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PMID: 1740106 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A novel complex between the p65 subunit of NF-kappa B and c-Rel binds to a DNA element involved in the phorbol ester induction of the human urokinase gene.

The EMBO journal ·Vol. 11 ·No. 1 ·1992-01-00 ·Pages 205-13

Hansen SK, Nerlov C, Zabel U, Verde P, Johnsen M, Baeuerle PA, Blasi F

Abstract

The NF-kappa B subunits, p50 and p65, have extensive sequence homology with the c-rel proto-oncogene and the Drosophila morphogen dorsal. It has recently been shown that in vitro translated c-Rel can bind to DNA and form a complex with p50. However, the conditions for DNA binding of c-Rel in vivo and its DNA sequence specificity have not been established. Here we report the identification a novel heterodimeric complex that binds to a kappa B-like, phorbol ester (TPA) responsive DNA sequence, 5'-GGGAAAGTAC-3', in the 5' flanking region of the human urokinase (uPA) gene. This sequence was shown to bind two protein complexes, LC and UC. LC was indistinguishable from NF-kappa B as it reacted with antibodies recognizing the p50 subunit of NF-kappa B, and was shown by UV crosslinking to contain the p50 and p65 subunits of NF-kappa B. UC, on the other hand, strongly reacted with anti-v-Rel, but not with the anti-p50 antibodies, and was shown by crosslinking to contain 75 kDa and 85 kDa protein-DNA adducts. The 75 kDa and the 85 kDa adducts could be immunoprecipitated only by anti-p65 and anti-c-Rel antibodies, respectively, showing that c-Rel formed a heterodimer with p65. Both protein complexes were present in inactive forms in HeLa cell cytosol, and their nuclear translocation was induced by TPA. DNA binding of UC and LC could, furthermore, be inhibited by I kappa B-alpha.(ABSTRACT TRUNCATED AT 250 WORDS)

MeSH Terms
Base Sequence Cell Line DNA-Binding Proteins/genetics Gene Expression Regulation/drug effects Humans Macromolecular Substances Molecular Sequence Data NF-kappa B/genetics Phorbol Esters/pharmacology Protein-Tyrosine Kinases/genetics Proto-Oncogene Mas Proto-Oncogene Proteins/genetics,immunology Proto-Oncogene Proteins c-rel Regulatory Sequences, Nucleic Acid/genetics Sequence Homology, Nucleic Acid Urokinase-Type Plasminogen Activator/genetics
Chemicals
DNA-Binding Proteins MAS1 protein, human Macromolecular Substances NF-kappa B Phorbol Esters Proto-Oncogene Mas Proto-Oncogene Proteins Proto-Oncogene Proteins c-rel Protein-Tyrosine Kinases Urokinase-Type Plasminogen Activator
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Hansen S K
University Institute of Microbiology, Copenhagen, Denmark.
Nerlov C
Zabel U
Verde P
Johnsen M
Baeuerle P A
Blasi F
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1992-01-00
Pages
205-13
Language
English
Region
England
NLM ID
8208664
PMCID
PMC556441
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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