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 |
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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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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.</description><subject>Base Sequence</subject><subject>Cell Line</subject><subject>Ebolavirus - genetics</subject><subject>Ebolavirus - physiology</subject><subject>Genetic Engineering</subject><subject>Genome, Viral</subject><subject>Humans</subject><subject>Models, Molecular</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis, Site-Directed</subject><subject>Nucleic Acid Conformation</subject><subject>RNA, Viral - chemistry</subject><subject>RNA, Viral - genetics</subject><subject>Thermodynamics</subject><subject>Virus Replication</subject><issn>0042-6822</issn><issn>1096-0341</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1Lw0AQhhdRbK3-BGVPoofofky26UlKqR9QFNSel2R3gitptu4mhf57E1v0KAzMHJ6Zl3kIOefshjOubt8YA5GoTIgrJq4ZYxwSOCBDziYqYRL4IRn-IgNyEuNnB8F4zI7JgAsFgk1gSJbTOq-20UXqS9p8IA2-wn5eB7TONGjp6_OURjS-tnnY0tiE1jRtwEhdTeeFr3K6caGNNOC6ciZvnK9PyVGZVxHP9n1Elvfz99ljsnh5eJpNF4kBgCYxivNcpWCzEpRiBZ_wLDU2BVSWp1LYtOACwRhppSwsjOWkkCAVlJlEZkCOyOXu7jr4rxZjo1cuGqyqvEbfRs2zcbckeAemO9AEH2PAUq-DW3X_aM5071P_-NS9LM266n3qPuBiH9AWK7R_W3uBHXC3A7B7c-Mw6Ggc1qaTF9A02nr3T8Q3ejaEHg</recordid><startdate>20030215</startdate><enddate>20030215</enddate><creator>Crary, Sharon M</creator><creator>Towner, Jonathan S</creator><creator>Honig, Jessica E</creator><creator>Shoemaker, Trevor R</creator><creator>Nichol, Stuart T</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>7U9</scope><scope>H94</scope></search><sort><creationdate>20030215</creationdate><title>Analysis of the role of predicted RNA secondary structures in Ebola virus replication</title><author>Crary, Sharon M ; Towner, Jonathan S ; Honig, Jessica E ; Shoemaker, Trevor R ; Nichol, Stuart T</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c444t-c611a654d8f4660b19185cd54e6d1532d5b12e4cc3d33bd4739b34364f83e0c43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Base Sequence</topic><topic>Cell Line</topic><topic>Ebolavirus - genetics</topic><topic>Ebolavirus - physiology</topic><topic>Genetic Engineering</topic><topic>Genome, Viral</topic><topic>Humans</topic><topic>Models, Molecular</topic><topic>Molecular Sequence Data</topic><topic>Mutagenesis, Site-Directed</topic><topic>Nucleic Acid Conformation</topic><topic>RNA, Viral - chemistry</topic><topic>RNA, Viral - genetics</topic><topic>Thermodynamics</topic><topic>Virus Replication</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Crary, Sharon M</creatorcontrib><creatorcontrib>Towner, Jonathan S</creatorcontrib><creatorcontrib>Honig, Jessica E</creatorcontrib><creatorcontrib>Shoemaker, Trevor R</creatorcontrib><creatorcontrib>Nichol, Stuart T</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><jtitle>Virology (New York, N.Y.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Crary, Sharon M</au><au>Towner, Jonathan S</au><au>Honig, Jessica E</au><au>Shoemaker, Trevor R</au><au>Nichol, Stuart T</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Analysis of the role of predicted RNA secondary structures in Ebola virus replication</atitle><jtitle>Virology (New York, N.Y.)</jtitle><addtitle>Virology</addtitle><date>2003-02-15</date><risdate>2003</risdate><volume>306</volume><issue>2</issue><spage>210</spage><epage>218</epage><pages>210-218</pages><issn>0042-6822</issn><eissn>1096-0341</eissn><abstract>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. 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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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