Analysis of the role of predicted RNA secondary structures in Ebola virus replication

Thermodynamic modeling of Ebola viral RNA predicts the formation of RNA stem-loop structures at the 3′ and 5′ termini and panhandle structures between the termini of the genomic (or antigenomic) RNAs. Sequence analysis showed a high degree of identity among Ebola Zaire, Sudan, Reston, and Cote d’Ivo...

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Veröffentlicht in:Virology (New York, N.Y.) N.Y.), 2003-02, Vol.306 (2), p.210-218
Hauptverfasser: Crary, Sharon M, Towner, Jonathan S, Honig, Jessica E, Shoemaker, Trevor R, Nichol, Stuart T
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container_start_page 210
container_title Virology (New York, N.Y.)
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creator Crary, Sharon M
Towner, Jonathan S
Honig, Jessica E
Shoemaker, Trevor R
Nichol, Stuart T
description Thermodynamic modeling of Ebola viral RNA predicts the formation of RNA stem-loop structures at the 3′ and 5′ termini and panhandle structures between the termini of the genomic (or antigenomic) RNAs. Sequence analysis showed a high degree of identity among Ebola Zaire, Sudan, Reston, and Cote d’Ivoire subtype viruses in their 3′ and 5′ termini (18 nucleotides in length) and within a second region (internal by approximately 20 nucleotides). While base pairing of the two conserved regions could lead to the formation of the base of the putative stem-loop or panhandle structures, the intervening sequence variation altered the predictions for the rest of the structures. Using an in vivo minigenome replication system, we engineered mutations designed to disrupt potential base pairing in the viral RNA termini. Analysis of these variants by screening for enhanced green fluorescent protein reporter expression and by quantitation of minigenomic RNA levels demonstrated that the upper portions of the putative panhandle and 3′ genomic structures can be destabilized without affecting virus replication.
doi_str_mv 10.1016/S0042-6822(02)00014-4
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subjects Base Sequence
Cell Line
Ebolavirus - genetics
Ebolavirus - physiology
Genetic Engineering
Genome, Viral
Humans
Models, Molecular
Molecular Sequence Data
Mutagenesis, Site-Directed
Nucleic Acid Conformation
RNA, Viral - chemistry
RNA, Viral - genetics
Thermodynamics
Virus Replication
title Analysis of the role of predicted RNA secondary structures in Ebola virus replication
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