DNA-damage-induced proteolysis
The present application provides methods for identifying compounds for inhibiting DNA damage-induced Htt proteolysis, and methods and compositions for protecting cells from DNA damage-induced cleavage of Htt.
1. A method of identifying a compound for protecting a cell from DNA damage-induced Htt proteolysis, comprising:
providing one or more compounds to be tested;
identifying which compound is an IKKβ inhibitor;
testing the compound identified as the IKKβ inhibitor for its ability to reduce cleavage of Htt in a cell.
2. The method of claim 1 , wherein the cell is a neuron.
3. The method of claim 1 , wherein the Htt is wild type Htt.
4. The method of claim 1 , wherein the Htt is mutant Htt.
5. The method of claim 1 , wherein the ability of the compound to reduce the cleavage of Htt is tested by monitoring a cell's survival.
6. The method of claim 1 , wherein the compound is selected from the group consisting of a small molecule, a nucleic acid, a peptide, and an antibody.
7. The method of claim 1 , wherein identifying the IKKβ inhibitor comprises testing a compound for its ability to inhibit the ability of IKKβ to phosphorylate a substrate.
8. The method of claim 7 , wherein the substrate is Bcl-xL.
9. The method of claim 7 , wherein the compound is tested for its ability to inhibit IKKβ phosphorylation in an immune kinase assay.
10. The method of claim 1 , wherein identifying the IKKβ inhibitor comprises testing a compound for its ability to inhibit activation of one or more caspases.
11. The method of claim 2 , wherein the one or more caspases are selected from the group consisting of caspase-3 and caspase-6.
12. The method of claim 1 , further comprising the step of applying a DNA damaging agent to the cell.
13. A method of identifying a compound for protecting a cell from DNA damage-induced Htt proteolysis, comprising:
providing one or more compound to be tested;
identifying which compound is selected from the group consisting of IKKα activator, Bcl-xL inducer, and some combination thereof;
testing the compound identified as an IKKα activator, Bcl-xL inducer, or some combination thereof for its ability to reduce the cleavage of Htt in a cell.
14. The method of claim 13 , wherein the cell is a neuron.
15. The method of claim 13 , wherein the Htt is wild type Htt.
16. The method of claim 13 , wherein the Htt is mutant Htt.
17. The method of claim 13 , wherein the compound is selected from the group consisting of a small molecule, a nucleic acid, a peptide, and an antibody.
18. The method of claim 13 , wherein the ability of the compound to reduce the cleavage of Htt is tested by monitoring a cell's survival.
19. The method of claim 13 , wherein the ability of the compound to reduce the cleavage of Htt is assayed through monitoring the cleaved Htt product.
20. The method of claim 13 , wherein identifying the IKKα activator comprises testing the compound for its ability to inhibit the phosphorylation of Bcl-xL.
21. The method of claim 13 , wherein identifying the IKKα activator comprises testing the compound for its ability to inhibit activation of one or more caspases.
22. The method of claim 21 , wherein the one or more caspases are selected from the group consisting of caspase-3 and caspase-6.
23. The method of claim 13 , wherein identifying the Bcl-xL inducer comprises testing compounds for its ability to inhibit phosphorylation of Bcl-xL.
24. The method of claim 1 , further comprising the step of applying a DNA damaging agent to the cell.