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Heterotrimeric G proteins are guanine nucleotide-binding proteins. As the name suggests, heterotrimeric G proteins are composed of three subunits: alpha, beta, and gamma. They remain GDP-bound or GTP-bound inside the cells and switch between inactive/active states. The Gα subunit possesses the nucleotide-binding pocket that binds guanine nucleotides and switches between GDP or GTP-bound states. In contrast, the Gꞵ and Gγ subunits are always bound together with high...
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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...
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Activation of Gαq sequesters specific transcripts into Ago2 particles.

Lela Jackson1, Madison Rennie1, Alison Poussaint1

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Gαq signaling activates cytosolic phospholipase Cβ1 (PLCβ1), releasing proteins from Argonaute 2 (Ago2) stress granules. This process selectively protects specific mRNA transcripts, revealing a novel Gαq/PLCβ pathway regulating protein translation.

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Area of Science:

  • Cellular signaling pathways
  • Molecular biology
  • Stress granule dynamics

Background:

  • The Gαq/phospholipase Cβ1 (PLCβ1) system mediates cellular calcium responses.
  • Cytosolic PLCβ1 interacts with Argonaute 2 (Ago2) and proteins in stress granules, inhibiting their aggregation.
  • Gαq activation triggers the relocalization of cytosolic PLCβ1 to the plasma membrane, releasing proteins and promoting stress granule formation.

Purpose of the Study:

  • To characterize Argonaute 2 (Ago2) stress granules associated with Gαq activation.
  • To investigate the impact of Gαq signaling on stress granule composition and RNA content.
  • To identify specific transcripts regulated by the Gαq/PLCβ pathway.

Main Methods:

  • Cellular stimulation with Gαq agonists, heat shock, and osmotic stress in differentiated PC12 cells.
  • Characterization of Ago2-associated stress granules using proteomic and transcriptomic analyses.
  • Reverse transcription polymerase chain reaction (RT-PCR) and western blotting to validate findings.

Main Results:

  • Ago2 stress granules exhibit altered protein composition upon stimulation with Gαq agonists or various stresses.
  • Purified Ago2 granules from stressed cells contain diverse mRNAs and microRNAs (miRs), while control granules lack RNA.
  • Gαq-stimulated Ago2 particles uniquely contain chromogranin B and ATP synthase 5f1b transcripts, suggesting selective protection.

Conclusions:

  • Gαq activation induces unique stress granule formation with specific RNA cargo.
  • The Gαq/PLCβ pathway selectively protects chromogranin B and ATP synthase 5f1b transcripts from degradation.
  • This study reveals a novel mechanism where Gαq/PLCβ signaling regulates the translation of specific proteins.