The yeast two-hybrid (Y2H) system is the most widely applied methodology for systematic protein-protein interaction (PPI) screening and the generation of comprehensive interaction networks. We developed a novel Y2H interaction screening procedure using DNA microarrays for high-throughput quantitative PPI detection. Applying a global pooling and selection scheme to a large collection of human open reading frames, proof-of-principle Y2H interaction screens were performed for the human neurodegenerative disease proteins huntingtin and ataxin-1. Using systematic controls for unspecific Y2H results and quantitative benchmarking, we identified and scored a large number of known and novel partner proteins for both huntingtin and ataxin-1. Moreover, we show that this parallelized screening procedure and the global inspection of Y2H interaction data are uniquely suited to define specific PPI patterns and their alteration by disease-causing mutations in huntingtin and ataxin-1. This approach takes advantage of the specificity and flexibility of DNA microarrays and of the existence of solid-related statistical methods for the analysis of DNA microarray data, and allows a quantitative approach toward interaction screens in human and in model organisms.
OPTN, STAM2, EIF1B, HMG20A, BASP1, UBAC1, TRIM38, UBE2E3, RAI2, ARID5A, TRAFD1, IMMT, ILVBL, DUSP10, CHRNA7, MRFAP1L1, UHRF2, FAM46B, CLCN2, ADD3, CREM, CRIP2, CRK, CRY2, ZMAT2, MAGEB6, XAGE3, ZFC3H1, DHRSX, ESRRA, ETV4, ATF5, GGA2, UHRF1BP1L, ZDHHC17, HEY2, RBFOX2, FTL, HYPM, ZNF451, HEYL, OSTF1, NPHP3, ZSCAN1, MAGEB18, RAPGEF1, CDIP1, HTT, UBE2K, HMGA1, HOXC4, HOXC11, IGF2R, KIF22, LASP1, LOXL1, C2orf27B, MAGEB2, MBD1, MIPEP, ASNS, MSX2, NCAM1, NME4, OPCML, OTX2, TMX2, EVL, TDP2, PIK3R1, PPAT, ERCC6L, GOLPH3L, DPPA4, FAM46A, RNF20, PSMB7, PSPH, DHX37, SEMA4G, GATAD1, PLEKHB1, RPL4, ATXN1, RBM26, MLST8, TTC23, UPF3A, SLC4A2, SLC6A13, TACC1, TCTA, TSC1, YWHAE, SF1, ZNF655, KCTD15, ZXDC, EHMT1, BAALC, DOCK5, PTGES2, FOSL1, RCC1L, CAMK2B, HIST1H3A, HIST1H3D, HIST1H3C, HIST1H3E, HIST1H3I, HIST1H3G, HIST1H3J, HIST1H3H, HIST1H3B, FAR1, ING5, FAM71F1, ZC3H10, ZFYVE19, DIXDC1, PIAS1, EIF3F, PEX11B, MBD4, FUBP3, HIST1H3F, IFT20, CCDC126, LPAR2, SLC25A51, HNRNPLL, MRFAP1, MED21, QKI, LITAF, TP53I11, TOMM20, NUP58, SV2A