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

6.0K
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...
6.0K
The JAK-STAT Signaling Pathway01:20

The JAK-STAT Signaling Pathway

13.5K
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...
13.5K
IP3/DAG Signaling Pathway01:11

IP3/DAG Signaling Pathway

15.4K
Membrane lipids such as phosphatidylinositol (PI) are precursors for several membrane-bound and soluble second messengers. Specific kinases phosphorylate PI and produce phosphorylated inositol phospholipids. One such inositol phospholipids are the  phosphatidylinositol-4,5 bisphosphate [PI(4,5)P2], present in the inner half of the lipid bilayer. Upon ligand binding, GPCR stimulates Gq proteins to turn on phospholipase Cꞵ. Activated phospholipase Cꞵ cleaves PI(4,5)P2 and...
15.4K
Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

18.9K
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...
18.9K
Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

15.4K
Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
15.4K
Regulation of the Unfolded Protein Response01:31

Regulation of the Unfolded Protein Response

3.1K
Inositol-requiring kinase one or IRE1 is the most conserved eukaryotic unfolded protein response (UPR) receptor. It is a type I transmembrane protein kinase receptor with a distinctive site-specific RNase activity. As the binding mechanics of the misfolded proteins with the N-terminal domain of IRE-1 are unclear, three binding models — direct, indirect, and allosteric -- are proposed for receptor activation. Nevertheless, it is known that once a misfolded protein associates with IRE1, it...
3.1K

You might also read

Related Articles

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

Sort by
Same author

The ISG Atlas: a loss-of-function analysis characterizes antiviral properties of interferon stimulated genes.

Nature communications·2026
Same author

HCR-Proxy resolves site-specific proximal RNA microenvironments at subcompartmental resolution.

Nucleic acids research·2026
Same author

Correction: RBM39 shapes innate immunity by controlling expression of key factors of the interferon response.

Frontiers in immunology·2026
Same author

Modanovo: A Unified Model for Post-translational Modification-Aware De Novo Sequencing Using Experimental Spectra From In Vivo and Synthetic Peptides.

Molecular & cellular proteomics : MCP·2025
Same author

Elongator is required for pattern recognition receptor and type I interferon signaling in macrophages.

The Journal of biological chemistry·2025
Same author

Host cell Z-RNAs activate ZBP1 during virus infections.

Nature·2025

Related Experiment Video

Updated: Mar 8, 2026

Author Spotlight: Developing Tools to Tune the Activity of Tyrosine Phosphatases
06:56

Author Spotlight: Developing Tools to Tune the Activity of Tyrosine Phosphatases

Published on: September 6, 2024

847

Phosphorylation-Dependent Feedback Inhibition of RIG-I by DAPK1 Identified by Kinome-wide siRNA Screening.

Joschka Willemsen1, Oliver Wicht2, Julia C Wolanski3

  • 1Research Group "Dynamics of early viral infection and the innate antiviral response," Division Virus-associated carcinogenesis (F170), German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany; Department for Infectious Diseases, Molecular Virology, Research Group "Dynamics of early viral infection and the innate antiviral response," Medical Faculty, Heidelberg University, 69120 Heidelberg, Germany.

Molecular Cell
|January 31, 2017
PubMed
Summary

A negative-feedback loop involving death associated protein kinase 1 (DAPK1) regulates type I interferon production. DAPK1 activation inhibits RIG-I signaling, preventing excessive immune responses and potential autoimmune disease.

Keywords:
DAPK1DDX58RIG-Iantiviral responsecytokinesfeedback regulationinnate immunityinterferon systempattern recognition receptorssignal transduction

More Related Videos

Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag
08:55

Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag

Published on: December 14, 2017

16.2K
Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

8.6K

Related Experiment Videos

Last Updated: Mar 8, 2026

Author Spotlight: Developing Tools to Tune the Activity of Tyrosine Phosphatases
06:56

Author Spotlight: Developing Tools to Tune the Activity of Tyrosine Phosphatases

Published on: September 6, 2024

847
Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag
08:55

Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag

Published on: December 14, 2017

16.2K
Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

8.6K

Area of Science:

  • Immunology
  • Molecular Biology
  • Virology

Background:

  • Type I interferon induction requires stringent regulation for effective antiviral defense and prevention of immunopathology.
  • Dysregulated interferon signaling is implicated in autoimmune diseases.
  • RIG-I (retinoic acid-inducible gene I) is a key sensor of viral RNA that initiates interferon production.

Purpose of the Study:

  • To identify novel regulators of RIG-I/IRF3 signaling.
  • To elucidate the mechanisms controlling type I interferon induction and limitation.
  • To characterize a newly identified negative-feedback pathway in interferon regulation.

Main Methods:

  • Genome-wide RNA interference (RNAi) screening of kinases.
  • Validation of identified regulatory factors.
  • In vitro kinase assays and phosphorylation site analysis.

Main Results:

  • Identified 22 factors regulating RIG-I/IRF3 signaling.
  • Discovered a negative-feedback mechanism mediated by death associated protein kinase 1 (DAPK1).
  • DAPK1 activation by RIG-I signaling inhibits IRF3 activity and subsequent interferon-beta production.
  • DAPK1 phosphorylates RIG-I, limiting its ability to sense viral RNA and activate signaling.

Conclusions:

  • DAPK1 acts as a crucial negative regulator of RIG-I signaling.
  • This DAPK1-mediated feedback loop is essential for controlling interferon responses.
  • Proper regulation of RIG-I by DAPK1 prevents excessive immune activation and potential autoimmunity.