Compositions and methods for gene expression
View Patent ↗The invention provides nucleotide sequences that mediate one or more functions of IKKα, kits and methods for using these sequences to identify therapeutic compounds that alter IKKα related pathology.
1. A method for identifying one or more test compounds that alters binding of RelB Rel homology domain (RelB RHD) with RelBκB sequence, comprising:
a) providing
i) an isolated nucleotide sequence comprising 5′-NGGAGANNTG-3′ (SEQ ID NO:57); wherein N at position 1 is chosen from G and A, N at position 7 is chosen from T and C, and N at position 8 is chosen from T and C, and wherein said isolated sequence specifically binds with a polypeptide sequence comprising RelB Rel homology domain (RHD) as comprising SEQ ID NO:62,
ii) a polypeptide comprising RelB RHD comprising SEQ ID NO:62, and
iii) one or more test compounds;
b) contacting said isolated nucleotide sequence with said polypeptide in the presence and absence of said one or more test compounds; and
determining the level of specific binding of said nucleotide sequence with said polypeptide in the presence of said one or more test compounds compared to in the absence of said one or more test compounds, wherein detecting altered specific binding of said nucleotide sequence with said polypeptide in the presence of said one or more test compounds compared to in the absence of said one or more test compounds identifies said one or more test compounds as altering binding of RelB RHD with RelBκB sequence.
2. The method claim 1 , wherein said polypeptide is recombinant.
3. The method of claim 2 , wherein said polypeptide comprises RelB:p52.
4. The method of claim 2 , wherein said polypeptide comprises RelB.
5. The method of claim 2 , wherein said contacting is in vivo.
6. The method of claim 2 , wherein said contacting is in vitro.
7. The method of claim 1 , further comprising detecting unaltered binding of said nucleotide sequence to a protein comprising one or more of (a) RelA Rel homology domain (RelA RHD) comprising SEQ ID NO:65, (b) RelA, (c) p50, (d) RelA:p50, (e) p52, and(f) RelA:p52, in the presence and absence of said one or more test compounds, wherein said unaltered binding indicates specific binding of said nucleotide sequence with said polypeptide.
8. The method of claim 1 , further comprising detecting unaltered binding of an isolated nucleotide sequence comprising the consensus-KB sequence 5′-GGGACTTTCC-3′ (SEQ ID NO:58) to a polypeptide comprising one or more of RelB RHD comprising SEQ ID NO:62, and RelB in the presence of said one or more test compounds, wherein said unaltered binding indicates specific binding of said nucleotide sequence with said polypeptide.