Nucleotide sequence encoding a modulator of nf-kb

Abstract

The present invention relates to nucleotide sequences encoding a modulator of NF-κB, and to the polypeptides encoded by the nucleotide sequences. In particular, the invention relates to nucleotide sequences and the polypeptides encoded thereby, wherein the polypeptides are involved in the response to NF-κB-activating stimuli, including HTLV-1 Tax, LPS, PMA and IL-1. The invention also relates to antibodies to the modulator of NF-κB, methods of detecting modulator of NF-κB using the antibodies, methods of treatment associated with NF-κB activation and to methods of identifying compounds which modulate the activity of the modulator of NF-κB.

1 - 11 . (canceled) 
     
     
         12 . A purified DNA sequence which encodes a purified modulator of NF-κB, or a fragment thereof which binds to a component of an IκB kinase, selected from the group consisting of:
 (A) the DNA sequence of SEQ ID NO: 2;   (B) DNA sequences that hybridize to the DNA sequence of (A) under standard hybridization conditions; and   (C) DNA sequences that encode a purified modulator of NF-κB, comprising a material selected from the group consisting of a protein, active fragments thereof, agonists thereof, mimics thereof and combinations thereof, said modulator having the following characteristics:
 a) an apparent molecular weight of approximately 48 kD; 
 b) a pI of approximately 5.66: 
 c) containing a leucine zipper motif; and 
 d) binding to a kinase involved in the activation of NF-κB. 
   
     
     
         13 . A recombinant DNA molecule comprising the purified DNA sequence of  claim 12 . 
     
     
         14 . The recombinant DNA molecule of  claim 13 , wherein said DNA sequence is operatively linked to an expression control sequence. 
     
     
         15 . The recombinant DNA molecule of  claim 14 , wherein said expression control sequence is selected from the group consisting of the early or late promoters of CMV, the lac system, the trp system, the TAC system, the TRC system, the major operator and promoter regions of phage λ, the control regions of fd coat protein, the promoter for 3-phosphoglycerate kinase, the promoters of acid phosphatase and the promoters of the yeast α-mating factors. 
     
     
         16 . A probe capable of screening for a DNA sequence encoding a modulator of NF-κB in alternate species, wherein said probe is a fragment of the purified DNA sequence of  claim 12  which binds to the DNA sequence encoding a modulator of NF-κB in alternate species under standard hybridization conditions. 
     
     
         17 . A unicellular host transformed with the recombinant DNA molecule of  claim 14 . 
     
     
         18 . The unicellular host of  claim 17  wherein the unicellular host is selected from the group consisting of  E. coli, Pseudomonas, Bacillus, Streptomyces , yeasts, CHO, R1.1, B-W, L-M, COS 1, COS 7, BSC1, BSC40, and BMT10 cells, plant cells, insect cells, and human cells in tissue culture. 
     
     
         19 . A method for producing a purified modulator of NF-κB, comprising a material selected from the group consisting of a protein, active fragments thereof, agonists thereof, mimics thereof, and combinations thereof, said modulator having the following characteristics:
 a) an apparent molecular weight of approximately 48 kD;   b) a pI of approximately 5.66;   c) containing a leucine zipper motif; and   d) binding to a kinase involved in the activation of NF-κB,   
       wherein said method comprises:
 (a) culturing the unicellular host of  claim 17 ; and 
 (b) isolating the modulator of NF-κB from said cultured unicellular host. 
 
     
     
         20 - 30 . (canceled) 
     
     
         31 . A recombinant virus transformed with the DNA molecule of  claim 12 . 
     
     
         32 - 33 . (canceled) 
     
     
         34 . An antisense nucleic acid against an mRNA encoding a purified modulator of NF-κB, comprising a material selected from the group consisting of a protein, active fragments thereof, agonists thereof, mimics thereof, and combinations thereof, said modulator having the following characteristics:
 a) an apparent molecular weight of approximately 48 kD;   b) a pI of approximately 5.66;   c) containing a leucine zipper motif; and   d) binding to a kinase involved in the activation of NF-κB,   
       wherein said antisense nucleic acid comprises a nucleic acid sequence hybridizing to said mRNA. 
     
     
         35 . The antisense nucleic acid of  claim 34  which is RNA. 
     
     
         36 . The antisense nucleic acid of  claim 35  which is DNA. 
     
     
         37 . The antisense nucleic acid of  claim 35  which binds to the initiation codon of said mRNA. 
     
     
         38 . A recombinant DNA molecule having a DNA sequence which, on transcription, produces an antisense ribonucleic acid against an mRNA encoding a purified modulator of NF-κB, comprising a material selected from the group consisting of a protein, active fragments thereof, agonists thereof, mimics thereof, and combinations thereof, said modulator having the following characteristics:
 a) an apparent molecular weight of approximately 48 kD;   b) a pI of approximately 5.66;   c) containing a leucine zipper motif; and   d) binding to a kinase involved in the activation of NF-κB,   wherein said antisense ribonucleic acid comprising an nucleic acid sequence capable of hybridizing to said mRNA.   
     
     
         39 . A cell line transfected with the recombinant DNA molecule of  claim 38 . 
     
     
         40 . A method for creating a cell line which exhibits reduced transcriptional activity, comprising transfecting an NF-κB producing cell line with the recombinant DNA molecule of  claim 38 . 
     
     
         41 - 50 . (canceled)