Nucleotide sequence encoding a modulator of nf-kb
a technology of nf-kb and nucleotide sequence, applied in the field of polypeptides, can solve the problem of reducing the resulting kinase activity, and achieve the effect of enhancing such activity
Inactive Publication Date: 2009-03-05
INST PASTEUR
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- Description
- Claims
- Application Information
AI Technical Summary
Benefits of technology
The invention provides a way to purify a protein called the modulator of NF-κB and its subunits, which can inhibit the activity of NF-κB. The invention also provides antibodies to the modulator and methods for detecting its presence in mammals. The invention also describes methods for screening substances that can mimic or fight against the modulator, as well as treating mammals to control the transcriptional activity of NF-κB. The invention also includes pharmaceutical compositions for treating diseases associated with the modulator.
Problems solved by technology
Another important issue unanswered was how discrete activation signals triggered by a variety of known stimulators are integrated to give rise to IκB kinase activity.
Deletion of the HLH motif leaves the coimmunoprecipitation of the two kinases intact, but strongly reduces the resulting kinase activity.
Method used
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example 1
Materials and Methods
[0154]Cells and transfections. The 70Z / 3 murine pre-B cell line and the NF-1 unresponsive mutant 1.3E2 were maintained in RPMI medium supplemented with 10% foetal calf serum and 50 μM β-mercaptoethanol. 70Z / 3 and 1.3E2 cells were transiently transfected as described (Courtois et al., 1997). Isolated stable clones were prepared as described (Whiteside et al., 1995). Rat-1 and 5R cells were grown in DMEM supplemented with 10% foetal calf serum and transfected using the calcium phosphate coprecipitation method. For measurement of luciferase activity in transiently transfected Rat-1 or 5R cells, approximately 2×105 cells were transfected with 0.25 μg of a reporter plasmid, 0.25 μg of EF1-lacZ plasmid and 1 μg of either vector or effector plasmid. Cells were harvested 40 to 45 hours after transfection. The amount of lysate used for luciferase assay was determined on the basis of β-galactosidase activity. The results shown are representative of one experiment carried ...
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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.
Description
BACKGROUND OF THE INVENTION[0001]1. Field of the Invention[0002]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.[0003]2. Discussion of the Background[0004]The Rel / NF-κB family of transcription factors plays important roles in immune and stress responses, in inflammation, in apoptosis, and regulates the expression of numerous cellular and viral genes (for recent reviews, see Baldwin, 1996; Verma et al., 1995; May and Ghosh, 1998). The NF-κB activity is composed of homo- or heterodimers of related proteins that share a conserved DNA-binding and dimerization domain called the Rel Homology Domain. In most cell types, NF-κB is sequestered in the cytoplasm bound to inhibitor...
Claims
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IPC IPC(8): C12P21/06C07H21/04C12N15/63C12N1/21C12N1/19C07H21/02C12N7/01C12N5/16C12N5/14C12N5/22A61K38/00C07K14/47C07K16/18G01N33/68
CPCG01N2500/04G01N2500/10A61K38/00G01N2500/02C07K16/18G01N33/6872C07K14/4702A61P29/00A61P37/00A61P43/00Y02A50/30
InventorYAMAOKA, SHOJICOURTOIS, GILLESISRAEL, ALAINWEIL, ROBERT
OwnerINST PASTEUR