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/ http://www.ebi.ac.uk/ wafa-2003-1 Isolation and identification of L-dopa decarboxylase as a protein that binds to and enhances transcriptional activity of the androgen receptor using the repressed transactivator yeast two-hybrid system. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast reverse two hybrid reverse two hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Wafa LA., Cheng H., Rao MA., Nelson CC., Cox M., Hirst M., Sadowski I., Rennie PS. Cancer - Interactions investigated in the context of cancer A reverse yeast two-hybrid screen specifically designed for transactivator bait proteins was used. The AR NTD is fused to a GAL4 DBD to form the bait. The prey is created by fusion of the cDNA library with a TUP1 repressor protein. The N-terminus of TUP1 is sufficient to cause transcriptional repression, so only the first 200 amino acid residues were fused to the cDNA library. When there is no interaction between the bait and TUP1-library fusion protein, the bait activator allows constitutive expression of the URA3 reporter gene. This permits growth in the absence of uracil, but causes sensitivity to 5-FOA, which is converted into a toxic 5-flourouracil product through the enzyme encoded by the URA3 gene. Thus transcriptional repression can be genetically monitored by using a counter-selectable URA3 reporter. Where there is an interaction between the AR NTD and a library clone, TUP1 will cause repression of URA3 expression, which can be detected by growth of yeast in the presence of 5- FOA. Therefore, viability of the MAV103 strain, or colony growth, on 5-FOA media represents a positive interaction between the AR NTD bait and prey. prennie@interchange.ubc.ca A cDNA library from the LNCaP human prostate carcinoma cell line was used. Biochem. J. (0264-6021) 2003 Only protein-protein interactions imex curation wafa-2003-2 Isolation and identification of L-dopa decarboxylase as a protein that binds to and enhances transcriptional activity of the androgen receptor using the repressed transactivator yeast two-hybrid system. in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Wafa LA., Cheng H., Rao MA., Nelson CC., Cox M., Hirst M., Sadowski I., Rennie PS. Cancer - Interactions investigated in the context of cancer A reverse yeast two-hybrid screen specifically designed for transactivator bait proteins was used. The AR NTD is fused to a GAL4 DBD to form the bait. The prey is created by fusion of the cDNA library with a TUP1 repressor protein. The N-terminus of TUP1 is sufficient to cause transcriptional repression, so only the first 200 amino acid residues were fused to the cDNA library. When there is no interaction between the bait and TUP1-library fusion protein, the bait activator allows constitutive expression of the URA3 reporter gene. This permits growth in the absence of uracil, but causes sensitivity to 5-FOA, which is converted into a toxic 5-flourouracil product through the enzyme encoded by the URA3 gene. Thus transcriptional repression can be genetically monitored by using a counter-selectable URA3 reporter. Where there is an interaction between the AR NTD and a library clone, TUP1 will cause repression of URA3 expression, which can be detected by growth of yeast in the presence of 5- FOA. Therefore, viability of the MAV103 strain, or colony growth, on 5-FOA media represents a positive interaction between the AR NTD bait and prey. prennie@interchange.ubc.ca A cDNA library from the LNCaP human prostate carcinoma cell line was used. Biochem. J. (0264-6021) 2003 Only protein-protein interactions imex curation wafa-2003-3 Isolation and identification of L-dopa decarboxylase as a protein that binds to and enhances transcriptional activity of the androgen receptor using the repressed transactivator yeast two-hybrid system. 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 Wafa LA., Cheng H., Rao MA., Nelson CC., Cox M., Hirst M., Sadowski I., Rennie PS. Cancer - Interactions investigated in the context of cancer A reverse yeast two-hybrid screen specifically designed for transactivator bait proteins was used. The AR NTD is fused to a GAL4 DBD to form the bait. The prey is created by fusion of the cDNA library with a TUP1 repressor protein. The N-terminus of TUP1 is sufficient to cause transcriptional repression, so only the first 200 amino acid residues were fused to the cDNA library. When there is no interaction between the bait and TUP1-library fusion protein, the bait activator allows constitutive expression of the URA3 reporter gene. This permits growth in the absence of uracil, but causes sensitivity to 5-FOA, which is converted into a toxic 5-flourouracil product through the enzyme encoded by the URA3 gene. Thus transcriptional repression can be genetically monitored by using a counter-selectable URA3 reporter. Where there is an interaction between the AR NTD and a library clone, TUP1 will cause repression of URA3 expression, which can be detected by growth of yeast in the presence of 5- FOA. Therefore, viability of the MAV103 strain, or colony growth, on 5-FOA media represents a positive interaction between the AR NTD bait and prey. prennie@interchange.ubc.ca A cDNA library from the LNCaP human prostate carcinoma cell line was used. Biochem. J. (0264-6021) 2003 Only protein-protein interactions imex curation ddc_human Aromatic-L-amino-acid decarboxylase DDC AADC DOPA decarboxylase protein protein human Homo sapiens Human MNASEFRRRGKEMVDYMANYMEGIEGRQVYPDVEPGYLRPLIPAAAPQEPDTFEDIINDVEKIIMPGVTHWHSPYFFAYFPTASSYPAMLADMLCGAIGCIGFSWAASPACTELETVMMDWLGKMLELPKAFLNEKAGEGGGVIQGSASEATLVALLAARTKVIHRLQAASPELTQAAIMEKLVAYSSDQAHSSVERAGLIGGVKLKAIPSDGNFAMRASALQEALERDKAAGLIPFFMVATLGTTTCCSFDNLLEVGPICNKEDIWLHVDAAYAGSAFICPEFRHLLNGVEFADSFNFNPHKWLLVNFDCSAMWVKKRTDLTGAFRLDPTYLKHSHQDSGLITDYRHWQIPLGRRFRSLKMWFVFRMYGVKGLQAYIRKHVQLSHEFESLVRQDPRFEICVEVILGLVCFRLKGSNKVNEALLQRINSAKKIHLVPCHLRDKFVLRFAICSRTVESAHVQRAWEHIKELAADVLRAERE andr_human Androgen receptor Dihydrotestosterone receptor Nuclear receptor subfamily 3 group C member 4 AR DHTR NR3C4 protein protein human Homo sapiens Human MEVQLGLGRVYPRPPSKTYRGAFQNLFQSVREVIQNPGPRHPEAASAAPPGASLLLLQQQQQQQQQQQQQQQQQQQQQQQETSPRQQQQQQGEDGSPQAHRRGPTGYLVLDEEQQPSQPQSALECHPERGCVPEPGAAVAASKGLPQQLPAPPDEDDSAAPSTLSLLGPTFPGLSSCSADLKDILSEASTMQLLQQQQQEAVSEGSSSGRAREASGAPTSSKDNYLGGTSTISDNAKELCKAVSVSMGLGVEALEHLSPGEQLRGDCMYAPLLGVPPAVRPTPCAPLAECKGSLLDDSAGKSTEDTAEYSPFKGGYTKGLEGESLGCSGSAAAGSSGTLELPSTLSLYKSGALDEAAAYQSRDYYNFPLALAGPPPPPPPPHPHARIKLENPLDYGSAWAAAAAQCRYGDLASLHGAGAAGPGSGSPSAAASSSWHTLFTAEEGQLYGPCGGGGGGGGGGGGGGGGGGGGGGGEAGAVAPYGYTRPPQGLAGQESDFTAPDVWYPGGMVSRVPYPSPTCVKSEMGPWMDSYSGPYGDMRLETARDHVLPIDYYFPPQKTCLICGDEASGCHYGALTCGSCKVFFKRAAEGKQKYLCASRNDCTIDKFRRKNCPSCRLRKCYEAGMTLGARKLKKLGNLKLQEEGEASSTTSPTEETTQKLTVSHIEGYECQPIFLNVLEAIEPGVVCAGHDNNQPDSFAALLSSLNELGERQLVHVVKWAKALPGFRNLHVDDQMAVIQYSWMGLMVFAMGWRSFTNVNSRMLYFAPDLVFNEYRMHKSRMYSQCVRMRHLSQEFGWLQITPQEFLCMKALLLFSIIPVDGLKNQKFFDELRMNYIKELDRIIACKRKNPTSCSRRFYQLTKLLDSVQPIARELHQFTFDLLIKSHMVSVDFPEMMAEIISVQVPKILSGKVKPIYFHTQ gak_human Cyclin-G-associated kinase GAK protein protein human Homo sapiens Human MSLLQSALDFLAGPGSLGGASGRDQSDFVGQTVELGELRLRVRRVLAEGGFAFVYEAQDVGSGREYALKRLLSNEEEKNRAIIQEVCFMKKLSGHPNIVQFCSAASIGKEESDTGQAEFLLLTELCKGQLVEFLKKMESRGPLSCDTVLKIFYQTCRAVQHMHRQKPPIIHRDLKVENLLLSNQGTIKLCDFGSATTISHYPDYSWSAQRRALVEEEITRNTTPMYRTPEIIDLYSNFPIGEKQDIWALGCILYLLCFRQHPFEDGAKLRIVNGKYSIPPHDTQYTVFHSLIRAMLQVNPEERLSIAEVVHQLQEIAAARNVNPKSPITELLEQNGGYGSATLSRGPPPPVGPAGSGYSGGLALAEYDQPYGGFLDILRGGTERLFTNLKDTSSKVIQSVANYAKGDLDISYITSRIAVMSFPAEGVESALKNNIEDVRLFLDSKHPGHYAVYNLSPRTYRPSRFHNRVSECGWAARRAPHLHTLYNICRNMHAWLRQDHKNVCVVHCMDGRAASAVAVCSFLCFCRLFSTAEAAVYMFSMKRCPPGIWPSHKRYIEYMCDMVAEEPITPHSKPILVRAVVMTPVPLFSKQRSGCRPFCEVYVGDERVASTSQEYDKMRDFKIEDGKAVIPLGVTVQGDVLIVIYHARSTLGGRLQAKMASMKMFQIQFHTGFVPRNATTVKFAKYDLDACDIQEKYPDLFQVNLEVEVEPRDRPSREAPPWENSSMRGLNPKILFSSREEQQDILSKFGKPELPRQPGSTAQYDAGAGSPEAEPTDSDSPPSSSADASRFLHTLDWQEEKEAETGAENASSKESESALMEDRDESEVSDEGGSPISSEGQEPRADPEPPGLAAGLVQQDLVFEVETPAVLPEPVPQEDGVDLLGLHSEVGAGPAVPPQACKAPSSNTDLLSCLLGPPEAASQGPPEDLLSEDPLLLASPAPPLSVQSTPRGGPPAAADPFGPLLPSSGNNSQPCSNPDLFGEFLNSDSVTVPPSFPSAHSAPPPSCSADFLHLGDLPGEPSKMTASSSNPDLLGGWAAWTETAASAVAPTPATEGPLFSPGGQPAPCGSQASWTKSQNPDPFADLGDLSSGLQGSPAGFPPGGFIPKTATTPKGSSSWQTSRPPAQGASWPPQAKPPPKACTQPRPNYASNFSVIGAREERGVRAPSFAQKPKVSENDFEDLLSNQGFSSRSDKKGPKTIAEMRKQDLAKDTDPLKLKLLDWIEGKERNIRALLSTLHTVLWDGESRWTPVGMADLVAPEQVKKHYRRAVLAVHPDKAAGQPYEQHAKMIFMELNDAWSEFENQGSRPLF andr_rat Androgen receptor Ar Nr3c4 Dihydrotestosterone receptor Nuclear receptor subfamily 3 group C member 4 protein protein rat Rattus norvegicus (Rat) Rat MEVQLGLGRVYPRPPSKTYRGAFQNLFQSVREAIQNPGPRHPEAASIAPPGACLQQRQETSPRRRRRQQHPEDGSPQAHIRGTTGYLALEEEQQPSQQQSASEGHPESGCLPEPGAATAPGKGLPQQPPAPPDQDDSAAPSTLSLLGPTFPGLSSCSADIKDILSEAGTMQLLQQQQQQQQQQQQQQQQQQQQQQEVISEGSSSVRAREATGAPSSSKDSYLGGNSTISDSAKELCKAVSVSMGLGVEALEHLSPGEQLRGDCMYASLLGGPPAVRPTPCAPLAECKGLSLDEGPGKGTEETAEYSSFKGGYAKGLEGESLGCSGSSEAGSSGTLEIPSSLSLYKSGAVDEAAAYQNRDYYNFPLALSGPPHPPPPTHPHARIKLENPSDYGSAWAAAAAQCRYGDLASLHGGSVAGPSTGSPPATASSSWHTLFTAEEGQLYGPGGGGGSSSPSDAGPVAPYGYTRPPQGLASQEGDFSASEVWYPGGVVNRVPYPSPSCVKSEMGPWMENYSGPYGDMRLDSTRDHVLPIDYYFPPQKTCLICGDEASGCHYGALTCGSCKVFFKRAAEGKQKYLCASRNDCTIDKFRRKNCPSCRLRKCYEAGMTLGARKLKKLGNLKLQEEGENSSAGSPTEDPSQKMTVSHIEGYECQPIFLNVLEAIEPGVVCAGHDNNQPDSFAALLSSLNELGERQLVHVVKWAKALPGFRNLHVDDQMAVIQYSWMGLMVFAMGWRSFTNVNSRMLYFAPDLVFNEYRMHKSRMYSQCVRMRHLSQEFGWLQITPQEFLCMKALLLFSIIPVDGLKNQKFFDELRMNYIKELDRIIACKRKNPTSCSRRFYQLTKLLDSVQPIARELHQFTFDLLIKSHMVSVDFPEMMAEIISVQVPKILSGKVKPIYFHTQ ar-ddc DDC and AR interact in a yeast two-hybrid assay 722544 N/A 722547 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 722548 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false PREY gene NM_000790 ar-gak Cyclin G-associated kinase and AR interact in a yeast two-hybrid assay 722544 N/A 722549 unspecified role unspecified role prey prey N/A 722548 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false ar-ddc-2 GST-AR pulls down DDC in vitro 722545 N/A 722547 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false in vitro In vitro N/A 722550 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false 3 The interaction strength for DDC with the AR constructs varied in strength: AR (541-919)> AR(1-559) > AR (234-665) > AR (541-665). This indicated the strongest interaction occured between DDC and the AR LBD. ar-ddc-3 Endogenous AR coimmunoprecipitates endogenous DDC from LNCaP lysates 722546 N/A 722547 unspecified role unspecified role prey prey N/A 722548 unspecified role unspecified role bait bait physical association physical association false false false 4 B Androgen (R1881) treatment enhanced this interacton.