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Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution
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Dbp5 associates with RNA-bound Mex67 and Nab2 and its localization at the nuclear pore complex is sufficient for mRNP

Rebecca L Adams1, Susan R Wente1

  • 1Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America.

Plos Genetics
|October 1, 2020
PubMed

Insights

The DEAD-box protein Dbp5 is essential for releasing mRNA export factors Mex67 and Nab2 at the nuclear pore complex (NPC). Localizing Dbp5 to the NPC is sufficient for viability, suggesting its mRNA remodeling functions are separable.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Mex67 acts as an mRNA export receptor, interacting with adaptors like Nab2 to facilitate transport through the nuclear pore complex (NPC).
  • Dbp5, a DEAD-box protein, mediates mRNP remodeling at the NPC's cytoplasmic face, releasing export factors during mRNA export.
  • The precise mechanism of selective protein release during Dbp5-mediated remodeling remains unclear.

Purpose of the Study:

  • To investigate the mechanism by which Dbp5 selectively releases proteins during mRNA export.
  • To determine if Dbp5's localization to the NPC is critical for its mRNP remodeling function.
  • To explore the relationship between Dbp5's various cellular functions in mRNA processing.

Main Methods:

  • Co-immunoprecipitation to detect protein complex formation between Dbp5, Mex67, and Nab2.
  • Genetic analysis involving fusion of Dbp5 to Nup159 to anchor it at the NPC.
  • Cell viability assays to assess the functional consequences of Dbp5 localization.

Main Results:

  • Dbp5 was found in close proximity with Mex67 and Nab2 within a cellular complex.
  • Anchoring Dbp5 to the cytoplasmic face of the NPC via fusion to Nup159 supported cell viability.
  • These findings indicate that NPC localization is crucial for Dbp5's essential role in mRNP remodeling.

Conclusions:

  • Dbp5's essential function in remodeling exporting mRNPs relies on its NPC localization.
  • Dbp5's role in mRNA export is separable from its other subcellular functions.
  • This supports a model where Dbp5 is locally recruited to specific transcript regions for targeted removal of transport proteins at the NPC.

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