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Nucleic Acids Research 2005 33(9):2929-2941; doi:10.1093/nar/gki605
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Published online 20 May 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

Article

Internal initiation of translation of the TrkB mRNA is mediated by multiple regions within the 5' leader

Tara Dobson, Angela Minic, Kirsten Nielsen, Elizabeth Amiott and Les Krushel*

Department of Pharmacology, University of Colorado Health Sciences Center Denver, CO 80262, USA

*To whom correspondence should be addressed. Tel: +1 303 724 3646; Fax: +1 303 724 3647; Email: leslie.krushel{at}uchsc.edu

Received February 22, 2005. Revised May 2, 2005. Accepted May 2, 2005.

Translational regulation of the dendritically localized mRNA encoding for the neurotrophin receptor TrkB has important ramifications for synaptic function. We examined whether the TrkB mRNA is translated through an internal initiation entry site (IRES). The human TrkB 5' leaders are derived from the use of alternative promoters and alternative splicing, but all 5' leaders share a common exon. Insertion of a full-length 5' leader, as well as the common exon into the intercistronic region of a dicistronic luciferase construct, yielded luciferase activity generated from the second cistron that was either equivalent or higher than that observed from the encephalomyocarditis virus IRES. Moreover, inhibiting cap-dependent translation ex vivo and in in vitro lysates had only a minimal effect on the translation of mRNA containing the TrkB 5' leader. Dissecting the 5' leader showed that the IRES is located in the exon common to all TrkB 5' leaders. Moreover, six regions ranging from 2 to 25 nt were identified that either promoted or inhibited IRES activity. Taken together, these results suggest that the 5' leader of the human TrkB mRNA contains multiple cis-elements that regulate internal initiation of translation and that this mechanism may contribute significantly to the translation of the TrkB mRNA in neuronal dendrites.

Present addresses: Angela Minic, Barbara Davis Center for Diabetes, 4200 E Ninth Ave Denver, CO 80262, USA

Elizabeth Amiott, Department of Biochemistry and Molecular Genetics, University of Colorado Health Sciences Center, Denver, CO 80262, USA


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