Huntington's disease: from experimental results to interaction networks, patho-pathway construction and disease hypothesis
© Gonzalez-Couto et al; licensee BioMed Central Ltd. 2007
Published: 8 May 2007
Protein-protein interaction networks and mechanistic pathway models are excellent tools in the drug discovery process. They can be used to identify and select targets for a given disease hypothesis. Combining information from diverse sources, like in house experiments as well as literature, allows further development of interaction networks into detailed descriptions of cellular pathways. Computerized pathway diagrams allow integrating all relevant data regarding a project into one framework by linking the different data sources. Interaction networks analysis and pathway design tools are used to support target identification and validation activities. Experimental results are incorporated in protein interaction networks, analyzed and further developed into biomolecular pathopathways including literature findings to understand the underlying modulation mechanisms. The pathway diagrams are also used as communication tools, particularly for interdisciplinary project teams, thus ensuring a common understanding and facilitating critical interrogations about disease hypotheses. The analyses of experimental results, the initial construction of an HD pathopathway are presented and two mechanistic disease hypotheses are discussed.
Materials and methods
Differential proteomics experiments (DPEs) were performed on rat PC12 cells containing either wild-type or mutant full-length (PolyQ) Huntingtin (Htt) under control of a doxycycline-inducible promoter . Cell extracts were prepared at 0, 12, and 48 hours post-induction to identify proteins involved in pre-apoptotic intracellular events. Protein expression modulation was measured using DIGE technology  followed by statistical analysis for spot selection and automated spot picking. The protein content of each picked spot was analyzed by mass spectrometry (MS).
DPE results were used to develop an HD pathopathway including proteins modulated by PolyQ Htt expression. In this experimental result set, the proteins from the Ubiquitin-Proteasome pathway were chosen for in depth analysis, and UCHL1 was identified as a component potentially playing opposite roles in HD.
- Sugars KL, Brown R, Cook LJ, Swartz J, Rubinsztein DC: Decreased cAMP response element-mediated transcription: an early event in exon 1 and full-length cell models of Huntington's disease that contributes to polyglutamine pathogenesis. J Biol Chem. 2004, 279: 4988-4999. 10.1074/jbc.M310226200PubMedView ArticleGoogle Scholar
- Raggiaschi R, Gotta S, Terstappen GC: Phosphoproteome analysis. Biosci Rep. 2005, 25: 33-44. 10.1007/s10540-005-2846-0PubMedView ArticleGoogle Scholar
- Gygi SP, Corthals GL, Zhang Y, Rochon Y, Aebersold R: Evaluation of two-dimensional gel electrophoresis-based proteome analysis technology. Proc Natl Acad Sci USA. 2000, 97: 9390-9395. 10.1073/pnas.160270797PubMedPubMed CentralView ArticleGoogle Scholar
- Hol EM, Fischer DF, Ovaa H, Scheper W: Ubiquitin proteasome system as a pharmacological target in neurodegeneration. Expert Rev Neurother. 2006, 6: 1337-1347. 10.1586/1473722.214.171.1247PubMedView ArticleGoogle Scholar
- Liu Y, Fallon L, Lashuel HA, Liu Z, Lansbury PT: The UCHL1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinson's disease susceptibility. Cell. 2002, 111: 209-218. 10.1016/S0092-8674(02)01012-7PubMedView ArticleGoogle Scholar
- Gong B, Cao Z, Zheng P, Vitolo OV, Liu S, Staniszewski A, Moolman D, Zhang H, Shelanski M, Arancio O: Ubiquitin hydrolase UCHL1 rescues beta-amyloid-induced decreases in synaptic function and contextual memory. Cell. 2006, 126: 775-788. 10.1016/j.cell.2006.06.046PubMedView ArticleGoogle Scholar
- Naze P, Vuillaume I, Destee A, Pasquier F, Sablonniere B: Mutation analysis and association studies of the ubiquitin carboxy-terminal hydrolase L1 gene in Huntington's disease. Neurosci Lett. 2002, 328: 1-4. 10.1016/S0304-3940(02)00231-8PubMedView ArticleGoogle Scholar
- Metzger S, Bauer P, Tomiuk J, Laccone F, Didonato S, Gellera C, Soliveri P, Lange HW, Weirich-Schwaiger H, Wenning GK: The S18Y polymorphism in the UCHL1 gene is a genetic modifier in Huntington's disease. Neurogenetics. 2006, 7: 27-30. 10.1007/s10048-005-0023-zPubMedView ArticleGoogle Scholar
- Galvao R, Mendes-Soares L, Camara J, Jaco I, Carmo-Fonseca M: Triplet repeats, RNA secondary structure and toxic gain-of-function models for pathogenesis. Brain Res Bull. 2001, 56: 191-201. 10.1016/S0361-9230(01)00651-7PubMedView ArticleGoogle Scholar
- Duennwald ML, Jagadish S, Giorgini F, Muchowski PJ, Lindquist S: A network of protein interactions determines polyglutamine toxicity. Proc Natl Acad Sci USA. 2006, 103: 11051-11056. 10.1073/pnas.0604548103PubMedPubMed CentralView ArticleGoogle Scholar
- Nicastro G, Menon RP, Masino L, Knowles PP, McDonald NQ, Pastore A: The solution structure of the Josephin domain of ataxin-3: structural determinants for molecular recognition. Proc Natl Acad Sci USA. 2005, 102: 10493-10498. 10.1073/pnas.0501732102PubMedPubMed CentralView ArticleGoogle Scholar
- Menon RP, Pastore A: Expansion of amino acid homo-sequences in proteins: insights into the role of amino acid homo-polymers and of the protein context in aggregation. Cell Mol Life Sci. 2006, 63: 1677-1685. 10.1007/s00018-006-6097-4PubMedView ArticleGoogle Scholar
- Ellisdon AM, Thomas B, Bottomley SP: The two-stage pathway of ataxin-3 fibrillogenesis involves a polyglutamine-independent step. J Biol Chem. 2006, 281: 16888-16896. 10.1074/jbc.M601470200PubMedView ArticleGoogle Scholar
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