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/ chiocchetti-2005-3 Interactions between RPS19, mutated in Diamond-Blackfan anemia, and the PIM-1 oncoprotein. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Chiocchetti A., Gibello L., Carando A., Aspesi A., Secco P., Garelli E., Loreni F., Angelini M., Biava A., Dahl N., Dianzani U., Ramenghi U., Santoro C., Dianzani I. irma.dianzani@ med.unipmn.it Haematologica (0390-6078) 2005 Only protein-protein interactions imex curation chiocchetti-2005-1 Interactions between RPS19, mutated in Diamond-Blackfan anemia, and the PIM-1 oncoprotein. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Chiocchetti A., Gibello L., Carando A., Aspesi A., Secco P., Garelli E., Loreni F., Angelini M., Biava A., Dahl N., Dianzani U., Ramenghi U., Santoro C., Dianzani I. irma.dianzani@ med.unipmn.it Haematologica (0390-6078) 2005 Only protein-protein interactions imex curation chiocchetti-2005-2 Interactions between RPS19, mutated in Diamond-Blackfan anemia, and the PIM-1 oncoprotein. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Chiocchetti A., Gibello L., Carando A., Aspesi A., Secco P., Garelli E., Loreni F., Angelini M., Biava A., Dahl N., Dianzani U., Ramenghi U., Santoro C., Dianzani I. irma.dianzani@ med.unipmn.it Haematologica (0390-6078) 2005 Only protein-protein interactions imex curation chiocchetti-2005-4 Interactions between RPS19, mutated in Diamond-Blackfan anemia, and the PIM-1 oncoprotein. in vitro In vitro in gel kinase assay in-gel kinase assay in gel kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Chiocchetti A., Gibello L., Carando A., Aspesi A., Secco P., Garelli E., Loreni F., Angelini M., Biava A., Dahl N., Dianzani U., Ramenghi U., Santoro C., Dianzani I. irma.dianzani@ med.unipmn.it Haematologica (0390-6078) 2005 Only protein-protein interactions imex curation chiocchetti-2005-5 Interactions between RPS19, mutated in Diamond-Blackfan anemia, and the PIM-1 oncoprotein. yeast Saccharomyces cerevisiae 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 Chiocchetti A., Gibello L., Carando A., Aspesi A., Secco P., Garelli E., Loreni F., Angelini M., Biava A., Dahl N., Dianzani U., Ramenghi U., Santoro C., Dianzani I. irma.dianzani@ med.unipmn.it Haematologica (0390-6078) 2005 Only protein-protein interactions imex curation chiocchetti-2005-6 Interactions between RPS19, mutated in Diamond-Blackfan anemia, and the PIM-1 oncoprotein. yeast Saccharomyces cerevisiae 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 Chiocchetti A., Gibello L., Carando A., Aspesi A., Secco P., Garelli E., Loreni F., Angelini M., Biava A., Dahl N., Dianzani U., Ramenghi U., Santoro C., Dianzani I. cDNA library of human embryonic liver cells obtained at 9-24 weeks of gestation, when hepatic erythropoiesis is active. irma.dianzani@ med.unipmn.it Haematologica (0390-6078) 2005 Only protein-protein interactions imex curation rs19_human 40S ribosomal protein S19 RPS19 Small ribosomal subunit protein eS19 protein protein human Homo sapiens Human MPGVTVKDVNQQEFVRALAAFLKKSGKLKVPEWVDTVKLAKHKELAPYDENWFYTRAASTARHLYLRGGAGVGSMTKIYGGRQRNGVMPSHFSRGSKSVARRVLQALEGLKMVEKDQDGGRKLTPQGQRDLDRIAGQVAAANKKH pim1_human Serine/threonine-protein kinase pim-1 PIM1 protein protein human Homo sapiens Human MLLSKINSLAHLRAAPCNDLHATKLAPGKEKEPLESQYQVGPLLGSGGFGSVYSGIRVSDNLPVAIKHVEKDRISDWGELPNGTRVPMEVVLLKKVSSGFSGVIRLLDWFERPDSFVLILERPEPVQDLFDFITERGALQEELARSFFWQVLEAVRHCHNCGVLHRDIKDENILIDLNRGELKLIDFGSGALLKDTVYTDFDGTRVYSPPEWIRYHRYHGRSAAVWSLGILLYDMVCGDIPFEHDEEIIRGQVFFRQRVSSECQHLIRWCLALRPSDRPTFEEIQNHPWMQDVLLPQETAEIHLHSLSPGPSK rps19-pim1-1 1365648 n/a 1365654 unspecified role unspecified role bait bait P39019:p.Arg62Trp mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false P39019:p.Arg101His mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false gst 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 ecoli Escherichia coli (strain K12) n/a 1365655 unspecified role unspecified role prey prey human-k_562 Homo sapiens myeloid leukemia cells. k562 Myelogenous leukemia K562 cells The pull-down assay was performed with K562 whole cell extracts physical association physical association false false false Figure 1C, Figure 3C pim1-rps19-1 1365648 n/a 1365655 unspecified role unspecified role bait bait gst 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 ecoli Escherichia coli (strain K12) n/a 1365654 unspecified role unspecified role prey prey human Homo sapiens Human physical association physical association false false false Fig 1C pim1-rps19-5 1365649 n/a 1365655 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation gst gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag full dna sequence full identification by DNA sequencing full dna sequence undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1365654 unspecified role unspecified role prey prey nucl transfection nucleic acid transfection nucl transfection flag-tag 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 false P39019:p.Arg101His mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P39019:p.Arg56Gln mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false P39019:p.Arg62Trp mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. physical association physical association false false false Figure 1A, Figure 3A RPS19 protein was produced in human embryonic kidney 293T cells transfected with pFLAGRPS19 plasmid DNA rps19-pim1 1365650 n/a 1365655 unspecified role unspecified role prey prey nucl transfection nucleic acid transfection nucl transfection ha ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1365654 unspecified role unspecified role bait bait nucl transfection nucleic acid transfection nucl transfection flag tag 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 false physical association physical association false false false Figure 1B 293T cells were co-transfected with pFLAG-RPS19 and pHA-PIM DNA pim1-rps19-4 1365651 n/a 1365655 enzyme enzyme neutral component neutral component nucl transformation nucleic acid transformation nucl transformation gst 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 ecoli Escherichia coli (strain K12) n/a 1365654 enzyme target enzyme target substrate substrate neutral component neutral component nucl transformation nucleic acid transformation nucl transformation gst 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 ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false Fig 2 rps19-pim1-2 1365652 n/a 1365654 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation P39019:p.Arg101His mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P39019:p.Arg56Gln mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false P39019:p.Arg62Trp mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false n/a 1365655 unspecified role unspecified role prey prey physical association physical association false false false Figure 3B rps19-pim1-3 1365653 n/a 1365655 unspecified role unspecified role prey prey n/a 1365654 unspecified role unspecified role bait bait physical association physical association false false false