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Medical Molecular Morphology

, Volume 43, Issue 3, pp 145–157 | Cite as

Novel cis-active structures in the coding region mediate CRM1-dependent nuclear export of IFN-α 1 mRNA

  • Tominori KimuraEmail author
  • Iwao Hashimoto
  • Mikio Nishizawa
  • Seiji Ito
  • Hisao Yamada
Original Paper

Abstract

We recently reported the chromosome region maintenance 1 (CRM1)-dependent nuclear export of intron-less human interferon-α1 (IFN-α1) mRNA, which encodes a main effecter of host innate immunity. We show that the coding region of IFN-α1 mRNA forms novel secondary structures that are responsible for the CRM1-dependent export of the transcript. Deletion-mutagenesis, in vivo export assays, and computer analyses of the folding potentials of export-competent fragments revealed the presence of a domain, termed the conserved secondary structure (CSS), comprising two adjacent putative stable stem-loop structures (nt 208–452). Internal deletion-mutagenesis and constitutive export assays of each stem-loop structure demonstrated that subregions 308–322 and 352–434 act as a core element by conferring the export function on the CSS. Leptomycin B (LMB) inhibition of the CRM1 pathway decreased the export of core element RNA, implying that the principal site of CRM1 action for exporting IFN-α1 mRNA resides within the core element. An RNPS1 (RNA-binding protein S1, serine-rich domain) cDNA was isolated by yeast three-hybrid screening, using bait containing two CSS regions. We showed that RNPS1 might recognize IFN-α1 mRNP that includes CRM1. The data demonstrate that interaction between RNA structures in the coding region and CRM1 affects the nucleocytoplasmic translocation of IFN-α1 mRNA.

Key words

Nuclear export IFN-α1 mRNA RNA secondary structure CRM1 RNA-fluorescence in situ hybridization Yeast three-hybrid assay RNPS1 

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Copyright information

© The Japanese Society for Clinical Molecular Morphology 2010

Authors and Affiliations

  • Tominori Kimura
    • 1
    • 3
    Email author
  • Iwao Hashimoto
    • 1
    • 3
  • Mikio Nishizawa
    • 2
    • 4
  • Seiji Ito
    • 4
  • Hisao Yamada
    • 5
  1. 1.Laboratory of Microbiology and Cell Biology, Department of Pharmacy, College of Pharmaceutical SciencesRitsumeikan UniversityKusatsu, ShigaJapan
  2. 2.Laboratory of Medical Chemistry, Department of Biomedical Sciences, College of Life SciencesRitsumeikan UniversityShigaJapan
  3. 3.Department of MicrobiologyKansai Medical UniversityOsakaJapan
  4. 4.Department of Medical ChemistryKansai Medical UniversityOsakaJapan
  5. 5.Department of Anatomy and Cell ScienceKansai Medical UniversityOsakaJapan

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