Document Detail


Binding sites for Rev and ASF/SF2 map to a 55-nucleotide purine-rich exonic element in equine infectious anemia virus RNA.
MedLine Citation:
PMID:  11278454     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The equine infectious anemia virus (EIAV) Rev protein (ERev) negatively regulates its own synthesis by inducing alternative splicing of its mRNA. This bicistronic mRNA contains four exons; exons 1 and 2 encode Tat, and exons 3 and 4 encode Rev. When Rev is expressed, exon 3 is skipped to produce an mRNA that contains only exons 1, 2, and 4. The interaction of ERev with its cis-acting RNA response element, the RRE, is also essential for nuclear export of intron-containing viral mRNAs that encode structural and enzymatic gene products. The primary ERev binding site and the manner in which ERev interacts with RNA or cellular proteins to exert its regulatory function have not been defined. We have performed in vitro RNA binding experiments to show that recombinant ERev binds to a 55-nucleotide, purine-rich tract proximal to the 5' splice site of exon 3. Because of its proximity to the 5' splice site and since it contains elements related to consensus exonic splicing enhancer sequences, we asked whether cellular proteins recognize the EIAV RRE. The cellular protein, ASF/SF2, a member of the serine- and arginine-rich family of splicing factors (SR proteins) bound to repeated sequences within the 55-nucleotide RRE region. Electrophoretic mobility shift and UV cross-linking experiments indicated that ERev and SR proteins bind simultaneously to the RRE. Furthermore, in vitro protein-protein interaction studies revealed an association between ERev and SR proteins. These data suggest that EIAV Rev-induced exon skipping observed in vivo may be initiated by simultaneous binding of Rev and SR proteins to the RRE that alter the subsequent assembly or catalytic activity of the spliceosomal complex.
Authors:
Chung H; D Derse
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Publication Detail:
Type:  Journal Article     Date:  2001-03-16
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  276     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2001 Jun 
Date Detail:
Created Date:  2001-05-30     Completed Date:  2001-07-26     Revised Date:  2011-09-27    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  18960-7     Citation Subset:  IM    
Affiliation:
Basic Research Laboratory, NCI-Frederick, National Institutes of Health, Frederick, Maryland 21702.
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MeSH Terms
Descriptor/Qualifier:
Alternative Splicing
Base Sequence
Binding Sites
Catalysis
Cell Nucleus / metabolism
Dimerization
Dose-Response Relationship, Drug
Exons
Gene Products, rev / chemistry,  metabolism*
Hela Cells
Humans
Infectious Anemia Virus, Equine / genetics*,  metabolism*
Introns
Molecular Sequence Data
Nuclear Proteins / chemistry,  metabolism*
Plasmids / metabolism
Precipitin Tests
Protein Binding
RNA / metabolism
RNA Processing, Post-Transcriptional
RNA, Messenger / metabolism
Sequence Homology, Nucleic Acid
Spliceosomes / metabolism
Ultraviolet Rays
Chemical
Reg. No./Substance:
0/Gene Products, rev; 0/Nuclear Proteins; 0/RNA, Messenger; 170974-22-8/serine-arginine-rich splicing proteins; 63231-63-0/RNA

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