In eukaryotic cells, several mRNAs including those of c-myc and c-fos are localized to the perinuclear cytoplasm and associated with the cytoskeleton. The localization element of c-myc mRNA is present within its 3′UTR (3′-untranslated region) but the precise nature of this signal has remained unidentified. Chemical/enzymatic cleavage with RNases (ribonucleases) and lead have identified single-stranded and double-stranded regions in RNA transcripts of nucleotides 194–280 of the c-myc 3′UTR. Combined with computer predicted structure these results indicate that this region folds so that part of it forms a stem-loop structure. A mutation, that has been previously shown to prevent localization, leads to a different secondary RNA structure in this region as indicated by altered cleavage patterns. Competitive gel-retardation assays, using labelled transcripts corresponding to nucleotides 205–280 of c-myc 3′UTR, and fibroblast extracts revealed that the stem-loop region was sufficient for RNA–protein complex formation. In situ hybridization studies in cells transfected with reporter constructs, in which all or parts of the region corresponding to this stem-loop were linked to β-globin, indicated that this region was sufficient for localization and that deletion of the nucleotides corresponding to the proposed upper-stem or terminal loop prevented localization. Our hypothesis is that an AU-rich stem-loop structure within nt 222–267 in the c-myc 3′UTR forms the perinuclear localization signal. Bioinformatic analysis suggests that this signal shares features with 3′UTRs of other localized mRNAs and that these features may represent a conserved form of signal in mRNA localization mechanisms.
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December 2005
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Research Article|
December 06 2005
An AU-rich stem–loop structure is a critical feature of the perinuclear localization signal of c-myc mRNA
Hervé Chabanon;
Hervé Chabanon
1Institute for Cell and Molecular Biosciences, The Medical School, University of Newcastle-upon-Tyne NE2 4HH, U.K.
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Ian Mickleburgh;
Ian Mickleburgh
1Institute for Cell and Molecular Biosciences, The Medical School, University of Newcastle-upon-Tyne NE2 4HH, U.K.
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Brian Burtle;
Brian Burtle
1Institute for Cell and Molecular Biosciences, The Medical School, University of Newcastle-upon-Tyne NE2 4HH, U.K.
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Christopher Pedder;
Christopher Pedder
1Institute for Cell and Molecular Biosciences, The Medical School, University of Newcastle-upon-Tyne NE2 4HH, U.K.
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John Hesketh
John Hesketh
1
1Institute for Cell and Molecular Biosciences, The Medical School, University of Newcastle-upon-Tyne NE2 4HH, U.K.
1To whom correspondence should be addressed (email [email protected]).
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Publisher: Portland Press Ltd
Received:
May 17 2005
Revision Received:
July 21 2005
Accepted:
July 26 2005
Accepted Manuscript online:
July 26 2005
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2005
Biochem J (2005) 392 (3): 475–483.
Article history
Received:
May 17 2005
Revision Received:
July 21 2005
Accepted:
July 26 2005
Accepted Manuscript online:
July 26 2005
Citation
Hervé Chabanon, Ian Mickleburgh, Brian Burtle, Christopher Pedder, John Hesketh; An AU-rich stem–loop structure is a critical feature of the perinuclear localization signal of c-myc mRNA. Biochem J 15 December 2005; 392 (3): 475–483. doi: https://doi.org/10.1042/BJ20050812
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