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Nucleocytoplasmic transport of luciferase gene mRNA requires CRM1/Exportin1 and RanGTPase

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Abstract

Human immunodeficiency virus type 1 Rev (regulator of the expression of the virion) protein was shown to reduce the expression level of the co-transfected luciferase reporter gene (luc+) introduced to monitor transfection efficiency. We studied the mechanism of the inhibitory Rev effect. The effect, caused by nuclear retention of luc+ mRNA, was reversed if rev had a point mutation that makes its nuclear export signal (NES) unable to associate with cellular transport factors. The Rev NES receptor CRM1 (chromosome region maintenance 1)-specific inhibitor, leptomycin B, blocked luc+ mRNA export. This finding was also supported by the overexpression of ΔCAN, another specific CRM1 inhibitor that caused inhibition of luciferase gene expression. Experiments involving tsBN2 cells, which have a temperature-sensitive RCC1 (regulator of chromosome condensation 1) allele, demonstrated that luc+ expression required generation of the GTP-bound form of RanGTPase (RanGTP) by RCC1. The constitutive transport element (CTE)-mediated nuclear export of luc+ mRNA was found to also depend upon RanGTP. Nuclear export of luc+ mRNA is thus suggested to involve CRM1 and RanGTP, which Rev employs to transport viral mRNA. The Rev effect is therefore considered to involve competition between two molecules for common transport factors.

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Authors and Affiliations

  1. Laboratory of Microbiology and Cell Biology, Department of Pharmacy, College of Pharmaceutical Sciences, Ritsumeikan University, Kusatsu, Shiga, 525-8577, Japan

    Tominori Kimura & Iwao Hashimoto

  2. Department of Microbiology, Kansai Medical University, Osaka, Japan

    Tominori Kimura, Iwao Hashimoto & Masao Nishikawa

  3. Department of Anatomy and Cell Science, Kansai Medical University, Osaka, Japan

    Hisao Yamada

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  1. Tominori Kimura
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  2. Iwao Hashimoto
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Correspondence to Tominori Kimura.

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Kimura, T., Hashimoto, I., Nishikawa, M. et al. Nucleocytoplasmic transport of luciferase gene mRNA requires CRM1/Exportin1 and RanGTPase. Med Mol Morphol 42, 70–81 (2009). https://doi.org/10.1007/s00795-009-0441-3

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  • DOI: https://doi.org/10.1007/s00795-009-0441-3

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