Related Experiment Video

Updated: Jul 11, 2025

Identification of Transcription Factor Regulators using Medium-Throughput Screening of Arrayed Libraries and a Dual-Luciferase-Based Reporter

Identification of Transcription Factor Regulators using Medium-Throughput Screening of Arrayed Libraries and a Dual-Luciferase-Based Reporter

Published on: March 27, 2020

Neuronal survival factor TAFA2 suppresses apoptosis through binding to ADGRL1 and activating cAMP/PKA/CREB/BCL2

Hui Liang¹, Ling Yun Tang¹, Hao Yang Ge¹

¹State Key Laboratory of Medical Genomics, Research Center for Experimental Medicine, Rui-Jin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.

|November 9, 2023

Abstract

Aims:

TAFA2, a cytokine specifically expressed in the central nervous system, plays a vital role in neuronal cell survival. TAFA2 deficiency has been correlated to various neurological disorders in mice and humans. However, the underlying mechanism remains elusive, especially its membrane-binding receptor through which TAFA2 functions. This study aimed to identify the specific binding receptor responsible for the anti-apoptotic effects of TAFA2.

Main Method:

Co-immunoprecipitation (Co-IP) and quantitative mass spectrometry-based proteomic analysis were employed to identify potential TAFA2 binding proteins in V5 knockin mouse brain lysates. Subsequent validation involved in vitro and in vivo Co-IP and pull-down using specific antibodies. The functional analysis included evaluating the effects of ADGRL1 knockout, overexpression, and Lectin-like domain (Lec) deletion mutant on TAFA2's anti-apoptotic activity and analyzing the intracellular signaling pathways mediated by TAFA2 through ADGRL1.

Key Findings:

Our study identified ADGRL1 as a potential receptor for TAFA2, which directly binds to TAFA2 through its lectin-like domain. Overexpression ADGRL1, but not ADGRL1Δ^Lec, induced apoptosis, which could be effectively suppressed by recombinant TAFA2 (rTAFA2). In ADGRL1^-/- cells or re-introducing with ADGRL1Δ^Lec, responses to rTAFA2 in suppressing cell apoptosis were compromised. Increased cAMP, p-PKA, p-CREB, and BCL2 levels were also observed in response to rTAFA2 treatment, with these responses attenuated in ADGRL1^-/- or ADGRL1Δ^Lec-expressing cells.

Significance:

Our results demonstrated that TAFA2 directly binds to the lectin-like domain of ADGRL1, activating cAMP/PKA/CREB/BCL2 signaling pathway, which is crucial in preventing cell death. These results implicate TAFA2 and its receptor ADGRL1 as potential therapeutic targets for neurological disorders.

Insights

Researchers identified ADGRL1 as the receptor for TAFA2, a central nervous system cytokine. This interaction activates a signaling pathway crucial for neuronal survival and preventing cell death, offering potential therapeutic targets for neurological disorders.

Area of Science:

Neuroscience
Cell Biology
Molecular Biology

Background:

TAFA2 is a central nervous system cytokine vital for neuronal survival.
TAFA2 deficiency is linked to neurological disorders, but its receptor and mechanism are unknown.
Identifying the TAFA2 receptor is crucial for understanding its anti-apoptotic function.

Purpose of the Study:

To identify the specific membrane-binding receptor for TAFA2.
To elucidate the mechanism underlying TAFA2's anti-apoptotic effects.
To investigate the role of ADGRL1 in TAFA2-mediated neuronal survival.

Main Methods:

Co-immunoprecipitation and mass spectrometry to identify TAFA2 binding proteins.
In vitro and in vivo validation of TAFA2-ADGRL1 interaction.
Functional assays using ADGRL1 knockout/overexpression and mutant analysis.
Analysis of intracellular signaling pathways (cAMP, PKA, CREB, BCL2).

Main Results:

ADGRL1 was identified as a direct binding receptor for TAFA2 via its lectin-like domain.
ADGRL1 overexpression induced apoptosis, suppressed by TAFA2; ADGRL1 deficiency or mutant expression compromised TAFA2's protective effect.
TAFA2 treatment increased cAMP, p-PKA, p-CREB, and BCL2 levels, dependent on ADGRL1 and its lectin-like domain.

Conclusions:

TAFA2 directly binds the lectin-like domain of ADGRL1 to prevent cell death.
The cAMP/PKA/CREB/BCL2 signaling pathway mediates TAFA2's anti-apoptotic function through ADGRL1.
TAFA2 and ADGRL1 represent potential therapeutic targets for neurological diseases.

Read full analysis

Keywords:

FAM19A2 GPCR Knock-in mice Lectin-like domain Ligand-receptor interaction

More Related Videos

Protocol for Culturing Sympathetic Neurons from Rat Superior Cervical Ganglia SCG

Protocol for Culturing Sympathetic Neurons from Rat Superior Cervical Ganglia SCG

Published on: January 30, 2009

Identification of Intracellular Signaling Events Induced in Viable Cells by Interaction with Neighboring Cells Undergoing Apoptotic Cell Death

Identification of Intracellular Signaling Events Induced in Viable Cells by Interaction with Neighboring Cells Undergoing Apoptotic Cell Death

Published on: December 27, 2016

Related Concept Videos

MAPK Signaling Cascades

MAPK Signaling Cascades

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...

The Extrinsic Apoptotic Pathway

The Extrinsic Apoptotic Pathway

The extrinsic apoptotic pathway is initiated when extracellular death-inducing signals, such as specific cytokines, activate the death receptors expressed on the cell surface. The immune cells involved in this pathway are natural killer cells (NK cells) and cytotoxic T-lymphocytes. NK cells are critical in innate immune response, while cytotoxic T-lymphocytes are associated with adaptive immune response. These cells recognize specific receptors expressed on the altered cells and activate...

The Intrinsic Apoptotic Pathway

The Intrinsic Apoptotic Pathway

Internal cellular stress, such as cellular injury or hypoxia, triggers intrinsic apoptosis. The B-cell lymphoma 2 (Bcl-2) family of proteins are the primary regulators of the intrinsic apoptotic pathway. For example, during DNA damage, checkpoint proteins, such as Ataxia Telangiectasia Mutated (ATM protein) and Checkpoints Factor-2 (Chk2) proteins, are activated. These proteins phosphorylate p53 which further activates pro-apoptotic proteins, such as Bax, Bak, PUMA, and Noxa, and inhibits...

PI3K/mTOR/AKT Signaling Pathway

PI3K/mTOR/AKT Signaling Pathway

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...

Enzyme-linked Receptors

Enzyme-linked Receptors

Enzyme-linked receptors are proteins that act as both receptor and enzyme, activating multiple intracellular signals. This is a large group of receptors that include the receptor tyrosine kinase (RTK) family. Many growth factors and hormones bind to and activate the RTKs.
Neurotrophin (NT) receptors are a family of RTKs, including trkA, trkB, and trkC (tropomyosin-related kinase) receptors. TrkA is specific for nerve growth factor (NGF), neurotrophin-6, and neurotrophin-7. TrkB binds...

TGF - β Signaling Pathway

TGF - β Signaling Pathway

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...

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

Partially charged platinum on aminated and carboxylated SBA-15 as a catalyst for alkene hydrosilylation.

RSC advances·2022

Peroxisome proliferator-activated receptor-γ ameliorates neuronal ferroptosis after traumatic brain injury in mice by inhibiting cyclooxygenase-2.

Experimental neurology·2022

Catalytic wet peroxide degradation of acrylonitrile wastewater by ordered mesoporous Ag/CeO2: synthesis, performance and kinetics.

RSC advances·2022

Molecular chaperone heat shock protein 70 inhibitors suppress conditioned place preference induced by morphine exposure in male rats.

Addiction biology·2022

Case Report and Literature Review: Bacterial Meningoencephalitis or Not? Naegleria fowleri Related Primary Amoebic Meningoencephalitis in China.

Frontiers in pediatrics·2022

Folic acid intervention changes liver Foxp3 methylation and ameliorates the damage caused by Th17/Treg imbalance after long-term alcohol exposure.

Food & function·2022

See all related articles