Related Experiment Video

Updated: Jun 4, 2026

Production of Pseudotyped Particles to Study Highly Pathogenic Coronaviruses in a Biosafety Level 2 Setting
08:40

Production of Pseudotyped Particles to Study Highly Pathogenic Coronaviruses in a Biosafety Level 2 Setting

Published on: March 1, 2019

Mobility and interactions of coronavirus nonstructural protein 4

Marne C Hagemeijer1, Mustafa Ulasli, Annelotte M Vonk

  • 1Virology Division, Department of Infectious Diseases & Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, Netherlands.

Journal of Virology
|February 25, 2011
PubMed

Insights

Mouse hepatitis coronavirus nonstructural protein 4 (nsp4) localizes to the endoplasmic reticulum and viral replication sites. Nsp4 mobility is restricted at replication sites, where it interacts with other viral proteins like nsp3 and nsp6.

Area of Science:

  • Virology
  • Cell Biology
  • Molecular Biology

Background:

  • Mouse hepatitis coronavirus (MHV) is a significant pathogen.
  • Understanding coronavirus replication mechanisms is crucial for developing antivirals.
  • Nonstructural protein 4 (nsp4) is essential for MHV replication.

Purpose of the Study:

  • To investigate the localization and dynamics of MHV nsp4.
  • To explore the interactions of nsp4 with other viral proteins.
  • To elucidate the role of nsp4 in the formation of coronavirus replication structures.

Main Methods:

  • Green fluorescent protein (GFP) tagging of nsp4.
  • Confocal microscopy to observe protein localization.
  • Fluorescence loss in photobleaching (FLIP) and fluorescence recovery after photobleaching (FRAP) experiments to assess protein mobility.
  • Coexpression studies to analyze protein-protein interactions.

Main Results:

  • GFP-tagged nsp4 localized to the endoplasmic reticulum (ER) and coronavirus replicative structures.
  • ER membranes are continuous with membranes of replicative structures.
  • Nsp4 mobility was restricted at replicative structures compared to the ER.
  • Nsp4 engaged in homotypic and heterotypic interactions, notably with nsp3 and nsp6.
  • Coexpression of nsp4 and nsp3 altered their subcellular localization.

Conclusions:

  • Nsp4 plays a key role in the organization of coronavirus replication sites.
  • The restricted mobility and interactions of nsp4 suggest its involvement in membrane remodeling.
  • Nsp4 interacts with nsp3 and nsp6, indicating a coordinated function in viral replication.

More Related Videos

Engineering Antiviral Agents via Surface Plasmon Resonance13:00

Engineering Antiviral Agents via Surface Plasmon Resonance

Published on: June 14, 2022

Related Concept Videos

Coronavirus01:29

Coronavirus

Coronaviruses, including the severe acute respiratory syndrome coronavirus (SARS-CoV), are enveloped viruses characterized by their single-stranded, positive-sense RNA genome and helical nucleocapsid structure. The hallmark of these viruses is their club-shaped spike (S) glycoproteins that protrude from the viral envelope, facilitating attachment to host cells. Typically, coronaviruses infect the upper respiratory tract, often causing mild or asymptomatic disease. However, certain strains like...
Intracellular Movement of Viruses and Bacteria01:10

Intracellular Movement of Viruses and Bacteria

Intracellular bacteria and viruses often comprise a group of highly infectious pathogens that can cause several diseases. Bacterial pathogens include those belonging to the genus Rickettsia responsible for conditions such as rocky mountain spotted fever and the Mediterranean spotted fever; Chlamydia, a genus responsible for a sexually transmitted disease; Coxiella burnetii, an agent responsible for Q fever. Viral pathogens include vaccinia—a poxvirus, and herpes simplex virus—a virus that...
Inhibitors Of Virion Release01:25

Inhibitors Of Virion Release

Viral replication and dissemination rely on efficient mechanisms for host cell entry, genome replication, assembly, and release. Influenza viruses, such as types A and B, are negative-sense single-stranded RNA viruses with a segmented genome, that depend on two critical surface glycoproteins to carry out these processes: hemagglutinin (HA) and neuraminidase (NA). HA initiates infection by binding to sialic acid residues on the surface of host epithelial cells, facilitating receptor-mediated...
Conjugated Proteins02:50

Conjugated Proteins

Simple proteins and protein complexes contain only amino acids. In contrast, many other proteins, called conjugated proteins, covalently bond with non-protein moieties.
Nucleoproteins are protein complexes that contain nucleic acids, categorized as deoxyribonucleoproteins (DNPs) or ribonucleoproteins (RNPs) respectively. The nucleosome is a typical example of a DNP where nuclear DNA is associated with histone proteins. The major antigen for the Covid-19 virus SARS-CoV is an RNP that is critical...
Inhibitors of Virion Maturation and Assembly01:19

Inhibitors of Virion Maturation and Assembly

As part of their replication cycle, certain viruses synthesize long precursor proteins called polyproteins within infected host cells. In human immunodeficiency virus (HIV), two major polyproteins are produced: Gag and Gag-Pol. The Gag polyprotein supplies the structural components of the virus, while Gag-Pol includes essential viral enzymes such as reverse transcriptase, integrase, and protease. After synthesis, these polyproteins move to the host cell membrane, where they assemble into an...
Respiratory Syncytial Virus Disease01:29

Respiratory Syncytial Virus Disease

Human respiratory syncytial virus (RSV) is a widespread pathogen that primarily targets infants and young children but also poses a serious health risk to elderly and immunocompromised individuals. Belonging to the Pneumoviridae family, RSV is a negative-sense, single-stranded RNA virus within the Pneumovirus genus. Its global health burden is significant, with millions of cases annually resulting in hospitalizations and mortality, particularly in resource-limited settings. Although most...