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Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution
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Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution

Published on: January 16, 2017

The mRNA export protein DBP5 binds RNA and the cytoplasmic nucleoporin NUP214 in a mutually exclusive manner

Holger von Moeller1, Claire Basquin, Elena Conti

  • 1Max-Planck-Institute of Biochemistry, Department of Structural Cell Biology, Am Klopferspitz 18, D-82152 Martinsried, Germany.

Nature Structural & Molecular Biology
|February 17, 2009
PubMed

Insights

The DEAD-box protein DBP5, crucial for mRNA export, is inhibited by NUP214. This interaction, revealed by crystal structures, regulates DBP5

Area of Science:

  • Molecular Biology
  • Structural Biology
  • Cell Biology

Background:

  • DEAD-box helicase DBP5 is vital for messenger RNA (mRNA) export from the nucleus to the cytoplasm in eukaryotic cells.
  • DBP5 functions at the cytoplasmic face of the nuclear pore complex, interacting with RNA and regulatory factors.

Purpose of the Study:

  • To elucidate the structural basis and functional consequences of DBP5 interaction with the nucleoporin NUP214.
  • To understand the mechanism by which NUP214 regulates DBP5 activity during mRNA export.

Main Methods:

  • Determined crystal structures of human DBP5 in complex with RNA and AMPPNP, and with NUP214.
  • Performed in vitro biochemical assays to assess RNA binding and ATPase activities of DBP5.
  • Analyzed conserved residues involved in DBP5-NUP214 interactions.

Main Results:

  • Structural data revealed mutually exclusive binding of DBP5 to nucleic acids and NUP214.
  • NUP214 binding significantly reduced both RNA binding and ATPase activities of DBP5.
  • Interactions involve conserved residues, suggesting a conserved regulatory mechanism.

Conclusions:

  • NUP214 acts as a negative regulator of DBP5, inhibiting its activity through mutually exclusive binding.
  • This provides a mechanistic framework for the regulation of DBP5 during the final stages of mRNA export.
  • Demonstrates a general principle of how regulatory proteins can inhibit DEAD-box family members.

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