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M9480857.TXT
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1994-09-05
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Document 0857
DOCN M9480857
TI The upregulation of the promoter gene encoding the p105/p50 subunit of
NF-kB by HIV is dependent upon one specific kB motif.
DT 9410
AU Ten R; MacMorran W; Rolli V; Israel A; Paya C; Division of Experimental
Pathology, Mayo Clinic, Rochester, MN; 55905.
SO Abstr Gen Meet Am Soc Microbiol. 1994;94:483 (abstract no. T-5). Unique
Identifier : AIDSLINE ASM94/94313088
AB Our previous cloning and sequencing of the promoter of the gene encoding
the p50 subunit of NF-kB indicated that it is upregulated by NF-kB.
Furthermore, we have shown that in persistently HIV-infected monocytic
cells, which contain increased NF-kB activity, the transcriptional
regulation of this promoter is increased. We have further characterized
the different regulatory sequences of the p105/p50 promoter that could
potentially participate in its autoregulation. One AP-1, four NF-kB like
and one HIP-1 motifs are present in this promoter. Gel shift analysis
using nuclear extracts from monocytic cells unstimulated or treated with
LPS or HIV-infected demonstrated specific binding proteins to the AP-1
site, to the three of the four kB sites, but not to the HIP-1 motif. One
of the three kB sites (kB4) bound NF-kB complexes with higher affinity
than the rest. Deletion and mutation constructs of the p105 promoter
cloned upstream a luciferase reporter gene followed by transfection into
monocytic cells was performed. The AP-1, kB1, kB2, kB3, and HIP-1
regions were found to be dispensable for the basal and LPS-inducible
transcriptional activity of the promoter in uninfected and HIV-infected
monocytes. Only the kB4 site was identified as essential in the
regulation of the p105 promoter. These results suggest that the p105/p50
promoter is solely regulated by NF-kB complexes binding with high
affinity to only one of the kB motifs in monocytic cells.
DE Binding Sites *Gene Expression Regulation, Viral Human HIV/*GENETICS
Lipopolysaccharides/PHARMACOLOGY Luciferase/BIOSYNTHESIS
Monocytes/DRUG EFFECTS/*METABOLISM/MICROBIOLOGY NF-kappa
B/BIOSYNTHESIS/*GENETICS/METABOLISM *Promoter Regions (Genetics)
Recombinant Proteins/BIOSYNTHESIS Transcription, Genetic Transfection
MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).