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Identification of cis-acting sequences responsible for phorbol ester induction of human serum amyloid A gene expression via a nuclear factor kB-like transcription factor

Journal Article · · Mol. Cell. Biol.; (United States)
DOI:https://doi.org/10.1128/MCB.9.5.1908· OSTI ID:5823305
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The authors have analyzed the 5'-flanking region of one of the genes coding for the human acute-phase protein, serum amyloid A (SAA). They found that SAA mRNA could be increased fivefold in transfected cells by treatment with phorbol 12-myristate 13-acetate (PMA). To analyze this observation further, they placed a 265-base-pair 5' SAA fragment upstream of the reporter chloramphenicol acetyltransferase (CAT) gene and transfected this construct into HeLa cells. PMA treatment of these transient transfectants resulted in increased CAT expression. Nuclear proteins from PMA-treated HeLa cells bound to this DNA fragment, and methylation interference analysis showed that the binding was specific to the sequence GGGACTTTCC (between -82 and -91), a sequence previously described by others as the binding site for the nuclear factor NF/kappa/B. In a cotransfection competition experiment, they could abolish PMA-induced CAT activity by using cloned human immunodeficiency virus long-terminal-repeat DNA containing the NF/kappa/B-binding sequence. The same long-terminal-repeat DNA containing mutant NF/kappa/B-binding sequences did not affect CAT expression, which suggested that binding by an NF/kappa/B-like factor is required for increased SAA transcription.
Research Organization:
MRC Clinical Research Centre, Harrow, Middlesex HA1 3UJ (GB); AFRC Physiology and Genetics Research Station, Roslin, Midlothian EH25 9PS (GB)
OSTI ID:
5823305
Journal Information:
Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 9:5; ISSN MCEBD
Country of Publication:
United States
Language:
English

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Related Subjects

550200* -- Biochemistry
550400 -- Genetics
59 BASIC BIOLOGICAL SCIENCES
AIDS VIRUS
AMINO ACID SEQUENCE
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
BIOSYNTHESIS
CARCINOGENS
CHEMICAL REACTIONS
CHLORAMPHENICOL
CLONING
DNA HYBRIDIZATION
DNA-CLONING
DRUGS
ENZYME INDUCTION
ENZYMES
ESTERS
GENE REGULATION
GENE REPRESSORS
GENES
GENETIC ENGINEERING
HELA CELLS
HYBRIDIZATION
MESSENGER-RNA
METHYLATION
MICROORGANISMS
MOLECULAR BIOLOGY
MOLECULAR STRUCTURE
MUTANTS
NUCLEIC ACIDS
NUCLEOPROTEINS
ORGANIC COMPOUNDS
PARASITES
PHENOTYPE
PHORBOL ESTERS
PROTEINS
RNA
RNA PROCESSING
SYNTHESIS
TRANSCRIPTION FACTORS
TRANSFERASES
VIRUSES