Document Detail


Differential DNA sequence specificity and regulation of HIV-1 enhancer activity by cRel-RelA transcription factor.
MedLine Citation:
PMID:  8071349     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The cRel-RelA and NF-kappa B (p50-RelA) transcription factors bind to a kappa B-like sequence termed Rel-related proteins binding element localized in the regulatory region of the human urokinase plasminogen activator (uPA) gene. This sequence is highly conserved in murine and porcine uPA genes where it retained the ability to associate with cRel-RelA. On the other hand, NF-kappa B binding was obtained with the human and porcine elements only. Methylation interference analysis showed that NF-kappa B and cRel-RelA had identical interference patterns. Mutational analysis showed that DNA binding was highly sensitive to mutations within the decameric Rel-related proteins binding element core site. However, alterations of nucleotides flanking the decameric IgK-kappa B motif, which preferentially associated with NF-kappa B, resulted in high affinity cRel-RelA binding both in vitro and in vivo. These data demonstrate that NF-kappa B and cRel-RelA have overlapping but distinct DNA sequence specificities. Bandshift analysis with HeLa and Jurkat cell extracts or with in vitro translated proteins revealed that the SV40-, HIV-1-, and interleukin-2 receptor alpha subunit kappa B elements efficiently associated with cRel-RelA, suggesting that this heterodimer may be involved in the regulation of several genes. Cotransfection studies of HIV-1 long terminal repeat-chloramphenicol acetyltransferase reporter DNA with RelA, cRel, and p50 expression vectors were performed in COS7 and U293 cells to analyze the ability of cRel-RelA to regulate HIV-1 enhancer activity. In vivo formation of the cRel-RelA complex resulted in specific stimulation of the viral enhancer at a level comparable with that obtained with NF-kappa B. These data suggest that activation of cellular cRel-RelA may play a critical role in the regulation of HIV-1 enhancer activity.
Authors:
S K Hansen; L Guerrini; F Blasi
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  269     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1994 Sep 
Date Detail:
Created Date:  1994-09-29     Completed Date:  1994-09-29     Revised Date:  2012-02-27    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  22230-7     Citation Subset:  IM; X    
Affiliation:
Department of Genetics and Microbiol Biology, University of Milano, Italy.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
Binding Sites
DNA, Viral / metabolism*
Enhancer Elements, Genetic*
HIV Long Terminal Repeat
HIV-1 / genetics*
Humans
Molecular Sequence Data
NF-kappa B / metabolism*
Simian virus 40 / genetics
Swine
Transcription Factor RelA
Transcription Factors / metabolism*
Transcriptional Activation
Urokinase-Type Plasminogen Activator / genetics
Grant Support
ID/Acronym/Agency:
168//Telethon
Chemical
Reg. No./Substance:
0/DNA, Viral; 0/NF-kappa B; 0/Transcription Factor RelA; 0/Transcription Factors; EC 3.4.21.73/Urokinase-Type Plasminogen Activator

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