Liver enriched transcription factor
First Claim
1. A competitive assay for binding activity to recombinant HNF-4 receptors, comprising:
- (a) reacting a suspected ligand with a recombinant HNF-4 receptor in the presence of a known quantity of a labeled competing ligand; and
(b) comparing the amount of bound label in the presence of the suspected ligand to that observed in its absence, wherein the HNF-4 receptor is a chimeric receptor comprising the ligand binding domain of a HNF-4 receptor and a DNA binding domain of another receptor which has a known ligand.
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Abstract
HNF-4 (hepatocyte nuclear factor 4) is a protein enriched in liver extracts that binds to sites required for the transcription of the transthyretin (TTR) and apolipoprotein CIII (apoCIII) genes (Costa et al., 1989; Costa et al., 1990; Leff et al., 1989). We have purified HNF-4 protein (54 kD) and isolated a cDNA clone encoding the protein. HNF-4 is a member of the steroid hormone receptor superfamily with an unusual amino acid in the conserved “knuckle” of the first zinc finger (DGCKG). This and the fact that HNF-4 does not bind significantly to estrogen, thyroid hormone or glucocorticoid response elements indicate that HNF-4 may represent a new subfamily. HNF-4 binds to its recognition site as a dimer and activates transcription in a sequence-specific fashion in nonhepatic (HeLa) cells. HNF-4 mRNA is present in kidney and intestine as well as liver but is absent in other tissues. DNA binding data suggest that HNF-4 could be identical to liver factor A1 (LF-A1), a factor previously shown to regulate the transcription of the α-1 antitrypsin, apolipoprotein A1 and pyruvate kinase genes.
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Citations
5 Claims
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1. A competitive assay for binding activity to recombinant HNF-4 receptors, comprising:
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(a) reacting a suspected ligand with a recombinant HNF-4 receptor in the presence of a known quantity of a labeled competing ligand; and
(b) comparing the amount of bound label in the presence of the suspected ligand to that observed in its absence, wherein the HNF-4 receptor is a chimeric receptor comprising the ligand binding domain of a HNF-4 receptor and a DNA binding domain of another receptor which has a known ligand. - View Dependent Claims (2, 3, 4, 5)
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Specification