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Anaphase Promoting Complex00:50

Anaphase Promoting Complex

The stepwise destruction of specific proteins is necessary for the progression and completion of the cell cycle. Such proteins are ubiquitinated by ubiquitin ligases and then subsequently destroyed by the proteasome. The SCF (Skp1/Cullin/F-box) and the anaphase-promoting complex (APC) are two important ubiquitin ligases involved in cell cycle progression. While SCF is active throughout the cell cycle, APC gets activated during metaphase to anaphase transition. Cdc20 or Cdh1 binds to APC and...
Anaphase Promoting Complex00:50

Anaphase Promoting Complex

The stepwise destruction of specific proteins is necessary for the progression and completion of the cell cycle. Such proteins are ubiquitinated by ubiquitin ligases and then subsequently destroyed by the proteasome. The SCF (Skp1/Cullin/F-box) and the anaphase-promoting complex (APC) are two important ubiquitin ligases involved in cell cycle progression. While SCF is active throughout the cell cycle, APC gets activated during metaphase to anaphase transition. Cdc20 or Cdh1 binds to APC and...
The Spindle Assembly Checkpoint02:19

The Spindle Assembly Checkpoint

The spindle assembly checkpoint is a molecular surveillance mechanism ensuring the fidelity of chromosome segregation during anaphase. The checkpoint monitors the completion of all the prerequisite steps before chromosome segregation to determine whether the segregation process should proceed or be delayed.
Many proteins function together to control the spindle assembly checkpoint. Mutations affecting these proteins may allow cells to proceed into anaphase prematurely, resulting in the...
Transcription Initiation01:47

Transcription Initiation

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...
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

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, a...
SNAREs and Membrane Fusion01:43

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...

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Related Experiment Video

Updated: Jul 10, 2026

Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale
10:50

Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale

Published on: March 14, 2019

A poised initiation complex is activated by SNF1.

Christine Tachibana1, Rhiannon Biddick, G Lynn Law

  • 1Department of Biochemistry, University of Washington, Seattle, WA 98195, USA.

The Journal of Biological Chemistry
|November 3, 2007
PubMed
Summary

AMP kinase (Snf1) is crucial for activating glucose-repressed genes in yeast, working after RNA polymerase II recruitment. Combining Snf1 activation with histone deacetylase mutations and Adr1 enhances gene expression.

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Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale
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Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • Gene Regulation

Background:

  • Snf1, the yeast AMP-activated protein kinase homolog, is vital for derepressing glucose-repressed genes.
  • These genes are activated by transcription factors like Adr1 and Cat8.
  • The exact function of Snf1 in this process, particularly its role in Adr1 DNA binding, remains unclear.

Purpose of the Study:

  • To elucidate the precise role of Snf1 in the activation of glucose-repressed genes by Adr1.
  • To investigate the molecular events occurring after RNA polymerase II recruitment during glucose repression.

Main Methods:

  • Utilized yeast genetics, including deletion of histone deacetylase genes.
  • Analyzed Adr1 and Cat8 DNA binding and recruitment of the preinitiation complex.
  • Assessed RNA polymerase II recruitment and transcriptional activity under various conditions.

Main Results:

  • Deletion of histone deacetylase genes enabled constitutive promoter binding of Adr1 and Cat8.
  • In repressed conditions, Adr1 and Cat8 recruited a partial preinitiation complex including RNA polymerase II to the ADH2 promoter.
  • Transcription initiation was dependent on Snf1 activation, indicating a post-recruitment event.
  • Complete relief of glucose repression was achieved by combining histone deacetylase mutations, Snf1 activation, and a specific Adr1 mutant.

Conclusions:

  • Snf1 plays a critical role in a step following RNA polymerase II recruitment, essential for initiating transcription of glucose-repressed genes.
  • Chromatin remodeling, Snf1 activity, and specific activator function are key components in overcoming glucose repression.