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

Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

6.9K
Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....
6.9K
RNA Stability01:53

RNA Stability

33.8K
Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
33.8K
Nucleotide Excision Repair01:38

Nucleotide Excision Repair

3.7K
DNA Distortion and Damage
Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...
3.7K
Radical Reactivity: Steric Effects01:10

Radical Reactivity: Steric Effects

2.0K
The presence of electron-donating, electron-withdrawing, or conjugating groups adjacent to a radical center, imparts electronic stabilization to the radicals. Examples of such electronically-stabilized radicals are triphenylmethyl, tetramethylpiperidine‐N‐oxide, and 2,2‐diphenyl‐1‐picrylhydrazyl. These radicals are remarkably stable and are known as persistent radicals. Some of the persistent radicals can even be isolated and purified.
Along with electronic...
2.0K
Mismatch Repair01:20

Mismatch Repair

5.1K
Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
5.1K
mRNA Stability and Gene Expression02:51

mRNA Stability and Gene Expression

5.7K
The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
5.7K

You might also read

Related Articles

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

Sort by
Same author

Long-Term Analysis of NRG Oncology RTOG 0539: A Phase II Trial of Observation for Low-Risk Meningioma and Radiotherapy for Intermediate- and High-Risk Meningioma.

Journal of clinical oncology : official journal of the American Society of Clinical Oncology·2026
Same author

The backside β-turn is a key structural element of Rad6-family E2 ubiquitin-conjugating enzymes.

The Biochemical journal·2026
Same author

The Ki-67 proliferation index and recurrence risk of intracranial meningioma: a multicenter, retrospective cohort study of 5,050 patients.

Acta neurochirurgica·2026
Same author

Fractionated radiotherapy adjuvant to surgery of WHO-2 meningioma with and without gross total resection: a multicenter, retrospective cohort study of 1,452 patients.

Journal of neuro-oncology·2026
Same author

circPCMTD1: a protein-coding circular RNA that regulates DNA damage response in BCR/ABL1-positive leukemias.

Blood·2026
Same author

Targeting epigenetic regulators: In-silico discovery of natural inhibitors against histone demethylase KDM4C.

PloS one·2026

Related Experiment Video

Updated: Aug 27, 2025

A Scalable, Cell-Based Method for the Functional Assessment of Ube3a Variants
06:35

A Scalable, Cell-Based Method for the Functional Assessment of Ube3a Variants

Published on: October 10, 2022

2.1K

Mutations of Rad6 E2 ubiquitin-conjugating enzymes at alanine-126 in helix-3 affect ubiquitination activity and

Prakash K Shukla1, Dhiraj Sinha2, Andrew M Leng1

  • 1Department of Radiation Oncology and Huntsman Cancer Institute, University of Utah School of Medicine, Salt Lake City, Utah, USA.

The Journal of Biological Chemistry
|September 26, 2022
PubMed
Summary

Mutations in Rad6

Keywords:
E2 enzymesNMR spectroscopyUBE2AUBE2Benzyme catalysisenzyme structuremolecular dynamics simulationsrad6ubiquitin

More Related Videos

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta
10:27

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta

Published on: December 5, 2019

9.0K
Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A EYFP-CENP-A
09:02

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A EYFP-CENP-A

Published on: June 10, 2020

5.7K

Related Experiment Videos

Last Updated: Aug 27, 2025

A Scalable, Cell-Based Method for the Functional Assessment of Ube3a Variants
06:35

A Scalable, Cell-Based Method for the Functional Assessment of Ube3a Variants

Published on: October 10, 2022

2.1K
Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta
10:27

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta

Published on: December 5, 2019

9.0K
Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A EYFP-CENP-A
09:02

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A EYFP-CENP-A

Published on: June 10, 2020

5.7K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • Rad6 is a conserved E2 ubiquitin-conjugating enzyme vital for transcription, DNA repair, and protein degradation.
  • The role of Rad6's secondary structures in its functions is largely unknown.
  • A previously identified alanine-126 (A126) mutation in Rad6's helix-3 causes various cellular defects.

Purpose of the Study:

  • To investigate the structural and functional impact of helix-3 A126 mutations in Rad6 and its human homologs.
  • To elucidate the molecular mechanisms underlying ubiquitination defects caused by these mutations.

Main Methods:

  • Utilized a multidisciplinary approach including genetics, biochemistry, biophysics, and computational methods.
  • Employed molecular dynamics simulations and Nuclear Magnetic Resonance (NMR) spectroscopy.
  • Analyzed ubiquitination activity, protein stability, and structural integrity.

Main Results:

  • Helix-3 A126 mutations impair Rad6's ability to ubiquitinate target proteins without affecting E3 ligase interactions.
  • These mutations lead to local and global structural disorder in Rad6, reducing its stability.
  • Structural deformations and compromised ubiquitination activity were also observed in human homologs UBE2A and UBE2B.
  • The mutations were found to disrupt both ubiquitin charging and discharging steps.

Conclusions:

  • The conserved helix-3 is essential for maintaining the structural organization of Rad6's catalytic pocket.
  • Helix-3 integrity is critical for the enzymatic activity and overall biological functions of Rad6-family enzymes.
  • Understanding these structural-functional relationships provides insights into E2 ubiquitin-conjugating enzyme mechanisms.