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M94A0081.TXT
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Document 0081
DOCN M94A0081
TI Differential DNA sequence specificity and regulation of HIV-1 enhancer
activity by cRel-RelA transcription factor.
DT 9412
AU Hansen SK; Guerrini L; Blasi F; Department of Genetics and Microbiol
Biology, University of; Milano, Italy.
SO J Biol Chem. 1994 Sep 2;269(35):22230-7. Unique Identifier : AIDSLINE
MED/94350977
AB 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.
DE Animal Base Sequence Binding Sites DNA, Viral/*METABOLISM *Enhancer
Elements (Genetics) Human HIV Long Terminal Repeat HIV-1/*GENETICS
Molecular Sequence Data NF-kappa B/*METABOLISM Polyomavirus
macacae/GENETICS Support, Non-U.S. Gov't Swine Trans-Activation
(Genetics) Transcription Factors/*METABOLISM Urokinase/GENETICS
JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).