Inhibition of RNA splicing at the Rous sarcoma virus src 3' splice site is mediated by an interaction between a negative cis element and a chicken embryo fibroblast nuclear factor

In permissive Rous sarcoma virus-infected chicken embryo fibroblasts (CEF), approximately equimolar amounts of env and src mRNAs are present. In nonpermissive mammalian cells, the src mRNA level is elevated and env mRNA level is reduced. A cis element in the region between the env gene and the src 3...

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Bibliographic Details
Published in:Journal of Virology 1995-08, Vol.69 (8), p.5068-5076
Main Authors: Amendt, B.A. (University of Iowa, Iowa City, IA.), Simpson, S.B, Stoltzfus, C.M
Format: Article
Language:English
Subjects:
ADN
AIDS/HIV
Animals
ARN MENSAJERO
ARN MESSAGER
Avian Sarcoma Viruses - genetics
Base Composition
Base Sequence
Chick Embryo
FIBROBLASTE
FIBROBLASTOS
GENE
GENES
Genes, env
Genes, src
HeLa Cells
Humans
Molecular Sequence Data
Nuclear Proteins - metabolism
Point Mutation
POLLITO
POUSSIN
PROTEINAS
PROTEINE
RNA Splicing
Rous sarcoma virus
SECUENCIA NUCLEICA
Sequence Deletion
SEQUENCE NUCLEIQUE
VIRUS DEL SARCOMA DE ROUS
VIRUS SARCOME DE ROUS
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Description
Summary:In permissive Rous sarcoma virus-infected chicken embryo fibroblasts (CEF), approximately equimolar amounts of env and src mRNAs are present. In nonpermissive mammalian cells, the src mRNA level is elevated and env mRNA level is reduced. A cis element in the region between the env gene and the src 3' splice site, which we have termed the suppressor of src splicing (SSS), acts specifically in CEF but not in human cells to reduce src mRNA levels. The splicing inhibition in CEF is not caused by a base-paired structure which is predicted to form between the SSS and the src 3' splice site. To further investigate the mechanism of the inhibition, we have used human HeLa cell nuclear extracts to compare in vitro the rates of splicing of RNA substrates containing the Rous sarcoma virus major 5' splice site and either the env or src 3' splice sites. We show that the src 3' splice site is used approximately fivefold more efficiently than the env 3' splice site. The efficiency of in vitro splicing at the src 3' splice site is specifically reduced by addition of CEF nuclear extract. The inhibition is dependent on the presence of the SSS element and can be abrogated by addition of competitor RNA. We propose that the SSS region represents a binding site for a negative-acting CEF splicing factor(s)
ISSN:0022-538X
1098-5514
DOI:10.1128/JVI.69.8.5068-5076.1995