Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Molecular Chaperones and Protein Folding03:00

Molecular Chaperones and Protein Folding

17.7K
The native conformation of a protein is formed by interactions between the side chains of its constituent amino acids. When the amino acids cannot form these interactions, the protein cannot fold by itself and needs chaperones. Notably, chaperones do not relay any additional information required for the folding of polypeptides; the native conformation of a protein is determined solely by its amino acid sequence. Chaperones catalyze protein folding without being a part of the folded protein.
The...
17.7K
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

5.7K
DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...
5.7K
Pinching-off of Coated Vesicles01:32

Pinching-off of Coated Vesicles

3.1K
Vesicle budding is orchestrated by distinct cytosolic proteins such as adaptor proteins, coat proteins, and GTPases. To initiate vesicle budding, membrane-bending proteins containing crescent-shaped BAR domains bind to the lipid heads in the bilayer and distort the membrane to form a protein-coated vesicle bud. Adaptors proteins such as AP2 for clathrin-coated vesicles can nucleate on the deformed membrane. Finally, coat proteins such as clathrin or COPI and COPII assemble into a coat forming...
3.1K
Export of Misfolded Proteins out of the ER01:32

Export of Misfolded Proteins out of the ER

3.5K
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...
3.5K
Synthesis of Phosphatidylcholine in the ER Membrane01:27

Synthesis of Phosphatidylcholine in the ER Membrane

3.0K
The ER synthesizes lipids for building cell membranes and performing cellular functions such as energy storage and signaling. The lipid synthesis machinery embedded in the ER membrane primarily collects all reactants from the cytosol. Following synthesis, the secretory pathway and the ER contact sites distribute these lipids to other cellular organelles. Additionally, the energy-rich triacylglycerides are transported from the ER via lipid droplets.
The major components of all eukaryotic cell...
3.0K
Translocation of Proteins into the Mitochondria01:19

Translocation of Proteins into the Mitochondria

3.0K
Mitochondrial precursors are translocated to the internal subcompartments via independent mechanisms involving distinct protein machineries called translocases.
Sorting of outer membrane proteins:
Mitochondrial outer membrane proteins are of two types: the transmembrane, beta-barrel porins, and the membrane-anchored, alpha-helical proteins. Beta-barrel porin precursors are translocated by the TOM complex and inserted into the outer mitochondrial membrane by the SAM complex. In contrast,...
3.0K

You might also read

Related Articles

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

Sort by
Same author

Association-induced folding governs surrogate light chain and pre-B cell receptor core assembly.

Nature communications·2026
Same author

The making of multispecific immunoglobulins - a clinical perspective.

mAbs·2026
Same author

Evolutionary conservation of ubiquitin-like protein urmylation as revealed by URM1 gene shuffle from archaea to yeast.

Communications biology·2025
Same author

Improved targeted delivery of antisense oligonucleotide with an antibody mask.

Nucleic acids research·2025
Same author

Autophosphorylation of conserved yeast and human casein kinase 1 isozymes regulates Elongator-dependent tRNA modifications.

Nucleic acids research·2025
Same author

Stress response pathways: machineries and mechanisms.

Biological chemistry·2025

Related Experiment Video

Updated: Jun 1, 2025

Monitoring eIF4F Assembly by Measuring eIF4E-eIF4G Interaction in Live Cells
08:47

Monitoring eIF4F Assembly by Measuring eIF4E-eIF4G Interaction in Live Cells

Published on: May 1, 2020

3.0K

Diphthamide synthesis is linked to the eEF2-client chaperone machinery.

Lars Kaduhr1, Klaus Mayer2, Raffael Schaffrath1

  • 1Department of Microbiology, Kassel University, Germany.

FEBS Letters
|January 18, 2025
PubMed
Summary

The diphthamide modification of eukaryotic translation elongation factor 2 (eEF2) is crucial for protein synthesis. Yeast studies reveal that Hsp90 co-chaperones Hgh1 and Cpr7 are essential for this modification, ensuring accurate protein production.

Keywords:
Cpr7Hgh1J‐proteinchaperonediphthamideelongation factor 2 (eEF2)

More Related Videos

In Situ Monitoring of Transiently Formed Molecular Chaperone Assemblies in Bacteria, Yeast, and Human Cells
00:08

In Situ Monitoring of Transiently Formed Molecular Chaperone Assemblies in Bacteria, Yeast, and Human Cells

Published on: September 2, 2019

7.0K
Detection of the pH-dependent Activity of Escherichia coli Chaperone HdeB In Vitro and In Vivo
08:32

Detection of the pH-dependent Activity of Escherichia coli Chaperone HdeB In Vitro and In Vivo

Published on: October 23, 2016

10.5K

Related Experiment Videos

Last Updated: Jun 1, 2025

Monitoring eIF4F Assembly by Measuring eIF4E-eIF4G Interaction in Live Cells
08:47

Monitoring eIF4F Assembly by Measuring eIF4E-eIF4G Interaction in Live Cells

Published on: May 1, 2020

3.0K
In Situ Monitoring of Transiently Formed Molecular Chaperone Assemblies in Bacteria, Yeast, and Human Cells
00:08

In Situ Monitoring of Transiently Formed Molecular Chaperone Assemblies in Bacteria, Yeast, and Human Cells

Published on: September 2, 2019

7.0K
Detection of the pH-dependent Activity of Escherichia coli Chaperone HdeB In Vitro and In Vivo
08:32

Detection of the pH-dependent Activity of Escherichia coli Chaperone HdeB In Vitro and In Vivo

Published on: October 23, 2016

10.5K

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • The diphthamide modification of eukaryotic translation elongation factor 2 (eEF2) is vital for accurate protein synthesis.
  • While enzymes responsible for diphthamide synthesis are identified, the coordination between eEF2 synthesis and its modification remains unclear.

Purpose of the Study:

  • To investigate the coordination of eEF2 synthesis and diphthamide modification.
  • To identify factors involved in ensuring only modified eEF2 is maintained.

Main Methods:

  • Analysis of physical and genetic interactions from the BioGRID database in yeast.
  • Examination of eEF2 modification in deletion strains of Hsp90 co-chaperones (Hgh1 and Cpr7).

Main Results:

  • Physical and genetic interactions link diphthamide synthesis enzymes with chaperones, including Hsp90 co-chaperones Hgh1 and Cpr7.
  • Deletion of Hgh1 or Cpr7 resulted in eEF2 lacking the diphthamide modification.
  • Defects in other co-chaperones did not affect eEF2 diphthamide modification.

Conclusions:

  • Diphthamide synthesis requires not only Dph enzymes but also eEF2-interacting co-chaperones Hgh1 and Cpr7.
  • The process likely necessitates a specific conformational state of eEF2 maintained by particular chaperones.