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

PI3K/mTOR/AKT Signaling Pathway01:22

PI3K/mTOR/AKT Signaling Pathway

4.1K
The mammalian target of rapamycin  (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1  (mTORC1) and mTOR complex 2  (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast,  mTORC2 consists of a...
4.1K
TGF - β Signaling Pathway01:16

TGF - β Signaling Pathway

7.9K
The TGF-β signaling pathway regulates cell growth, differentiation, adhesion, motility, and development. TGF-β ligands that induce TGF-β signaling are synthesized in their latent form. Several proteases or cell surface receptors such as integrins act upon the latent form, releasing the active ligand. There are three types of mammalian TGF-βs: (TGF-β1, TGF-β2, and TGF-β3) that bind as homodimers or heterodimers to TGF-β receptors. The TGF-β receptors...
7.9K
MAPK Signaling Cascades01:07

MAPK Signaling Cascades

6.4K
Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
6.4K
The JAK-STAT Signaling Pathway01:20

The JAK-STAT Signaling Pathway

9.5K
Several cytokine receptors have tightly bound Janus kinase or JAK proteins attached at their cytosolic tail. Small signaling molecules such as cytokines, growth hormones, or prolactins bind to the cytokine receptors and initiate their dimerization. The dimerization brings the cytosolic JAKs together that trans-phosphorylate and activates each other. The activated JAKs now phosphorylate cytosolic tails of the cytokine receptors, which serve as binding sites for adaptor proteins such as  SH2...
9.5K
Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

12.3K
When a ligand binds to a cell-surface receptor, the receptor's intracellular domain changes shape, which may either activate its enzyme function or allow its binding to other molecules. The initial signal is amplified by most signal transduction pathways. This means that a single ligand molecule can activate multiple molecules of a downstream target. Proteins that relay a signal are most commonly phosphorylated at one or more sites, activating or inactivating the protein. Kinases catalyze...
12.3K
cAMP-dependent Protein Kinase Pathways01:25

cAMP-dependent Protein Kinase Pathways

6.9K
Cyclic Adenosine Monophosphate (cAMP) is an essential second messenger that activates protein kinase A (PKA) and regulates various biological processes. A single epinephrine molecule binds to GPCR and activates several heterotrimeric G proteins, each stimulating multiple adenylyl cyclase, amplifying the signal, and synthesizing large numbers of cAMP molecules. Small changes in cAMP concentration affect PKA activity. The binding of four cAMP molecules induces a conformational change in PKA,...
6.9K

You might also read

Related Articles

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

Sort by
Same author

Metal-center electron affinity modulates multicolor electrochromism in 2D conjugated metal-organic frameworks.

Nature communications·2026
Same author

Low Tumor miR-369-3p Levels Combined with Plasma Hepatitis B Virus Pre-S2 Gene Deletion Mutation Predict Higher Hepatocellular Carcinoma Recurrence.

Journal of hepatocellular carcinoma·2026
Same author

Mitigation of Ischemia-Reperfusion Injury and Improvement in Overall Graft Viability by Hypothermic Pulsatile Perfusion with Molecular Hydrogen Is Associated with Trx-1/HO-1 Activation in a Non-Survival Ex Vivo Swine Model of Donation-After-Circulatory-Death Kidney Preservation and Transplantation.

International journal of molecular sciences·2026
Same author

Correlating Infrared Surface Thermometry With Core Temperature Assessments in Simulated Cooling and Rewarming of Porcine Kidneys.

Transplantation proceedings·2026
Same author

First Large Comprehensive Core-Laboratory Evaluation of Implantation Depth and Clinical Outcomes in TAVR: Final Global Results from the Optimize PRO Prospective Study.

JACC. Cardiovascular interventions·2026
Same author

Indium-Mediated Glue-Like Interlayer Enables Stable High-Capacity Flexible Sodium Metal Batteries.

Advanced materials (Deerfield Beach, Fla.)·2026

Related Experiment Video

Updated: Oct 5, 2025

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
12:26

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

Published on: May 3, 2018

18.9K

AKT1 mediates multiple phosphorylation events that functionally promote HSF1 activation.

Wen-Cheng Lu1, Ramsey Omari1, Haimanti Ray1

  • 1Medical Sciences, Indiana University School of Medicine, Bloomington, IN, USA.

The FEBS Journal
|January 26, 2022
PubMed
Summary

Heat shock factor 1 (HSF1) activity is regulated by phosphorylation. AKT1 is the most potent kinase, phosphorylating HSF1 at multiple sites including T142, which is crucial for trimerization and gene activation.

Keywords:
AKT1HSF1heat shockphosphorylation

More Related Videos

Oligopeptide Competition Assay for Phosphorylation Site Determination
09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

8.6K
A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF3
11:44

A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF3

Published on: January 24, 2016

12.1K

Related Experiment Videos

Last Updated: Oct 5, 2025

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
12:26

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

Published on: May 3, 2018

18.9K
Oligopeptide Competition Assay for Phosphorylation Site Determination
09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

8.6K
A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF3
11:44

A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF3

Published on: January 24, 2016

12.1K

Area of Science:

  • Molecular Biology
  • Cellular Stress Response

Background:

  • The heat stress response involves the transcription factor heat shock factor 1 (HSF1), which upregulates heat shock proteins to maintain proteome integrity.
  • HSF1 activation is a complex process requiring nuclear localization, trimerization, DNA binding, phosphorylation, and gene transactivation.

Purpose of the Study:

  • To investigate the role of specific kinases, particularly AKT1, in regulating HSF1 activity through phosphorylation.
  • To identify novel phosphorylation sites on HSF1 and elucidate their functional significance in transcriptional regulation.

Main Methods:

  • Kinase assays to determine the ability of AKT1, AKT2, mTOR, p38, MEK1, and DYRK2 to phosphorylate HSF1 at S326.
  • Mass spectrometry to identify all phosphorylation sites on HSF1 following AKT1 treatment.
  • Functional assays to assess the impact of specific phosphorylation sites on HSF1 trimerization, gene transactivation, and TFIIB/CDK9 recruitment.

Main Results:

  • AKT1 was identified as the most potent activator of HSF1 transcriptional activity through S326 phosphorylation, independent of mTOR.
  • Mass spectrometry revealed that AKT1 phosphorylates HSF1 at T142, S230, S326, and T527.
  • Phosphorylation at T142 is essential for HSF1 trimerization, while S230, S326, and T527 are critical for gene transactivation and the recruitment of TFIIB and CDK9.

Conclusions:

  • HSF1 hyperphosphorylation is a targeted process, with specific residues playing direct roles in regulating its transcriptional activity.
  • The novel phosphorylation site T527 in the transactivation domain highlights a new regulatory mechanism for HSF1.
  • AKT1 plays a central role in HSF1 activation through multi-site phosphorylation, impacting key steps in the heat stress response.