An RNA element at the 5'-end of the poliovirus genome functions as a general promoter for RNA synthesis.

RNA structures present throughout RNA virus genomes serve as scaffolds to organize multiple factors involved in the initiation of RNA synthesis. Several of these RNA elements play multiple roles in the RNA replication pathway. An RNA structure formed around the 5'- end of the poliovirus genomic...

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Main Authors: Dorothee A Vogt, Raul Andino
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2010-06-01
Series:PLoS Pathogens
Online Access:https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1000936&type=printable
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author Dorothee A Vogt
Raul Andino
author_facet Dorothee A Vogt
Raul Andino
author_sort Dorothee A Vogt
collection DOAJ
description RNA structures present throughout RNA virus genomes serve as scaffolds to organize multiple factors involved in the initiation of RNA synthesis. Several of these RNA elements play multiple roles in the RNA replication pathway. An RNA structure formed around the 5'- end of the poliovirus genomic RNA has been implicated in the initiation of both negative- and positive-strand RNA synthesis. Dissecting the roles of these multifunctional elements is usually hindered by the interdependent nature of the viral replication processes and often pleiotropic effects of mutations. Here, we describe a novel approach to examine RNA elements with multiple roles. Our approach relies on the duplication of the RNA structure so that one copy is dedicated to the initiation of negative-strand RNA synthesis, while the other mediates positive-strand synthesis. This allows us to study the function of the element in promoting positive-strand RNA synthesis, independently of its function in negative-strand initiation. Using this approach, we demonstrate that the entire 5'-end RNA structure that forms on the positive-strand is required for initiation of new positive-strand RNAs. Also required to initiate positive-strand RNA synthesis are the binding sites for the viral polymerase precursor, 3CD, and the host factor, PCBP. Furthermore, we identify specific nucleotide sequences within "stem a" that are essential for the initiation of positive-strand RNA synthesis. These findings provide direct evidence for a trans-initiation model, in which binding of proteins to internal sequences of a pre-existing positive-strand RNA affects the synthesis of subsequent copies of that RNA, most likely by organizing replication factors around the initiation site.
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spelling doaj-art-8e2a42efdbdf4d179d02862a9006b4322025-08-20T03:07:20ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742010-06-0166e100093610.1371/journal.ppat.1000936An RNA element at the 5'-end of the poliovirus genome functions as a general promoter for RNA synthesis.Dorothee A VogtRaul AndinoRNA structures present throughout RNA virus genomes serve as scaffolds to organize multiple factors involved in the initiation of RNA synthesis. Several of these RNA elements play multiple roles in the RNA replication pathway. An RNA structure formed around the 5'- end of the poliovirus genomic RNA has been implicated in the initiation of both negative- and positive-strand RNA synthesis. Dissecting the roles of these multifunctional elements is usually hindered by the interdependent nature of the viral replication processes and often pleiotropic effects of mutations. Here, we describe a novel approach to examine RNA elements with multiple roles. Our approach relies on the duplication of the RNA structure so that one copy is dedicated to the initiation of negative-strand RNA synthesis, while the other mediates positive-strand synthesis. This allows us to study the function of the element in promoting positive-strand RNA synthesis, independently of its function in negative-strand initiation. Using this approach, we demonstrate that the entire 5'-end RNA structure that forms on the positive-strand is required for initiation of new positive-strand RNAs. Also required to initiate positive-strand RNA synthesis are the binding sites for the viral polymerase precursor, 3CD, and the host factor, PCBP. Furthermore, we identify specific nucleotide sequences within "stem a" that are essential for the initiation of positive-strand RNA synthesis. These findings provide direct evidence for a trans-initiation model, in which binding of proteins to internal sequences of a pre-existing positive-strand RNA affects the synthesis of subsequent copies of that RNA, most likely by organizing replication factors around the initiation site.https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1000936&type=printable
spellingShingle Dorothee A Vogt
Raul Andino
An RNA element at the 5'-end of the poliovirus genome functions as a general promoter for RNA synthesis.
PLoS Pathogens
title An RNA element at the 5'-end of the poliovirus genome functions as a general promoter for RNA synthesis.
title_full An RNA element at the 5'-end of the poliovirus genome functions as a general promoter for RNA synthesis.
title_fullStr An RNA element at the 5'-end of the poliovirus genome functions as a general promoter for RNA synthesis.
title_full_unstemmed An RNA element at the 5'-end of the poliovirus genome functions as a general promoter for RNA synthesis.
title_short An RNA element at the 5'-end of the poliovirus genome functions as a general promoter for RNA synthesis.
title_sort rna element at the 5 end of the poliovirus genome functions as a general promoter for rna synthesis
url https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1000936&type=printable
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