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/ GRIM-19 associates with the serine protease HtrA2 for promoting cell death. GRIM-19 associates with the serine protease HtrA2 for promoting cell death. Oncogene (0950-9232) 2007 imex curation Ma X., Kalakonda S., Srinivasula SM., Reddy SP., Platanias LC., Kalvakolanu DV. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer dkalvako@umaryland.edu 07-Sept-2007: Contacted by JYOTI. Only protein-protein interactions human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Ma X., Kalakonda S., Srinivasula SM., Reddy SP., Platanias LC., Kalvakolanu DV. Parkinsons - Interactions investigated in the context of Parkinsons disease dkalvako@umaryland.edu Accepted 2007-SEP-2 by JYOTI. 07-Sept-2007: Contacted by JYOTI. Oncogene (0950-9232) 2007 Only protein-protein interactions imex curation GRIM-19 associates with the serine protease HtrA2 for promoting cell death. GRIM-19 associates with the serine protease HtrA2 for promoting cell death. Oncogene (0950-9232) 2007 imex curation Ma X., Kalakonda S., Srinivasula SM., Reddy SP., Platanias LC., Kalvakolanu DV. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer dkalvako@umaryland.edu 07-Sept-2007: Contacted by JYOTI. 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 Ma X., Kalakonda S., Srinivasula SM., Reddy SP., Platanias LC., Kalvakolanu DV. Parkinsons - Interactions investigated in the context of Parkinsons disease dkalvako@umaryland.edu Accepted 2007-SEP-2 by JYOTI. 07-Sept-2007: Contacted by JYOTI. Oncogene (0950-9232) 2007 Only protein-protein interactions imex curation GRIM-19 associates with the serine protease HtrA2 for promoting cell death. GRIM-19 associates with the serine protease HtrA2 for promoting cell death. Oncogene (0950-9232) 2007 imex curation Ma X., Kalakonda S., Srinivasula SM., Reddy SP., Platanias LC., Kalvakolanu DV. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer dkalvako@umaryland.edu 07-Sept-2007: Contacted by JYOTI. Only protein-protein interactions human-htert_hme1 Homo sapiens non-oncogenic immortalized mammary epithelial cell line hTERT-HME1 htert_hme1 hTERT-HME1 non-oncogenic immortalized mammary epithelial cell line ATCC Number: CRL-4010 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Ma X., Kalakonda S., Srinivasula SM., Reddy SP., Platanias LC., Kalvakolanu DV. Parkinsons - Interactions investigated in the context of Parkinsons disease dkalvako@umaryland.edu Accepted 2007-SEP-2 by JYOTI. 07-Sept-2007: Contacted by JYOTI. Oncogene (0950-9232) 2007 Only protein-protein interactions imex curation GRIM-19 associates with the serine protease HtrA2 for promoting cell death. GRIM-19 associates with the serine protease HtrA2 for promoting cell death. Oncogene (0950-9232) 2007 imex curation Ma X., Kalakonda S., Srinivasula SM., Reddy SP., Platanias LC., Kalvakolanu DV. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer dkalvako@umaryland.edu 07-Sept-2007: Contacted by JYOTI. Only protein-protein interactions human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Ma X., Kalakonda S., Srinivasula SM., Reddy SP., Platanias LC., Kalvakolanu DV. Parkinsons - Interactions investigated in the context of Parkinsons disease dkalvako@umaryland.edu Accepted 2007-SEP-2 by JYOTI. 07-Sept-2007: Contacted by JYOTI. Oncogene (0950-9232) 2007 Only protein-protein interactions imex curation GRIM-19 associates with the serine protease HtrA2 for promoting cell death. GRIM-19 associates with the serine protease HtrA2 for promoting cell death. Oncogene (0950-9232) 2007 imex curation Ma X., Kalakonda S., Srinivasula SM., Reddy SP., Platanias LC., Kalvakolanu DV. Neurodegeneration - Publications depicting interactions involved in neurodegenerative diseases. Parkinsons - Interactions investigated in the context of Parkinsons disease Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer dkalvako@umaryland.edu 07-Sept-2007: Contacted by JYOTI. Only protein-protein interactions human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Ma X., Kalakonda S., Srinivasula SM., Reddy SP., Platanias LC., Kalvakolanu DV. Parkinsons - Interactions investigated in the context of Parkinsons disease dkalvako@umaryland.edu Accepted 2007-SEP-2 by JYOTI. 07-Sept-2007: Contacted by JYOTI. Oncogene (0950-9232) 2007 Only protein-protein interactions imex curation htra2_human Serine protease HTRA2, mitochondrial High temperature requirement protein A2 Omi stress-regulated endoprotease Serine proteinase OMI Serine protease 25 HTRA2 PRSS25 OMI protein protein human Homo sapiens Human MAAPRAGRGAGWSLRAWRALGGIRWGRRPRLTPDLRALLTSGTSDPRARVTYGTPSLWARLSVGVTEPRACLTSGTPGPRAQLTAVTPDTRTREASENSGTRSRAWLAVALGAGGAVLLLLWGGGRGPPAVLAAVPSPPPASPRSQYNFIADVVEKTAPAVVYIEILDRHPFLGREVPISNGSGFVVAADGLIVTNAHVVADRRRVRVRLLSGDTYEAVVTAVDPVADIATLRIQTKEPLPTLPLGRSADVRQGEFVVAMGSPFALQNTITSGIVSSAQRPARDLGLPQTNVEYIQTDAAIDFGNSGGPLVNLDGEVIGVNTMKVTAGISFAIPSDRLREFLHRGEKKNSSSGISGSQRRYIGVMMLTLSPSILAELQLREPSFPDVQHGVLIHKVILGSPAHRAGLRPGDVILAIGEQMVQNAEDVYEAVRTQSQLAVQIRRGRETLTLYVTPEVTE CEA955A7D0DD8C0D nduad_human NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 13 NDUFA13 CGI-39 CDA016 NADH-ubiquinone oxidoreductase B16.6 subunit Complex I-B16.6 Gene associated with retinoic and interferon-induced mortality 19 protein Cell death regulatory protein GRIM-19 GRIM19 protein protein human Homo sapiens Human MAASKVKQDMPPPGGYGPIDYKRNLPRRGLSGYSMLAIGIGTLIYGHWSIMKWNRERRRLQIEDFEARIALLPLLQAETDRRTLQMLRENLEEEAIIMKDVPDWKVGESVFHTTRWVPPLIGELYGLRTTEEALHASHGFMWYT DIP protein Q9P0J0 original sequence version: null DIP protein Q9P0J0 original sequence version: 100 058F608B235FF856 nduf13-htra2 2 7 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey region PDZ domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association S1 ndufa13-htra2 5 7 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 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 region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 1 C we co-transfected expression vectors that can produce the flag(f)-epitopetagged FL and ND133 HtrA2 proteins, and a myc epitope- tagged GRIM-19 protein into MCF-7 cells. Cell lysates were immunoprecipitated (IPed) using mycepitope tag-specific antibodies and then subjected to a Western blot (WB) analysis with f-tag-specific antibodies Q9P0J0; O43464 transfection ndufa13-htra2-1 4 6 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 7 unspecified role unspecified role bait bait physical association physical association 1 E To determine if endogenous proteins also interact, MCF-7 cell extracts were IPed using GRIM-19-specific (Figure 1e) or isotype control antibodies, and the products were subjected to WB analysis with HtrA2-specific antibodies. The GRIM-19 antibody, but not the isotype control, IPed endogenous HtrA2. Conversely, GRIM-19 was coIPed by HtrA2-specific IFN-beta/Retinoic acid treatment significantly increased this interaction, although IFN-beta alone also pormoted the interaction. htra2-ndufa13 4 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association 1 G To determine if endogenous proteins also interact, MCF-7 cell extracts were IPed using GRIM-19-specific (Figure 1e) or isotype control antibodies, and the products were subjected to WB analysis with HtrA2-specific antibodies. The GRIM-19 antibody, but not the isotype control, IPed endogenous HtrA2. Conversely, GRIM-19 was coIPed by HtrA2-specific htra2-ndufa13-1 3 6 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey physical association physical association S2 A, 7 A IFN-beta/Retinoic acid treatment significantly enhanced this interaction. GRIM-19:HtrA2 interactions also occur in non-tumorigenic mammary epithelial cells Human herpes virus-8-coded oncoprotein vIRF1 inhibited the IFN-beta/Retinoic acid-induced HtrA2-GRIM-19 interaction. ndufa13-htra2-2 1 7 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey physical association physical association Q9P0J0 transfection 3 K we transfected GRIM-19 into MCF-7 cells and stimulated with IFN/RA for various time points. Cell lysates were IPed with myc-tag specific antibodies and the products were subjected to WB analysis with HtrA2-specific antibodies (Figure 3k and m). IFN/RA stimulated GRIM-19s interaction with endogenous HtrA2 IFN-beta/Retinoic acid stimulated GRIM-19's interaction with endogenous HtrA2.