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/ Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Exp. Cell Res. (0014-4827) 2005 Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer The source of Par-4 was taken from PMID 8043520. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. orchard rat-ref52 Rattus norvegicus embryonic fibroblast cells ref52 Rattus norvegicus embryonic fibroblast cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis The interactions in the heterokaryons were not curated as it is not clear to me that it is possible to add this data. The source of Par-4 was taken from PMID 8043520. The source of Par-4 was taken from PMID 8043520. Accepted 2008-MAY-30 by ORCHARD. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. Exp. Cell Res. (0014-4827) 2005 orchard Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Exp. Cell Res. (0014-4827) 2005 Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer The source of Par-4 was taken from PMID 8043520. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. orchard 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 Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of Par-4 was taken from PMID 8043520. Accepted 2008-MAY-30 by ORCHARD. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. Exp. Cell Res. (0014-4827) 2005 orchard Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Exp. Cell Res. (0014-4827) 2005 Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer The source of Par-4 was taken from PMID 8043520. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. orchard rat-ref52 Rattus norvegicus embryonic fibroblast cells ref52 Rattus norvegicus embryonic fibroblast cell line fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of Par-4 was taken from PMID 8043520. Accepted 2008-MAY-30 by ORCHARD. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. Exp. Cell Res. (0014-4827) 2005 orchard Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Exp. Cell Res. (0014-4827) 2005 Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer The source of Par-4 was taken from PMID 8043520. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. orchard crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of Par-4 was taken from PMID 8043520. Accepted 2008-MAY-30 by ORCHARD. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. Exp. Cell Res. (0014-4827) 2005 orchard Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Exp. Cell Res. (0014-4827) 2005 Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer The source of Par-4 was taken from PMID 8043520. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. orchard crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of Par-4 was taken from PMID 8043520. Accepted 2008-MAY-30 by ORCHARD. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. Exp. Cell Res. (0014-4827) 2005 orchard Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Exp. Cell Res. (0014-4827) 2005 Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer The source of Par-4 was taken from PMID 8043520. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. orchard in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of Par-4 was taken from PMID 8043520. Accepted 2008-MAY-30 by ORCHARD. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. Exp. Cell Res. (0014-4827) 2005 orchard Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Par-4-mediated recruitment of Amida to the actin cytoskeleton leads to the induction of apoptosis. Exp. Cell Res. (0014-4827) 2005 Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer The source of Par-4 was taken from PMID 8043520. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. orchard 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 Boosen M., Vetterkind S., Koplin A., Illenberger S., Preuss U. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of Par-4 was taken from PMID 8043520. Accepted 2008-MAY-30 by ORCHARD. A cDNA library generated from SV52 cells was used. SV52 cells are SV40 large T-transformed rat embryo fibroblasts. Exp. Cell Res. (0014-4827) 2005 orchard pawr_rat PRKC apoptosis WT1 regulator protein Prostate apoptosis response 4 protein Transcriptional repressor Par-4-like protein PAWR Pawr Par4 protein protein rat Rattus norvegicus (Rat) Rat MATGGYRSSGSTTDFLEEWKAKREKMRAKQNPVGPGSSGGDPAAKSPAGPLAQTTAAGTSELNHGPAGAAAPAAPGPGALNCAHGSSALPRGAPGSRRPEDECPIAAGAAGAPASRGDEEEPDSAPEKGRSSGPSARKGKGQIEKRKLREKRRSTGVVNIPAAECLDEYEDDEAGQKERKREDAITQQNTIQNEAASLPDPGTSYLPQDPSRTVPGRYKSTISAPEEEILNRYPRTDRSGFSRHNRDTSAPANFASSSTLEKRIEDLEKEVLRERQENLRLTRLMQDKEEMIGKLKEEIDLLNRDLDDMEDENEQLKQENKTLLKVVGQLTR 2069B32DEFFF160F tfpt_rat TCF3 fusion partner homolog Tfpt Amida Protein amida protein protein rat Rattus norvegicus (Rat) Rat MELEQREGTMAAVGFEEFSAPPGSELALPPLFGGHILESELETEVEFVSGGLGDSGLRERDEEEEAARGRRRRQRELNRRKYQALGRRCREIEQVNERVLNRLHQVQRITRRLQQERRFLMRVLDSYGDDYRDSQFTIVLEDDGSQGTDVPTPGNVENEPPEKEGLSPPQRTTATLDPSSPAPGEGPSGRKRRRAPRAASSLTPELAPVQVGAEGWGQGVIKVEEDFGFEADEALDSSWVSRGPDKLLPYPTLASPPFD 674408B3F3FB6863 pawr-pawr 7 8 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association we used Par-4 as bait to screen a cDNA library from SV52 rat fibroblasts. Seven positive clones were identified from histidine negative plates which were also positive for h-galactosidase expression. Sequence analysis revealed one clone coding for Par-4, showing the ability of the protein to self-associate, pawr-tfpt 7 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association Using Par-4 as bait in a yeast two-hybrid screen, we identified Amida as a novel interaction partner pawr-tfpt-1 2 9 unspecified role unspecified role prey prey c-terminal required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role bait bait physical association physical association The interaction between Par-4 and Amida was first confirmed by retransformation of yeast cells with plasmids coding for pAD-Amida and pBD-Par-4 including the appropriate controls 1 B pawr-tfpt-2 6 9 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 in vitro In vitro 8 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction For this purpose, Par-4 was expressed as a GST-tagged fusion protein in E. coli, immobilized on glutathione sepharose beads and used as affinity matrix to bind 35S-methionine-labeled in-vitro-translated Amida. 2 A tfpt-pawr 4 9 unspecified role unspecified role bait bait region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey 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 2 B Q62627; Q9JMG6 transfection Immunoprecipitation of GFP-Amida with an anti-GFP antibody specifically coprecipitated a protein of about 48 kDa representing the FLAG-Par-4 fusion protein pawr-tfpt-3 5 9 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role bait bait 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 2 B Q62627; Q9JMG6 transfection when employing anti-FLAG specific antibodies, a protein of about 68 kDa corresponding to the GFP-Amida fusion protein (Fig. 2B, lane 4) was only present in the precipitate from FLAGPar-4-expressing cells but not from mock-transfected cells tfpt-pawr-1 1 9 unspecified role unspecified role bait bait region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey physical association physical association 2 C Q9JMG6 transfection By employing the anti-Amida antibody, we could coprecipitate endogenous Par-4 protein tfpt-pawr-2 3 8 unspecified role unspecified role prey prey 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 9 unspecified role unspecified role bait bait region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization Q9JMG6; Q62627 transfection 4 GFPAmida was no longer diffusely distributed in the cytoplasm but clearly colocalized with Par-4 at actin filaments in the coexpressing cells This interaction took place at actin filaments. pawr-tfpt-4 2 8 unspecified role unspecified role bait bait Q62627:p.[Leu295Ala;Leu316Ala;Leu330Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L A certain certain sequence position certain certain certain sequence position certain L A certain certain sequence position certain certain certain sequence position certain L A Wed May 02 19:29:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu295ala,leu316ala,leu330ala' Tue Jun 30 17:36:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu295Ala;Leu316Ala;Leu330Ala]' c-terminal required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 9 unspecified role unspecified role prey prey physical association physical association The interaction between Par-4 and Amida was first confirmed by retransformation of yeast cells with plasmids coding for pAD-Amida and pBD-Par-4 including the appropriate controls 1 B