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/ Receptor for activated C-kinase (RACK-1), a WD motif-containing protein, specifically associates with the human type I IFN receptor. Receptor for activated C-kinase (RACK-1), a WD motif-containing protein, specifically associates with the human type I IFN receptor. J. Immunol. (0022-1767) 2000 imex curation Croze E., Usacheva A., Asarnow D., Minshall RD., Perez HD., Colamonici O. Apoptosis - Interactions involving proteins with a function related to apoptosis Synapse - Interactions of proteins with an established role in the presynapse. ed_croze@berlex.com Human B-cell Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Croze E., Usacheva A., Asarnow D., Minshall RD., Perez HD., Colamonici O. Apoptosis - Interactions involving proteins with a function related to apoptosis ed_croze@berlex.com Accepted 2006-JUN-28 by JYOTI. Human B-cell J. Immunol. (0022-1767) 2000 Only protein-protein interactions imex curation Receptor for activated C-kinase (RACK-1), a WD motif-containing protein, specifically associates with the human type I IFN receptor. Receptor for activated C-kinase (RACK-1), a WD motif-containing protein, specifically associates with the human type I IFN receptor. J. Immunol. (0022-1767) 2000 imex curation Croze E., Usacheva A., Asarnow D., Minshall RD., Perez HD., Colamonici O. Apoptosis - Interactions involving proteins with a function related to apoptosis Synapse - Interactions of proteins with an established role in the presynapse. ed_croze@berlex.com Human B-cell 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 Croze E., Usacheva A., Asarnow D., Minshall RD., Perez HD., Colamonici O. Apoptosis - Interactions involving proteins with a function related to apoptosis Directed two-hybrid. ed_croze@berlex.com Accepted 2006-JUN-28 by JYOTI. Human B-cell J. Immunol. (0022-1767) 2000 Only protein-protein interactions imex curation Receptor for activated C-kinase (RACK-1), a WD motif-containing protein, specifically associates with the human type I IFN receptor. Receptor for activated C-kinase (RACK-1), a WD motif-containing protein, specifically associates with the human type I IFN receptor. J. Immunol. (0022-1767) 2000 imex curation Croze E., Usacheva A., Asarnow D., Minshall RD., Perez HD., Colamonici O. Apoptosis - Interactions involving proteins with a function related to apoptosis Synapse - Interactions of proteins with an established role in the presynapse. ed_croze@berlex.com Human B-cell Only protein-protein interactions human-daudi Homo sapiens Burkitt's B-cell lymphona cells daudi Homo sapies DAUDI Burkitt's B-cell lymphoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Croze E., Usacheva A., Asarnow D., Minshall RD., Perez HD., Colamonici O. Apoptosis - Interactions involving proteins with a function related to apoptosis ed_croze@berlex.com Accepted 2006-JUN-28 by JYOTI. Human B-cell J. Immunol. (0022-1767) 2000 Only protein-protein interactions imex curation Receptor for activated C-kinase (RACK-1), a WD motif-containing protein, specifically associates with the human type I IFN receptor. Receptor for activated C-kinase (RACK-1), a WD motif-containing protein, specifically associates with the human type I IFN receptor. J. Immunol. (0022-1767) 2000 imex curation Croze E., Usacheva A., Asarnow D., Minshall RD., Perez HD., Colamonici O. Apoptosis - Interactions involving proteins with a function related to apoptosis Synapse - Interactions of proteins with an established role in the presynapse. ed_croze@berlex.com Human B-cell 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 Croze E., Usacheva A., Asarnow D., Minshall RD., Perez HD., Colamonici O. Apoptosis - Interactions involving proteins with a function related to apoptosis ed_croze@berlex.com Accepted 2006-JUN-28 by JYOTI. Human B-cell J. Immunol. (0022-1767) 2000 Only protein-protein interactions imex curation inar2_human Interferon alpha/beta receptor 2 IFNAR2 IFNARB Type I interferon receptor 2 IFNABR Interferon alpha binding protein protein protein human Homo sapiens Human MLLSQNAFIFRSLNLVLMVYISLVFGISYDSPDYTDESCTFKISLRNFRSILSWELKNHSIVPTHYTLLYTIMSKPEDLKVVKNCANTTRSFCDLTDEWRSTHEAYVTVLEGFSGNTTLFSCSHNFWLAIDMSFEPPEFEIVGFTNHINVMVKFPSIVEEELQFDLSLVIEEQSEGIVKKHKPEIKGNMSGNFTYIIDKLIPNTNYCVSVYLEHSDEQAVIKSPLKCTLLPPGQESESAESAKIGGIITVFLIALVLTSTIVTLKWIGYICLRNSLPKVLNFHNFLAWPFPNLPPLEAMDMVEVIYINRKKKVWDYNYDDESDSDTEAAPRTSGGGYTMHGLTVRPLGQASATSTESQLIDPESEEEPDLPEVDVELPTMPKDSPQQLELLSGPCERRKSPLQDPFPEEDYSSTEGSGGRITFNVDLNSVFLRVLDDEDSDDLEAPLMLSSHLEEMVDPEDPDNVQSNHLLASGEGTQPTFPSPSSEGLWSEDAPSDQSDTSESDVDLGDGYIMR 4D7730D93AA739F4 rack1_human Receptor of activated protein C kinase 1 Small ribosomal subunit protein RACK1 HLC7 PIG21 RACK1 Cell proliferation-inducing gene 21 protein GNB2L1 Guanine nucleotide-binding protein subunit beta-2-like 1 Guanine nucleotide-binding protein subunit beta-like protein 12.3 Human lung cancer oncogene 7 protein Receptor for activated C kinase protein protein human Homo sapiens Human MTEQMTLRGTLKGHNGWVTQIATTPQFPDMILSASRDKTIIMWKLTRDETNYGIPQRALRGHSHFVSDVVISSDGQFALSGSWDGTLRLWDLTTGTTTRRFVGHTKDVLSVAFSSDNRQIVSGSRDKTIKLWNTLGVCKYTVQDESHSEWVSCVRFSPNSSNPIIVSCGWDKLVKVWNLANCKLKTNHIGHTGYLNTVTVSPDGSLCASGGKDGQAMLWDLNEGKHLYTLDGGDIINALCFSPNRYWLCAATGPSIKIWDLEGKIIVDELKQEVISTSSKAEPPQCTSLAWSADGQTLFAGYTDNLVRVWQVTIGTR DIP protein P63244 original sequence version: 71 257F91E369ED2044 ifnar2-gnb2l1-1 4 6 unspecified role unspecified role prey prey c-terminal region sufficient to bind sufficient binding region sufficient to bind full dna sequence full identification by DNA sequencing full dna sequence certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role bait bait cytoplasmic region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 9 11 physical association physical association Yeast two-hybrid interaction of RACK-1 with the cytoplasmic domain the IFN-R�L chain. 1 ifnar2-gnb2l1-2 1 6 unspecified role unspecified role prey prey human-lymphoblast Homo sapiens, lymphoblast lymphoblast Transformed lymphocytes; Blast cell; Lymphoblastoid 5 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 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) physical association physical association To confirm the interaction between IFN-R�L and RACK-1, we performed "pull-down" experiments. GST constructs containing the entire cytoplasmic domain of IFN-R�L bound RACK-1 in IFN-stimulated or unstimulated cell lysates (Fig. 2A). 2 ifnar2-gnb2l1-3 3 5 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association Coimmunoprecipitation of RACK-1 with IFN-R�L is independent of IFN stimulation. 3 ifnar2-gnb2l1-4 2 6 unspecified role unspecified role prey prey c-terminal region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role bait bait cytoplasmic region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association The interaction between RACK-1 and IFN-R�L detected in the two-hybrid system was shown to be specific, because both the pACT-C1-L and pACT-C2-N1 clones failed to produce �-galactosidase-positive colonies when either was reintroduced with one of two different control plasmids (pLAM 5'-1 or pVA3-1) in CG1945 cells (Fig. 1). Cotransformation of the control (pLAM 5'-1 or pVA3-1) and bait plasmids (pACT-C1-L or pACT-C2-N1) was confirmed by the observed growth of transformed yeast in tryptophan- and leucine-deficient media (Fig. 1). 1B