The Journal of Neuroscience, December 14, 2005, ():
Dysregulation of Receptor Interacting Protein-2 and Caspase Recruitment Domain Only Protein Mediates Aberrant Caspase-1 Activation in Huntington's Disease
J. Neurosci. Wang et al.
25: 11645
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplementary Figure S1. HeLa cells and mutant-htt ST14A cells were transfected with a FLAG-tagged Cop construct. HeLa cells were either IP (line 3) with a FLAG antibody or a normal mouse IgG (as a control, line 1) and then immunblotted with a Cop (1:100) antibody, or directly analyzed (lysate, line 2) by western blotting with a Cop (1:100) antibody (A), or submitted to immunofluorescent staining with Cop (1:100) antibodies and Texas Red-conjugated secondary antibodies (B). These results confirm that this antibody can recognize Cop protein. H. C., Heavy Chain. L. C., Light Chain. Exogenous Cop in mutant-htt ST14A was detected by either FLAG antibodies and FITC-conjugated secondary antibodies (1:300) (C) or Cop (1:100) antibodies and Texas Red-conjugated secondary antibodies (1:300) (D, E).
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Supplementary Fig. S 2. Dysregulation of Rip2 and Cop results in aberrant caspase-1 activation, and plays a key role in mutant huntingtin-mediated neurodegeneration in Huntington's disease. The central portion of this diagram recapitulates the above-described data on Rip2 and Cop protein and their antagonistic effects on the activation of caspase-1. In normal conditions, wild type huntingtin (WT htt) does not cause upregulation of pro-caspase-1, there are high levels of Cop, and low levels of Rip2. Pro-caspase-1 remains in its inactive form. In Huntington’s disease, mutant huntingtin mediates upregulation of caspase-1 mRNA and protein. In addition, therefore mutant huntingtin expression results in lowering of Cop levels and increasing of Rip2 levels, thereafter mediating caspase-1 activation. Caspase-1 thereafter mediates the activation of downstream cell death pathways. The mechanisms by which mutant huntingtin mediates the downregulation of Cop and the upregulation of Rip2 are at present not understood, and a subject of current investigation.
