actin filaments.
Silencing WASH or Arp2/3, or disrupting actin, led to enlarged and elongated endosomes.
WASH depletion and actin disruption impaired the transport of epidermal growth factor (EGF) to late endosomes.

Conclusions:

WASH is a potent activator of the Arp2/3 complex, likely regulated by interacting proteins.
WASH-mediated actin polymerization is essential for maintaining endosome morphology and maturation.
This study uncovers a novel role for WASH and the Arp2/3 complex in the degradative pathway of endocytic trafficking.