NF-κB activating gene
Provided are proteins having NF-κB activity, which are used for diagnosing, treating or preventing diseases associated with the excessive activation or inhibition of NF-κB. Using plasmid pNFκB-Luc, cDNA encoding a protein capable of activating NF-κB has been cloned from a cDNA library constructed from human lung fibroblasts, and the DNA sequence and the deduced amino acid sequence determined. The protein, the DNA encoding the protein, a recombinant vector containing the DNA, and a transformant containing the recombinant vector are useful for screening a substance inhibiting or promoting NF-κB activation.
1. An isolated polynucleotide which comprises a nucleotide sequence encoding a protein that activates NF-κB and selected from the group consisting of:
(a) a protein which comprises an amino acid sequence represented by SEQ ID NO: 87.
2. An isolated polynucleotide comprising a polynucleotide sequence selected from the group consisting of:
(a) a polynucleotide sequence represented by SEQ ID NO: 88;
(b) a polynucleotide sequence encoding a protein that activates NF-κB and hybridizing under stringent conditions with a polynucleotide having a polynucleotide sequence complementary to the polynucleotide sequence of (a).
3. An isolated polynucleotide comprising a polynucleotide sequence selected from the group consisting of:
(a) a nucleotide sequence represented by a coding region in SEQ ID NO: 88;
(b) a nucleotide sequence encoding a protein that activates NF-κB and hybridizing under stringent conditions with a polynucleotide having a polynucleotide sequence complementary to the polynucleotide sequence of (a).
4. An isolated polynucleotide comprising a nucleotide sequence which encodes a protein that activates NF-κB and has at least 95% identity to the polynucleotide sequence according to claim 1 over the entire length thereof.
5. An isolated polynucleotide comprising a nucleotide sequence which encodes a protein that activates NF-κB and has at least 95% identity to the polynucleotide sequence according to claim 2 or 3 over the entire length thereof.
6. A recombinant vector which comprises a polynucleotide according to any one of claims 1 to 4 .
7. A transformed cell which comprises the recombinant vector according to claim 6 .
8. A process for producing a protein comprising,
(a) culturing a transformed cell comprising the isolated polynucleotide according to any one of claims 1 to 4 under conditions providing expression of the encoded protein; and
(b) recovering the protein from the culture product.
9. A method for screening compounds in respect of activity to inhibit or promote NF-κB activation, which comprises the steps of:
(a) providing a cell with a gene comprising a polynucleotide according to claim 1 , 2 or 3 encoding a protein that activates NF-κB, and a component that provides a detectable signal associated with activation of NF-κB;
(b) culturing a transformed cell under conditions, which permit the expression of the gene in the transformed cell;
(c) contacting the transformed cell with one or more compounds;
(d) measuring the detectable signal; and
(e) isolating or identifying an activator compound and/or an inhibitor compound by measuring the detectable signal.
10. A recombinant vector which comprises a polynucleotide according to claim 5 .
11. A transformed cell which comprises the recombinant vector according to claim 10 .