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First published online May 4, 2004
doi: 10.1242/10.1242/jcs.01076

Journal of Cell Science 117, 2259-2270 (2004)
Published by The Company of Biologists 2004
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Research Article

CRM1-dependent, but not ARE-mediated, nuclear export of IFN-{alpha}1 mRNA

Tominori Kimura1,*, Iwao Hashimoto1, Takahiro Nagase2 and Jun-Ichi Fujisawa1,*

1 Department of Microbiology, Kansai Medical University, Moriguchi, Osaka 570-8506, Japan
2 Kazusa DNA Research Institute, Kazusa-kamatari, Chiba 292-0818, Japan

* Authors for correspondence (e-mail: fujisawa{at}takii.kmu.ac.jp; kimura{at}takii.kmu.ac.jp)

Accepted 5 January 2004

While the bulk of cellular mRNA is known to be exported by the TAP pathway, export of specific subsets of cellular mRNAs may rely on chromosome region maintenance 1 (CRM1). One line of evidence supporting this hypothesis comes from the study of mRNAs of certain early response genes (ERGs) containing the adenylate uridylate-rich element (ARE) in their 3' untranslated regions (3' UTRs). It was reported that HuR-mediated nuclear export of these mRNAs was CRM1-dependent under certain stress conditions. To further examine potential CRM1 pathways for other cellular mRNAs under stress conditions, the nuclear export of human interferon-{alpha}1 (IFN-{alpha}1) mRNA, an ERG mRNA induced upon viral infection, was studied. Overproduction of human immunodeficiency virus type 1 Rev protein reduced the expression level of the co-transfected IFN-{alpha}1 gene. This inhibitory effect, resulting from nuclear retention of IFN-{alpha}1 mRNA, was reversed when rev had a point mutation that made its nuclear export signal unable to associate with CRM1. Leptomycin B sensitivity experiments revealed that the cytoplasmic expression of IFN-{alpha}1 mRNA was arrested upon inhibition of CRM1. This finding was further supported by overexpression of {Delta}CAN, a defective form of the nucleoporin Nup214/CAN that inhibits CRM1 in a dominant-negative manner, which resulted in the effective inhibition of IFN-{alpha}1 gene expression. Subsequent RNA fluorescence in situ hybridisation and immunocytochemistry demonstrated that the IFN-{alpha}1 mRNA was colocalised with CRM1, but not with TAP, in the nucleus. These results therefore imply that the nuclear export of IFN-{alpha}1 mRNA is mediated by CRM1. However, truncation of the 3' UTR did not negatively affect the nuclear export of IFN-{alpha}1 mRNA that lacked the ARE, unexpectedly indicating that this CRM1-dependent mRNA export may not be mediated via the ARE.

Key words: Nuclear export, interferon-{alpha}1 mRNA, CRM1, HIV-1 Rev


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CRM1 and IFN-{alpha}1 mRNA: nuclear export partners

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