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Visualization of Endoplasmic Reticulum Localized mRNAs in Mammalian Cells
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Cell-cycle-dependent mRNA localization in P-bodies.

Adham Safieddine1, Marie-Noëlle Benassy1, Thomas Bonte2

  • 1Sorbonne Université, CNRS, Institut de Biologie Paris Seine (IBPS), Laboratoire de Biologie du Développement, 75005 Paris, France.

Molecular Cell
|October 5, 2024
PubMed
Summary
This summary is machine-generated.

P-bodies (PBs) dynamically change their RNA content throughout the cell cycle, revealing complex regulation beyond simple storage of untranslated mRNAs. This research uncovers timely mRNA capture and release mechanisms.

Keywords:
P-bodiesRNA imagingRNA localizationcell cyclemembraneless organelles

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • RNA Biology

Background:

  • Membraneless organelles, such as P-bodies (PBs), play crucial roles in RNA regulation.
  • Understanding the dynamic nature of RNA targeting to PBs is key to elucidating their functions.
  • The cell cycle influences various cellular processes, including RNA metabolism.

Purpose of the Study:

  • To investigate the dynamic changes in P-bodies (PBs) and their RNA content during the cell cycle in HEK293 cells.
  • To determine how mRNA localization to PBs correlates with cell-cycle progression and protein expression.
  • To explore the regulatory mechanisms controlling cyclic mRNA localization in PBs.

Main Methods:

  • Purification of PBs across different phases of the cell cycle.
  • Analysis of RNA content within purified PBs.
  • Single-molecule fluorescence in situ hybridization (FISH) to visualize mRNA localization patterns.
  • Assessment of RNA features (e.g., AU richness, length) and RNA-binding protein (RBP) involvement.

Main Results:

  • Widespread changes in PB RNA content were observed throughout the cell cycle, partially independent of global RNA expression changes.
  • Distinct mRNA localization patterns in PBs were identified, peaking in G1, S, or G2 phases.
  • Examples showed timely mRNA capture in PBs when the encoded protein was no longer essential.
  • Cyclic mRNA localization is regulated by RBPs (e.g., HuR in G2) and specific RNA features, not solely by translation status.
  • PB-localized mRNAs are AU-rich but are longer in G1, potentially linked to post-mitotic PB reassembly.

Conclusions:

  • P-bodies are dynamic regulatory hubs whose RNA composition significantly changes during the cell cycle.
  • mRNA localization to PBs is a regulated process influenced by cell-cycle phase, RBPs, and RNA characteristics.
  • PBs function beyond a passive reservoir for untranslated mRNAs, actively participating in RNA management throughout the cell cycle.