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MKRN2 Physically Interacts with GLE1 to Regulate mRNA Export and Zebrafish Retinal Development.

Eric J Wolf1, Amanda Miles2, Eliza S Lee3

  • 1Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.

Cell Reports
|May 28, 2020
PubMed
Summary

The RNA-binding E3 ubiquitin ligase MKRN2 interacts with GLE1 to regulate messenger RNA (mRNA) nuclear export and retinal development. This finding reveals a novel mechanism controlling gene expression and cellular processes.

Keywords:
GLE1MKRN2RNA binding proteinsaffinity purification mass spectrometrymRNA exportzebrafish retinogenesis

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Messenger RNA (mRNA) nuclear export is crucial for gene expression, involving complex protein interactions at the nuclear pore complex.
  • Ribonucleoprotein remodeling is a key step in mRNA export termination.
  • The role of RNA-binding E3 ubiquitin ligases in mRNA export is not fully understood.

Purpose of the Study:

  • To identify novel protein interactions involved in mammalian mRNA nuclear export.
  • To investigate the functional association between MKRN2 and GLE1 in mRNA export and retinal development.
  • To elucidate the mechanism by which MKRN2 regulates mRNA export.

Main Methods:

  • Affinity-purification mass spectrometry to screen protein interactions of human RNA-binding E3 ubiquitin ligases.
  • Zebrafish model to assess epistasis between MKRN2 and GLE1.
  • Morpholino-mediated knockdown and CRISPR/Cas9 knockout in zebrafish.
  • Ribonomic approaches to identify mRNA targets of MKRN2.

Main Results:

  • Identified a physical interaction between MKRN2 and GLE1, an mRNA export factor.
  • MKRN2 knockdown or knockout partially rescued retinal defects caused by GLE1 depletion in zebrafish.
  • MKRN2 selectively binds to the 3' UTR of a subset of mRNAs.
  • Nuclear export of MKRN2-associated mRNAs was enhanced upon MKRN2 knockdown.

Conclusions:

  • MKRN2 interacts with GLE1 to selectively regulate mRNA nuclear export.
  • This interaction plays a role in retinal development.
  • MKRN2 acts as a regulator of mRNA export, influencing gene expression and cellular processes.