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

General Transcription Factors01:30

General Transcription Factors

6.7K
Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
6.7K
Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

11.0K
Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
11.0K
Transcription Initiation01:47

Transcription Initiation

20.1K
Initiation is the first step of transcription in eukaryotes. Prokaryotic RNA Polymerase (RNAP) can bind to the template DNA and start transcribing. On the other hand, transcription in eukaryotes requires additional proteins, called transcription factors, to first bind to the promoter region in the DNA template. This binding helps recruit the specific RNAP that can assemble on the DNA and start transcription.
The promoters and enhancers and their accessory proteins allow tight regulation of...
20.1K

You might also read

Related Articles

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

Sort by
Same author

Shear-Induced CROSS (Cellular RedOx Spreading Shield) Assembly Sustains Neurotrophic Extracellular Vesicle Production for Functional Neural Networks.

Advanced functional materials·2026
Same author

CPSF73 activation and 3' RNA polymerase II pausing are lost during readthrough transcription after heat shock.

Cell reports·2026
Same author

Multi-omics and biochemical reconstitution reveal CDK7-dependent mechanisms controlling RNA polymerase II function at gene 5'- and 3' ends.

Cell reports·2025
Same author

Multi-omics and biochemical reconstitution reveal CDK7-dependent mechanisms controlling RNA polymerase II function at gene 5'- and 3'-ends.

bioRxiv : the preprint server for biology·2025
Same author

Biomaterials for Cell Manufacturing.

ACS macro letters·2024
Same author

Laboratory Management of Mammalian Hosts for <i>Ixodes scapularis</i> -Host-Pathogen Interaction Studies.

Comparative medicine·2024

Related Experiment Video

Updated: Jan 12, 2026

Single-Molecule Imaging of EWS-FLI1 Condensates Assembling on DNA
07:05

Single-Molecule Imaging of EWS-FLI1 Condensates Assembling on DNA

Published on: September 8, 2021

2.7K

Single-molecule Microscopy Reveals That TFIIE Subunits Dynamically Interact With Preinitiation Complexes in a Manner

Stephen R Archuleta1, Ryan C Miller1, Julia A Mirita1

  • 1Department of Biochemistry, University of Colorado Boulder, 596 UCB, Boulder CO 80309, USA.

Journal of Molecular Biology
|November 3, 2025
PubMed
Summary

General transcription factor TFIIE dynamically interacts with preinitiation complexes (PICs) during gene transcription. Its subunits, TFIIEα and TFIIEβ, exhibit asynchronous behavior without TFIIH, and mutations impact their stability and kinetics.

Keywords:
RNA polymerase IITFIIEkineticssingle molecule microscopytranscription

More Related Videos

Visualizing Actin and Microtubule Coupling Dynamics In Vitro by Total Internal Reflection Fluorescence TIRF Microscopy
08:44

Visualizing Actin and Microtubule Coupling Dynamics In Vitro by Total Internal Reflection Fluorescence TIRF Microscopy

Published on: July 20, 2022

3.9K
Author Spotlight: Evaluation of Protein-Condensate Dynamics in Live Human Cells
06:48

Author Spotlight: Evaluation of Protein-Condensate Dynamics in Live Human Cells

Published on: January 5, 2024

5.1K

Related Experiment Videos

Last Updated: Jan 12, 2026

Single-Molecule Imaging of EWS-FLI1 Condensates Assembling on DNA
07:05

Single-Molecule Imaging of EWS-FLI1 Condensates Assembling on DNA

Published on: September 8, 2021

2.7K
Visualizing Actin and Microtubule Coupling Dynamics In Vitro by Total Internal Reflection Fluorescence TIRF Microscopy
08:44

Visualizing Actin and Microtubule Coupling Dynamics In Vitro by Total Internal Reflection Fluorescence TIRF Microscopy

Published on: July 20, 2022

3.9K
Author Spotlight: Evaluation of Protein-Condensate Dynamics in Live Human Cells
06:48

Author Spotlight: Evaluation of Protein-Condensate Dynamics in Live Human Cells

Published on: January 5, 2024

5.1K

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Transcription by RNA polymerase II (Pol II) necessitates general transcription factors (GTFs) to form preinitiation complexes (PICs) at gene promoters.
  • TFIIE is a crucial GTF that recruits TFIIH, modulating its activities and influencing PIC fate.

Purpose of the Study:

  • To investigate the real-time binding dynamics of TFIIE subunits (TFIIEα and TFIIEβ) within human Pol II PICs.
  • To elucidate the role of TFIIH in regulating TFIIE binding kinetics and subunit behavior.
  • To examine the impact of disease-related mutations in TFIIEβ on its stability and kinetic interactions within PICs.

Main Methods:

  • Utilized a purified reconstituted human Pol II transcription system.
  • Employed single-molecule total internal reflection fluorescence microscopy for real-time monitoring.
  • Analyzed TFIIE subunit binding dynamics under varying conditions, including TFIIH presence/absence and disease-related mutations.

Main Results:

  • Observed dynamic, rapid on/off binding of TFIIEα and TFIIEβ as a heterodimer within PICs.
  • TFIIH exclusion led to increased association/dissociation rates and asynchronous behavior of TFIIEα and TFIIEβ.
  • Disease-related TFIIEβ mutations destabilized the subunit and altered its kinetic behavior in PICs.

Conclusions:

  • PICs are dynamic assemblies, not static structures, with GTFs exhibiting complex kinetic behaviors.
  • TFIIH plays a critical role in coordinating the kinetic interactions of TFIIE subunits within PICs.
  • The study provides insights into the structural and kinetic interplay of GTFs in transcription regulation and disease mechanisms.