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Identification and characterization of extensive intra-molecular associations between 3′-UTRs and their ORFs
During eukaryotic translation, mRNAs may form intra-molecular interactions between distant domains. The 5′-cap and the polyA tail were shown to interact through their associated proteins, and this can induce physical compaction of the mRNA in vitro. However, the stability of this intra-molecular ass...
Autores principales: | , , |
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Formato: | Texto |
Lenguaje: | English |
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Oxford University Press
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2588509/ https://www.ncbi.nlm.nih.gov/pubmed/18948291 http://dx.doi.org/10.1093/nar/gkn754 |
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author | Eldad, Naama Yosefzon, Yahav Arava, Yoav |
author_facet | Eldad, Naama Yosefzon, Yahav Arava, Yoav |
author_sort | Eldad, Naama |
collection | PubMed |
description | During eukaryotic translation, mRNAs may form intra-molecular interactions between distant domains. The 5′-cap and the polyA tail were shown to interact through their associated proteins, and this can induce physical compaction of the mRNA in vitro. However, the stability of this intra-molecular association in translating mRNAs and whether additional contacts exist in vivo are largely unknown. To explore this, we applied a novel approach in which several endogenous polysomal mRNAs from Saccharomyces cerevisiae were cleaved near their stop codon and the resulting 3′-UTR fragments were tested either for co-sedimentation or co-immunoprecipitation (co-IP) with their ORFs. In all cases a significant fraction of the 3′-UTR fragments sedimented similarly to their ORF-containing fragments, yet the extent of co-sedimentation differed between mRNAs. Similar observations were obtained by a co-IP assay. Interestingly, various treatments that are expected to interfere with the cap to polyA interactions had no effect on the co-sedimentation pattern. Moreover, the 3′-UTR appeared to co-sediment with different regions from within the ORF. Taken together, these results indicate extensive physical associations between 3′-UTRs and their ORFs that vary between genes. This implies that polyribosomal mRNAs are in a compact configuration in vivo. |
format | Text |
id | pubmed-2588509 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-25885092009-03-04 Identification and characterization of extensive intra-molecular associations between 3′-UTRs and their ORFs Eldad, Naama Yosefzon, Yahav Arava, Yoav Nucleic Acids Res RNA During eukaryotic translation, mRNAs may form intra-molecular interactions between distant domains. The 5′-cap and the polyA tail were shown to interact through their associated proteins, and this can induce physical compaction of the mRNA in vitro. However, the stability of this intra-molecular association in translating mRNAs and whether additional contacts exist in vivo are largely unknown. To explore this, we applied a novel approach in which several endogenous polysomal mRNAs from Saccharomyces cerevisiae were cleaved near their stop codon and the resulting 3′-UTR fragments were tested either for co-sedimentation or co-immunoprecipitation (co-IP) with their ORFs. In all cases a significant fraction of the 3′-UTR fragments sedimented similarly to their ORF-containing fragments, yet the extent of co-sedimentation differed between mRNAs. Similar observations were obtained by a co-IP assay. Interestingly, various treatments that are expected to interfere with the cap to polyA interactions had no effect on the co-sedimentation pattern. Moreover, the 3′-UTR appeared to co-sediment with different regions from within the ORF. Taken together, these results indicate extensive physical associations between 3′-UTRs and their ORFs that vary between genes. This implies that polyribosomal mRNAs are in a compact configuration in vivo. Oxford University Press 2008-12 2008-10-23 /pmc/articles/PMC2588509/ /pubmed/18948291 http://dx.doi.org/10.1093/nar/gkn754 Text en © 2008 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | RNA Eldad, Naama Yosefzon, Yahav Arava, Yoav Identification and characterization of extensive intra-molecular associations between 3′-UTRs and their ORFs |
title | Identification and characterization of extensive intra-molecular associations between 3′-UTRs and their ORFs |
title_full | Identification and characterization of extensive intra-molecular associations between 3′-UTRs and their ORFs |
title_fullStr | Identification and characterization of extensive intra-molecular associations between 3′-UTRs and their ORFs |
title_full_unstemmed | Identification and characterization of extensive intra-molecular associations between 3′-UTRs and their ORFs |
title_short | Identification and characterization of extensive intra-molecular associations between 3′-UTRs and their ORFs |
title_sort | identification and characterization of extensive intra-molecular associations between 3′-utrs and their orfs |
topic | RNA |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2588509/ https://www.ncbi.nlm.nih.gov/pubmed/18948291 http://dx.doi.org/10.1093/nar/gkn754 |
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