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Nucleic Acids Research Advance Access published online on September 14, 2009
Nucleic Acids Research, doi:10.1093/nar/gkp713
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© The Author(s) 2009. Published by Oxford University Press.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

RNA

Translation of the FMR1 mRNA is not influenced by AGG interruptions

Anna L. Ludwig1, Christopher Raske1, Flora Tassone1,2, Dolores Garcia-Arocena1, John W. Hershey1 and Paul J. Hagerman1,*

1Department of Biochemistry and Molecular Medicine, School of Medicine, University of California Davis, 4303 Tupper Hall, Davis, CA 95616 and 2MIND Institute, University of California Davis, Health System, Sacramento, CA, USA

*To whom correspondence should be addressed. Tel: +1 530 754 7266; Fax: +1 530 754 7269; Email: pjhagerman{at}ucdavis.edu

Received July 15, 2009. Revised August 12, 2009. Accepted August 13, 2009.

The fragile X mental retardation 1 (FMR1) gene contains a CGG-repeat element within its 5' untranslated region (5'UTR) which, for alleles with more than ~40 repeats, increasingly affects both transcription (up-regulation) and translation (inhibition) of the repeat-containing RNA with increasing CGG-repeat length. Translational inhibition is thought to be due to impaired ribosomal scanning through the CGG-repeat region, which is postulated to form highly stable secondary/tertiary structure. One striking difference between alleles in the premutation range (55–200 CGG repeats) and those in the normal range (<~40 repeats) is the reduced number/absence of ‘expansion stabilizing’ AGG interruptions in the larger alleles. Such interruptions, which generally occur every 9–11 repeats in normal alleles, are thought to disrupt the extended CGG-repeat hairpin structure, thus facilitating translational initiation. To test this hypothesis, we have measured the translational efficiency of CGG-repeat mRNAs with 0–2 AGG interruptions, both in vitro (rabbit reticulocyte lysates) and in cell culture (HEK-293 cells). We demonstrate that the AGG interruptions have no detectable influence on translational efficiency in either a cell-free system or cell culture, indicating that any AGG-repeat-induced alterations in secondary/tertiary structure, if present, do not involve the rate-limiting step(s) in translational initiation.


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