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RPS28B mRNA acts as a scaffold promoting cis-translational interaction of proteins driving P-body assembly

Nikita Fernandes1, J Ross Buchan1

  • 1Department of Molecular and Cellular Biology, University of Arizona, Tucson, AZ 85721, USA.

Nucleic Acids Research
|May 13, 2020
PubMed

Insights

Specific mRNA sequences, like RPS28B 3'UTR, can nucleate the assembly of cytoplasmic P-bodies (PB). This occurs when the cis-translated Rps28 protein interacts with Edc3, facilitating PB formation.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • P-bodies (PBs) are essential cytoplasmic mRNA-protein (mRNP) granules involved in mRNA regulation.
  • PB assembly is typically driven by self-interacting proteins, but the role of specific mRNAs is less understood.
  • Rps28b mutants fail to form PBs, suggesting a role for this gene in PB assembly.

Purpose of the Study:

  • To investigate the role of specific mRNA transcripts in facilitating PB assembly.
  • To determine if the RPS28B 3'UTR is critical for PB formation.
  • To elucidate the mechanism by which Rps28 influences PB assembly.

Main Methods:

  • Analysis of rps28b mutants and their PB formation under normal conditions.
  • Expression studies of the RPS28B 3'UTR alone.
  • Chimeric mRNA studies to compare cis- and trans-translation effects on protein-RNA interactions.
  • Investigating the interaction between Rps28 protein and Edc3.

Main Results:

  • The RPS28B 3'UTR is important for PB assembly and binds to the PB protein Edc3.
  • Expression of the RPS28B 3'UTR alone does not drive PB assembly.
  • Rps28 protein translated in cis from an mRNA with the RPS28B 3'UTR interacts more strongly with Edc3 than Rps28 translated in trans.
  • This cis-translation-mediated interaction facilitates PB assembly.

Conclusions:

  • Specific mRNA sequences, termed 'RNA scaffolds,' can nucleate PB assembly.
  • A cis-translated protein (Rps28) interacting with a protein in the mRNA's 3'UTR (Edc3) can stimulate cellular structure assembly.
  • This study describes a novel mechanism for PB assembly initiation in yeast involving cis-translated proteins and 3'UTRs.

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