The cellular DExD/H-box RNA-helicases UAP56 and URH49 exhibit a CRM1-independent nucleocytoplasmic shuttling activity

PLoS One. 2011;6(7):e22671. doi: 10.1371/journal.pone.0022671. Epub 2011 Jul 22.

Abstract

Cellular DExD/H-box RNA-helicases perform essential functions during mRNA biogenesis. The closely related human proteins UAP56 and URH49 are members of this protein family and play an essential role for cellular mRNA export by recruiting the adaptor protein REF to spliced and unspliced mRNAs. In order to gain insight into their mode of action, we aimed to characterize these RNA-helicases in more detail. Here, we demonstrate that UAP56 and URH49 exhibit an intrinsic CRM1-independent nucleocytoplasmic shuttling activity. Extensive mapping studies identified distinct regions within UAP56 or URH49 required for (i) intranuclear localization (UAP56 aa81-381) and (ii) interaction with REF (UAP56 aa51-428). Moreover, the region conferring nucleocytoplasmic shuttling activity was mapped to the C-terminus of UAP56, comprising the amino acids 195-428. Interestingly, this region coincides with a domain within Uap56p of S. pombe that has been reported to be required for both Rae1p-interaction and nucleocytoplasmic shuttling. However, in contrast to this finding we report that human UAP56 shuttles independently from Rae1. In summary, our results reveal nucleocytoplasmic shuttling as a conserved feature of yeast and human UAP56, while their export receptor seems to have diverged during evolution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Active Transport, Cell Nucleus
  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Cell Nucleus / metabolism*
  • Cytoplasm / metabolism*
  • DEAD-box RNA Helicases / chemistry
  • DEAD-box RNA Helicases / genetics
  • DEAD-box RNA Helicases / metabolism*
  • Exportin 1 Protein
  • HEK293 Cells
  • Humans
  • Karyopherins / metabolism*
  • Mice
  • Molecular Sequence Data
  • Mutagenesis
  • Mutation
  • NIH 3T3 Cells
  • Nuclear Matrix-Associated Proteins / metabolism
  • Nucleocytoplasmic Transport Proteins / metabolism
  • Protein Structure, Tertiary
  • Receptors, Cytoplasmic and Nuclear / metabolism*

Substances

  • Karyopherins
  • Nuclear Matrix-Associated Proteins
  • Nucleocytoplasmic Transport Proteins
  • RAE1 protein, human
  • Receptors, Cytoplasmic and Nuclear
  • DDX39A protein, human
  • DDX39B protein, human
  • DEAD-box RNA Helicases