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J. Biol. Chem., Vol. 282, Issue 21, 15645-15651, May 25, 2007

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The Interaction between Cap-binding Complex and RNA Export Factor Is Required for Intronless mRNA Export^*

Takayuki Nojima, Tetsuro Hirose^¶||, Hiroshi Kimura^**, and Masatoshi Hagiwara¹

From the Laboratory of Gene Expression, School of Biomedical Science, the Department of Functional Genomics, Medical Research Institute, Tokyo Medical and Dental University, Yushima 1-5-45, Bunkyo-ku, Tokyo 113-8510, the ^¶Biological Information Research Center, National Institute of Advanced Industrial Science and Technology (AIST), Aomi 2-42, Koto-Ku, Tokyo 135-0064, ^||PRESTO, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama, and the ^**Nuclear Function and Dynamics Unit, HMRO, Graduate School of Medicine, Kyoto University, Yoshida Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan

RNA export factor (REF) is a component of the exon junction complex (EJC) that is deposited on mRNA in a splicing-dependent manner, and targets spliced mRNA for export. In this study, analysis of the RNA-binding protein complexes revealed that REF associates with -globin mRNA at the region other than the EJC deposition site. Comparison between RNA polymerase II and T7 transcription and further analysis showed that the deposition of REF apart from the EJC is dependent on the 5' cap structure, but not splicing. Excess amounts of m⁷GpppG cap analog reduced REF binding to intronless mRNA, and a co-immunoprecipitation experiment revealed that REF interacts with the cap-binding protein CBP20. The export of Cy3-labeled intronless -globin mRNA from nuclei of HeLa cells was enhanced by co-injection of CBP20 and REF. Thus, REF recruited by CBP20 may play a stimulatory role to export the capped intronless mRNAs.

Received for publication, January 23, 2007 , and in revised form, March 15, 2007.

^* This work was supported by grants-in-aid (to M. H.) from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan, the National Institute of Biomedical Innovation (NIBI), and the 21st COE Program for Frontier Research on Molecular Destruction and Reconstruction of Tooth and Bone, represented by Masaki Noda. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. S1 and S2.

¹ To whom correspondence should be addressed. Tel.: 81-3-5803-5836; Fax: 81-3-5803-5853; E-mail: m.hagiwara.end{at}mri.tmd.ac.jp.

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