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Rab Cascades

Rab Cascades

Rab GTPases act in a regulated cascade during membrane fusion, helping the lipid bilayers mix. The Rab family of proteins are active when bound to GTP, and inactive when bound to GDP. Hence, they act as guanine nucleotide-dependent molecular switches. Rab-GTP recognizes and binds to long or short-range tethering proteins to capture the target vesicle. These tethers coordinate with SNAREs on the vesicle and the target membrane to assemble the trans SNARE complex that locks the mixing bilayers.

Rab Proteins

Rab Proteins

Rab proteins constitute the largest family of monomeric GTPases, of which 70 members are present in humans. Rab proteins and their effectors regulate consecutive stages of vesicle transport such as vesicle transport, docking, and fusion to the correct recipient membrane.
Rab proteins switch between a cytosolic, GDP-bound inactive state and a membrane-anchored, GTP-bound active state. By themselves, Rabs show slow rates of GDP/GTP exchange and GTP hydrolysis. Thus, Rab proteins are considered...

Hepatitis

Hepatitis

Hepatitis is an inflammatory condition of the liver most commonly caused by hepatotropic viruses (A–E), though non-infectious causes such as alcohol and drugs also exist.Hepatitis AHepatitis A virus (HAV) is a non-enveloped RNA virus of the Picornaviridae family. It is primarily transmitted via the fecal-oral route, typically through ingestion of contaminated food or water. After ingestion, HAV enters the bloodstream through the oropharynx or intestinal epithelium and reaches the liver. The...

SNAREs and Membrane Fusion

SNAREs and Membrane Fusion

Once a transport vesicle has recognized its target organelle, the vesicular membrane needs to fuse with the target membrane to unload the cargo. Transmembrane proteins called SNAREs present on organelle membranes and their vesicles, mediate vesicle fusion.
SNAREs exist in pairs that symmetrically interact and catalyze the fusion of the lipid bilayers in vesicle and target organelle. v-SNARE in the vesicle membrane are single polypeptide chains that bind to a complementary t-SNARE, composed of 2...

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...

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Related Experiment Video

Updated: Jun 17, 2026

A Protocol for Analyzing Hepatitis C Virus Replication

A Protocol for Analyzing Hepatitis C Virus Replication

Published on: June 26, 2014

Endocytic Rab proteins are required for hepatitis C virus replication complex formation.

David Manna¹, Jason Aligo, Chenjia Xu

¹Department of Biochemistry and Molecular Biology, The Pennsylvania State University, 308 Althouse Laboratory, University Park, PA 16802, USA.

|December 17, 2009

Summary

Hepatitis C virus (HCV) NS4B protein foci formation is crucial for viral replication. This study reveals that endocytic proteins Rab5 and Rab7 facilitate NS4B foci assembly, impacting HCV RNA synthesis.

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Last Updated: Jun 17, 2026

A Protocol for Analyzing Hepatitis C Virus Replication

A Protocol for Analyzing Hepatitis C Virus Replication

Published on: June 26, 2014

Two Methods of Heterokaryon Formation to Discover HCV Restriction Factors

Two Methods of Heterokaryon Formation to Discover HCV Restriction Factors

Published on: July 16, 2012

Lipid Droplet Isolation for Quantitative Mass Spectrometry Analysis

Lipid Droplet Isolation for Quantitative Mass Spectrometry Analysis

Published on: April 17, 2017

Area of Science:

Virology
Cell Biology
Molecular Biology

Background:

Hepatitis C virus (HCV) infection involves NS4B protein remodeling host membranes.
NS4B protein forms replication complexes (RCs) appearing as foci under fluorescence microscopy (FM).
The role of Rab proteins in NS4B foci formation remains unclear.

Purpose of the Study:

To investigate the involvement of Rab proteins in the formation of NS4B foci.
To elucidate the functional significance of Rab proteins in HCV replication.

Main Methods:

Biochemical fractionation of HCV replicon-expressing cells.
Fluorescence microscopy (FM) to visualize NS4B foci and Rab protein association.
Dominant-negative (DN) Rab protein expression and gene silencing (siRNA) to assess functional impact.
Quantification of HCV RNA synthesis.

Main Results:

Isolated NS4B-bound fractions were competent for HCV RNA synthesis.
NS4B foci were selectively associated with Rab5 and Rab7 proteins.
Dominant-negative Rab6 impaired Rab5 recruitment to NS4B foci.
Silencing Rab5 or Rab7 significantly decreased HCV genome replication.
Expression of DN Rab5 or Rab7 resulted in reticular NS4B distribution.

Conclusions:

Endocytic proteins Rab5 and Rab7 facilitate NS4B foci formation.
Rab5 and Rab7 play a crucial role in Hepatitis C virus genome replication.
NS4B foci formation is dependent on specific Rab proteins, influencing viral RNA synthesis.