| Mutational analysis of a viral RNA element that counteracts rapid RNA decay by interaction with the polyadenylate tail. | |
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MedLine Citation:
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PMID: 17563387 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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We previously demonstrated that the Kaposi's sarcoma-associated herpesvirus polyadenylated nuclear RNA contains a 79-nt cis-acting element, the ENE, which allows intronless polyadenylated transcripts to accumulate to high nuclear levels by protecting them from rapid degradation. We proposed a model based on the predicted structure of the ENE in which a U-rich internal loop hybridizes with the 3'-polyadenylate (polyA) tail to sequester it from exonucleolytic attack. We have tested this model by mutational analysis of the ENE. Point mutations in the predicted U-rich internal loop and in the flanking stems abolish the ENE's ability to (i) interact with the polyA tail, (ii) inhibit deadenylation in vitro, and (iii) stabilize transcripts in vivo. In all but one case, compensatory mutations in the flanking stems restore ENE activities, demonstrating the importance of these stems and uncovering a unique role for the loop-proximal G-C base pair in the lower stem. Increasing the U content of the U-rich internal loop surprisingly decreases stability in vivo but does not affect deadenylation in vitro, comparable to the effects of deleting certain "unstructured" regions of the ENE. Taken together, our data support the formation of the proposed ENE secondary structure in vivo and argue that the specific ENE structure inhibits rapid RNA decay in cis by engaging in a limited set of base-pairing interactions with the polyA tail. |
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Authors:
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Nicholas K Conrad; Mei-Di Shu; Katherine E Uyhazi; Joan A Steitz |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2007-06-11 |
Journal Detail:
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Title: Proceedings of the National Academy of Sciences of the United States of America Volume: 104 ISSN: 0027-8424 ISO Abbreviation: Proc. Natl. Acad. Sci. U.S.A. Publication Date: 2007 Jun |
Date Detail:
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Created Date: 2007-06-20 Completed Date: 2007-08-15 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 7505876 Medline TA: Proc Natl Acad Sci U S A Country: United States |
Other Details:
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Languages: eng Pagination: 10412-7 Citation Subset: IM |
Affiliation:
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Department of Molecular Biophysics and Biochemistry, Howard Hughes Medical Institute, Yale University, New Haven, CT 06536, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Base Pairing Base Sequence Hela Cells Humans Nucleic Acid Conformation Point Mutation Poly A / metabolism* Polyadenylation RNA Stability* RNA, Viral / genetics*, metabolism* Regulatory Sequences, Ribonucleic Acid / genetics* |
| Grant Support | |
ID/Acronym/Agency:
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5T32GM0705/GM/NIGMS NIH HHS; CA016038/CA/NCI NIH HHS; CA330006/CA/NCI NIH HHS; T32-CA09159-29/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/RNA, Viral; 0/Regulatory Sequences, Ribonucleic Acid; 24937-83-5/Poly A |
| Comments/Corrections | |
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