IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ DJ-1 binds androgen receptor directly and mediates its activity in hormonally treated prostate cancer cells. DJ-1 binds androgen receptor directly and mediates its activity in hormonally treated prostate cancer cells. Cancer Res. (0008-5472) 2007 imex curation Tillman JE., Yuan J., Gu G., Fazli L., Ghosh R., Flynt AS., Gleave M., Rennie PS., Kasper S. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Cancer - Interactions investigated in the context of cancer I calculated the range of aa based on the range given in bp. susan.kasper@vanderbilt.edu A cDNA library generated from flutamide-activated primary HPE cells was used. orchard Only protein-protein interactions human-lncap Human caucasian prostate carcinoma lncap Human caucasian prostate adenocarcinoma confocal microscopy confocal microscopy predetermined predetermined participant predetermined Tillman JE., Yuan J., Gu G., Fazli L., Ghosh R., Flynt AS., Gleave M., Rennie PS., Kasper S. Parkinsons - Interactions investigated in the context of Parkinsons disease susan.kasper@vanderbilt.edu Accepted 2007-NOV-17 by ORCHARD. Cancer Res. (0008-5472) 2007 orchard Only protein-protein interactions imex curation DJ-1 binds androgen receptor directly and mediates its activity in hormonally treated prostate cancer cells. DJ-1 binds androgen receptor directly and mediates its activity in hormonally treated prostate cancer cells. Cancer Res. (0008-5472) 2007 imex curation Tillman JE., Yuan J., Gu G., Fazli L., Ghosh R., Flynt AS., Gleave M., Rennie PS., Kasper S. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Cancer - Interactions investigated in the context of cancer I calculated the range of aa based on the range given in bp. susan.kasper@vanderbilt.edu A cDNA library generated from flutamide-activated primary HPE cells was used. orchard Only protein-protein interactions human-lncap Human caucasian prostate carcinoma lncap Human caucasian prostate adenocarcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Tillman JE., Yuan J., Gu G., Fazli L., Ghosh R., Flynt AS., Gleave M., Rennie PS., Kasper S. Parkinsons - Interactions investigated in the context of Parkinsons disease susan.kasper@vanderbilt.edu Accepted 2007-NOV-17 by ORCHARD. Cancer Res. (0008-5472) 2007 orchard Only protein-protein interactions imex curation DJ-1 binds androgen receptor directly and mediates its activity in hormonally treated prostate cancer cells. DJ-1 binds androgen receptor directly and mediates its activity in hormonally treated prostate cancer cells. Cancer Res. (0008-5472) 2007 imex curation Tillman JE., Yuan J., Gu G., Fazli L., Ghosh R., Flynt AS., Gleave M., Rennie PS., Kasper S. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Cancer - Interactions investigated in the context of cancer I calculated the range of aa based on the range given in bp. susan.kasper@vanderbilt.edu A cDNA library generated from flutamide-activated primary HPE cells was used. orchard Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Tillman JE., Yuan J., Gu G., Fazli L., Ghosh R., Flynt AS., Gleave M., Rennie PS., Kasper S. Parkinsons - Interactions investigated in the context of Parkinsons disease susan.kasper@vanderbilt.edu Accepted 2007-DEC-1 by ORCHARD. Cancer Res. (0008-5472) 2007 orchard Only protein-protein interactions imex curation DJ-1 binds androgen receptor directly and mediates its activity in hormonally treated prostate cancer cells. DJ-1 binds androgen receptor directly and mediates its activity in hormonally treated prostate cancer cells. Cancer Res. (0008-5472) 2007 imex curation Tillman JE., Yuan J., Gu G., Fazli L., Ghosh R., Flynt AS., Gleave M., Rennie PS., Kasper S. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Cancer - Interactions investigated in the context of cancer I calculated the range of aa based on the range given in bp. susan.kasper@vanderbilt.edu A cDNA library generated from flutamide-activated primary HPE cells was used. orchard Only protein-protein interactions human-lapc4 Homo sapiens LAPC4 cells derived from a human prostate cancer xenograft lapc4 Human prostate cancer xenograft cell line LAPC4 LAPC4 cells were derived from a human prostate cancer xenograft implanted in SCID mice and express wild-type androgen receptor anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Tillman JE., Yuan J., Gu G., Fazli L., Ghosh R., Flynt AS., Gleave M., Rennie PS., Kasper S. Parkinsons - Interactions investigated in the context of Parkinsons disease susan.kasper@vanderbilt.edu Accepted 2007-NOV-17 by ORCHARD. Cancer Res. (0008-5472) 2007 orchard Only protein-protein interactions imex curation DJ-1 binds androgen receptor directly and mediates its activity in hormonally treated prostate cancer cells. DJ-1 binds androgen receptor directly and mediates its activity in hormonally treated prostate cancer cells. Cancer Res. (0008-5472) 2007 imex curation Tillman JE., Yuan J., Gu G., Fazli L., Ghosh R., Flynt AS., Gleave M., Rennie PS., Kasper S. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Cancer - Interactions investigated in the context of cancer I calculated the range of aa based on the range given in bp. susan.kasper@vanderbilt.edu A cDNA library generated from flutamide-activated primary HPE cells was used. orchard Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Tillman JE., Yuan J., Gu G., Fazli L., Ghosh R., Flynt AS., Gleave M., Rennie PS., Kasper S. Parkinsons - Interactions investigated in the context of Parkinsons disease I calculated the range of aa based on the range given in bp. susan.kasper@vanderbilt.edu Accepted 2007-DEC-1 by ORCHARD. A cDNA library generated from flutamide-activated primary HPE cells was used. Cancer Res. (0008-5472) 2007 orchard Only protein-protein interactions imex curation DJ-1 binds androgen receptor directly and mediates its activity in hormonally treated prostate cancer cells. DJ-1 binds androgen receptor directly and mediates its activity in hormonally treated prostate cancer cells. Cancer Res. (0008-5472) 2007 imex curation Tillman JE., Yuan J., Gu G., Fazli L., Ghosh R., Flynt AS., Gleave M., Rennie PS., Kasper S. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Cancer - Interactions investigated in the context of cancer I calculated the range of aa based on the range given in bp. susan.kasper@vanderbilt.edu A cDNA library generated from flutamide-activated primary HPE cells was used. orchard Only protein-protein interactions human-lapc4 Homo sapiens LAPC4 cells derived from a human prostate cancer xenograft lapc4 Human prostate cancer xenograft cell line LAPC4 LAPC4 cells were derived from a human prostate cancer xenograft implanted in SCID mice and express wild-type androgen receptor anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Tillman JE., Yuan J., Gu G., Fazli L., Ghosh R., Flynt AS., Gleave M., Rennie PS., Kasper S. Parkinsons - Interactions investigated in the context of Parkinsons disease susan.kasper@vanderbilt.edu Accepted 2007-NOV-17 by ORCHARD. Cancer Res. (0008-5472) 2007 orchard Only protein-protein interactions imex curation park7_human Parkinson disease protein 7 PARK7 Oncogene DJ1 Parkinsonism-associated deglycase Maillard deglycase Protein DJ-1 Protein/nucleic acid deglycase DJ-1 protein protein human Homo sapiens Human MASKRALVILAKGAEEMETVIPVDVMRRAGIKVTVAGLAGKDPVQCSRDVVICPDASLEDAKKEGPYDVVVLPGGNLGAQNLSESAAVKEILKEQENRKGLIAAICAGPTALLAHEIGFGSKVTTHPLAKDKMMNGGHYTYSENRVEKDGLILTSRGPGTSFEFALAIVEALNGKEVAAQVKAPLVLKD DIP protein Q99497 original sequence version: 102 4B21661B3A76BC67 andr_human Androgen receptor Dihydrotestosterone receptor Nuclear receptor subfamily 3 group C member 4 AR DHTR NR3C4 protein protein human Homo sapiens Human MEVQLGLGRVYPRPPSKTYRGAFQNLFQSVREVIQNPGPRHPEAASAAPPGASLLLLQQQQQQQQQQQQQQQQQQQQQQQETSPRQQQQQQGEDGSPQAHRRGPTGYLVLDEEQQPSQPQSALECHPERGCVPEPGAAVAASKGLPQQLPAPPDEDDSAAPSTLSLLGPTFPGLSSCSADLKDILSEASTMQLLQQQQQEAVSEGSSSGRAREASGAPTSSKDNYLGGTSTISDNAKELCKAVSVSMGLGVEALEHLSPGEQLRGDCMYAPLLGVPPAVRPTPCAPLAECKGSLLDDSAGKSTEDTAEYSPFKGGYTKGLEGESLGCSGSAAAGSSGTLELPSTLSLYKSGALDEAAAYQSRDYYNFPLALAGPPPPPPPPHPHARIKLENPLDYGSAWAAAAAQCRYGDLASLHGAGAAGPGSGSPSAAASSSWHTLFTAEEGQLYGPCGGGGGGGGGGGGGGGGGGGGGGGEAGAVAPYGYTRPPQGLAGQESDFTAPDVWYPGGMVSRVPYPSPTCVKSEMGPWMDSYSGPYGDMRLETARDHVLPIDYYFPPQKTCLICGDEASGCHYGALTCGSCKVFFKRAAEGKQKYLCASRNDCTIDKFRRKNCPSCRLRKCYEAGMTLGARKLKKLGNLKLQEEGEASSTTSPTEETTQKLTVSHIEGYECQPIFLNVLEAIEPGVVCAGHDNNQPDSFAALLSSLNELGERQLVHVVKWAKALPGFRNLHVDDQMAVIQYSWMGLMVFAMGWRSFTNVNSRMLYFAPDLVFNEYRMHKSRMYSQCVRMRHLSQEFGWLQITPQEFLCMKALLLFSIIPVDGLKNQKFFDELRMNYIKELDRIIACKRKNPTSCSRRFYQLTKLLDSVQPIARELHQFTFDLLIKSHMVSVDFPEMMAEIISVQVPKILSGKVKPIYFHTQ DIP protein P10275 original sequence version: 220 DIP protein P10275-1 original sequence version: 213 DIP protein P10275 original sequence version: 216 DIP protein P10275 original sequence version: 143 DIP protein P10275 original sequence version: 174 DIP protein P10275 original sequence version: 169 DIP protein P10275 original sequence version: 188 DIP protein P10275-1 original sequence version: 188 DIP protein P10275 original sequence version: 196 DIP protein P10275 original sequence version: 217 DIP protein P10275-1 original sequence version: 217 DIP protein P10275-1 original sequence version: 207 A73432C55D39AE06 park7-ar 5 7 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Full-length DJ-1 cDNA was used to screen the library and identified a region of AR (Fig. 1B). The AR fragment contained a region of the NH2-terminal activation domain, the entire DNA binding domain, and a portion of the ligand binding domain park7-ar-1 3 8 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role bait bait physical association physical association high-stringency selection showed that neither the bait (DJ-1) nor prey (AR) plasmid was capable of activating reporter genes individually,but the combination produced an interaction and activation of the reporters. 1 B ar-park7 4 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association 1 C Immunoprecipitation of endogenous AR pulled down DJ-1 Dihydrotestosterone (DHT) - Treatment with DHT increased the interaction between DJ-1 and AR. park7-ar-2 6 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 1 D The reciprocal immunoprecipitation was done in LAPC4cells expressing a NH2-terminally HA-tagged DJ-1. Immunoprecipitation with anti-HA antibody coimmunoprecipitated HA-tagged DJ-1, endogenous DJ-1, and AR. DJ-1 exists as an obligatory dimer Q99497 transfection ar-park7-1 2 8 cofactor cofactor coenzyme bait bait 7 cofactor cofactor coenzyme prey prey physical association physical association 2 Dihydrotestosterone (DHT) - Treatment with DHT increased the interaction between DJ-1 and AR. In both LAPC4and LNCaP cells, antiandrogen treatment with either OH-flutamide or bicalutamide increased the amount of DJ-1 bound to AR ar-park7-2 1 7 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization 3 Yellow pixels from the LNCaP images in Fig. 3C were displayed alone and indicate that DJ-1-AR colocalization increased during hormonal treatment DJ-1-AR colocalization was greatest with DHT treatment, followed by OH-flutamide and bicalutamide. DHT and bicalutamide treatment resulted in similar colocalization throughout the nucleus, whereas DJ-1-AR with OH-flutamide treatment seemed to concentrate just within the nuclear perimeter.