RNA-protein interactions within the 3 ' untranslated region of vimentin mRNA
| Autor: | Rebecca K. Shepherd, Zendra E. Zehner, W. Michael Holmes, Tzu Fun Fu, Mav Al-Ali, Joanna Gabryszuk |
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| Rok vydání: | 1997 |
| Předmět: |
Untranslated region
Molecular Sequence Data Sequence alignment RNA-binding protein Vimentin Biology Mice Species Specificity Cricetinae Consensus Sequence Genetics Intermediate Filament Protein Animals Humans RNA Messenger Messenger RNA Base Sequence Three prime untranslated region RNA RNA-Binding Proteins Molecular biology Cell Compartmentation Rats biology.protein Nucleic Acid Conformation Cattle Chickens Sequence Alignment HeLa Cells Research Article |
| Zdroj: | Nucleic acids research. 25(16) |
| ISSN: | 0305-1048 |
| Popis: | Several functions have been attributed to protein binding within the 3'untranslated region (3'UTR) of mRNA, including mRNA localization, stability, and translational repression. Vimentin is an intermediate filament protein whose 3'untranslated sequence is highly conserved between species. In order to identify sequences that might play a role in vimentin mRNA function, we synthesized32P-labeled RNA from different regions of vimentin's 3'UTR and assayed for protein binding with HeLa extracts using band shift assays. Sequences required for binding are contained within a region 61-114 nucleotides downstream of the stop codon, a region which is highly conserved from Xenopus to man. As judged by competition assays, binding is specific. Solution probing studies of 32P-labeled RNA with various nucleases and lead support a complex stem and loop structure for this region. Finally, UV cross-linking of the RNA-protein complex identifies an RNA binding protein of 46 kDa. Fractionation of a HeLa extract on a sizing column suggests that in addition to the 46 kDa protein, larger complexes containing additional protein(s) can be identified. Vimentin mRNA has been shown to be localized to the perinuclear region of the cytoplasm, possibly at sites of intermediate filament assembly. To date, all sequences required for localization of various mRNAs have been confined to the 3'UTR. Therefore, we hypothesize that this region and associated protein(s) might be important for vimentin mRNA function such as in localization. |
| Databáze: | OpenAIRE |
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