| Endonuclease substrate selectivity characterized with full-length PA of influenza A virus polymerase. | |
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MedLine Citation:
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PMID: 22841552 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The influenza A polymerase is a heterotrimer which transcribes viral mRNAs and replicates the viral genome. To initiate synthesis of mRNA, the polymerase binds a host pre-mRNA and cleaves a short primer downstream of the 5' end cap structure. The N-terminal domain of PA has been demonstrated to have endonuclease activity in vitro. Here we sought to better understand the biochemical nature of the PA endonuclease by developing an improved assay using full-length PA protein. This full-length protein is active against both RNA and DNA in a cap-independent manner and can use several different divalent cations as cofactors, which affects the secondary structure of the full-length PA. Our in vitro assay was also able to demonstrate the minimal substrate size and sequence selectivity of the PA protein, which is crucial information for inhibitor design. Finally, we confirmed the observed endonuclease activity of the full-length PA with a FRET-based assay. |
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Authors:
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Erin Noble; Andrew Cox; Jerome Deval; Baek Kim |
Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2012-07-28 |
Journal Detail:
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Title: Virology Volume: 433 ISSN: 1096-0341 ISO Abbreviation: Virology Publication Date: 2012 Nov |
Date Detail:
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Created Date: 2012-09-17 Completed Date: 2013-01-04 Revised Date: 2013-05-09 |
Medline Journal Info:
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Nlm Unique ID: 0110674 Medline TA: Virology Country: United States |
Other Details:
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Languages: eng Pagination: 27-34 Citation Subset: IM |
Copyright Information:
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Copyright © 2012 Elsevier Inc. All rights reserved. |
Affiliation:
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Department of Microbiology and Immunology, University of Rochester, Rochester, NY 14642, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Biological Assay Cations, Divalent Coenzymes / metabolism Endonucleases / chemistry, genetics, metabolism* Fluorescence Resonance Energy Transfer Humans Influenza A virus / enzymology*, genetics Magnesium / metabolism Manganese / metabolism Protein Multimerization Protein Structure, Secondary Protein Structure, Tertiary RNA Caps / chemistry*, genetics RNA Replicase / chemistry, genetics, metabolism* RNA, Viral / chemistry*, genetics Recombinant Proteins / chemistry, genetics, metabolism Substrate Specificity Viral Proteins / chemistry, genetics, metabolism* |
| Grant Support | |
ID/Acronym/Agency:
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266200700008C//PHS HHS; T32 DE007202/DE/NIDCR NIH HHS; T32 DE007202/DE/NIDCR NIH HHS; T32GM07356/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Cations, Divalent; 0/Coenzymes; 0/PA protein, influenza viruses; 0/RNA Caps; 0/RNA, Viral; 0/Recombinant Proteins; 0/Viral Proteins; 7439-95-4/Magnesium; 7439-96-5/Manganese; EC 2.7.7.48/RNA Replicase; EC 3.1.-/Endonucleases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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