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/ The gene product Murr1 restricts HIV-1 replication in resting CD4+ lymphocytes. The gene product Murr1 restricts HIV-1 replication in resting CD4+ lymphocytes. Nature (0028-0836) 2003 imex curation Ganesh L., Burstein E., Guha-Niyogi A., Louder MK., Mascola JR., Klomp LW., Wijmenga C., Duckett CS., Nabel GJ. The origins of transfected Murr1 determined based on the publications from the author's lab. gnabel@nih.gov 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions 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. pull down pull down anti tag western anti tag western blot anti tag western Ganesh L., Burstein E., Guha-Niyogi A., Louder MK., Mascola JR., Klomp LW., Wijmenga C., Duckett CS., Nabel GJ. gnabel@nih.gov Accepted 2007-DEC-7 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. Nature (0028-0836) 2003 orchard Only protein-protein interactions imex curation The gene product Murr1 restricts HIV-1 replication in resting CD4+ lymphocytes. The gene product Murr1 restricts HIV-1 replication in resting CD4+ lymphocytes. Nature (0028-0836) 2003 imex curation Ganesh L., Burstein E., Guha-Niyogi A., Louder MK., Mascola JR., Klomp LW., Wijmenga C., Duckett CS., Nabel GJ. The origins of transfected Murr1 determined based on the publications from the author's lab. gnabel@nih.gov 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions 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 tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Ganesh L., Burstein E., Guha-Niyogi A., Louder MK., Mascola JR., Klomp LW., Wijmenga C., Duckett CS., Nabel GJ. The origins of transfected Murr1 determined based on the publications from the author's lab. gnabel@nih.gov Accepted 2007-DEC-7 by ORCHARD. Nature (0028-0836) 2003 orchard Only protein-protein interactions imex curation The gene product Murr1 restricts HIV-1 replication in resting CD4+ lymphocytes. The gene product Murr1 restricts HIV-1 replication in resting CD4+ lymphocytes. Nature (0028-0836) 2003 imex curation Ganesh L., Burstein E., Guha-Niyogi A., Louder MK., Mascola JR., Klomp LW., Wijmenga C., Duckett CS., Nabel GJ. The origins of transfected Murr1 determined based on the publications from the author's lab. gnabel@nih.gov 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions 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. confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Ganesh L., Burstein E., Guha-Niyogi A., Louder MK., Mascola JR., Klomp LW., Wijmenga C., Duckett CS., Nabel GJ. gnabel@nih.gov Accepted 2007-DEC-7 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. Nature (0028-0836) 2003 orchard Only protein-protein interactions imex curation The gene product Murr1 restricts HIV-1 replication in resting CD4+ lymphocytes. The gene product Murr1 restricts HIV-1 replication in resting CD4+ lymphocytes. Nature (0028-0836) 2003 imex curation Ganesh L., Burstein E., Guha-Niyogi A., Louder MK., Mascola JR., Klomp LW., Wijmenga C., Duckett CS., Nabel GJ. The origins of transfected Murr1 determined based on the publications from the author's lab. gnabel@nih.gov 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions 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 western blot western blot Immuno blot Immuno blot Ganesh L., Burstein E., Guha-Niyogi A., Louder MK., Mascola JR., Klomp LW., Wijmenga C., Duckett CS., Nabel GJ. gnabel@nih.gov Accepted 2007-DEC-7 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. Nature (0028-0836) 2003 orchard Only protein-protein interactions imex curation tf65_human Transcription factor p65 Nuclear factor NF-kappa-B p65 subunit RELA NFKB3 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 protein protein human Homo sapiens Human MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGSIPGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAELCPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQVTVRDPSGRPLRLPPVLSHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQKEDIEVYFTGPGWEARGSFSQADVHRQVAIVFRTPPYADPSLQAPVRVSMQLRRPSDRELSEPMEFQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISS DIP protein Q04206 original sequence version: 170 DIP protein Q04206 original sequence version: 173 DIP protein Q04206 original sequence version: 99 DIP protein Q04206-1 original sequence version: 99 DIP protein Q04206 original sequence version: 168 DIP protein Q04206 original sequence version: 128 DIP protein Q04206 original sequence version: 131 DIP protein Q04206 original sequence version: 153 DIP protein Q04206 original sequence version: 185 DIP protein Q04206 original sequence version: 181 DIP protein Q04206 original sequence version: 183 8147A6AF9F2445C6 ikbb_human NF-kappa-B inhibitor beta NFKBIB IKBB TRIP9 I-kappa-B-beta Thyroid receptor-interacting protein 9 protein protein human Homo sapiens Human MAGVACLGKAADADEWCDSGLGSLGPDAAAPGGPGLGAELGPGLSWAPLVFGYVTEDGDTALHLAVIHQHEPFLDFLLGFSAGTEYMDLQNDLGQTALHLAAILGETSTVEKLYAAGAGLCVAERRGHTALHLACRVGAHACARALLQPRPRRPREAPDTYLAQGPDRTPDTNHTPVALYPDSDLEKEEEESEEDWKLQLEAENYEGHTPLHVAVIHKDVEMVRLLRDAGADLDKPEPTCGRSPLHLAVEAQAADVLELLLRAGANPAARMYGGRTPLGSAMLRPNPILARLLRAHGAPEPEGEDEKSGPCSSSSDSDSGDEGDEYDDIVVHSSRSQTRLPPTPASKPLPDDPRPV E84575971B6F81CC comd1_human COMM domain-containing protein 1 COMMD1 C2orf5 MURR1 Protein Murr1 protein protein human Homo sapiens Human MAAGELEGGKPLSGLLNALAQDTFHGYPGITEELLRSQLYPEVPPEEFRPFLAKMRGILKSIASADMDFNQLEAFLTAQTKKQGGITSDQAAVISKFWKSHKTKIRESLMNQSRWNSGLRGLSWRVDGKSQSRHSAQIHTPVAIIELELGKYGQESEFLCLEFDEVKVNQILKTLSEVEESISTLISQPN 9C810AECA67011DC skp1_human S-phase kinase-associated protein 1 SKP1 EMC19 OCP2 SKP1A TCEB1L Cyclin-A/CDK2-associated protein p19 p19skp1 RNA polymerase II elongation factor-like protein Organ of Corti protein 2 Organ of Corti protein II SIII Transcription elongation factor B polypeptide 1-like protein protein human Homo sapiens Human MPSIKLQSSDGEIFEVDVEIAKQSVTIKTMLEDLGMDDEGDDDPVPLPNVNAAILKKVIQWCTHHKDDPPPPEDDENKEKRTDDIPVWDQEFLKVDQGTLFELILAANYLDIKGLLDVTCKTVANMIKGKTPEEIRKTFNIKNDFTEEEEAQVRKENQWCEEK C794D62AFB75528A ikba_human NF-kappa-B inhibitor alpha NFKBIA IKBA MAD3 NFKBI I-kappa-B-alpha Major histocompatibility complex enhancer-binding protein MAD3 protein protein human Homo sapiens Human MFQAAERPQEWAMEGPRDGLKKERLLDDRHDSGLDSMKDEEYEQMVKELQEIRLEPQEVPRGSEPWKQQLTEDGDSFLHLAIIHEEKALTMEVIRQVKGDLAFLNFQNNLQQTPLHLAVITNQPEIAEALLGAGCDPELRDFRGNTPLHLACEQGCLASVGVLTQSCTTPHLHSILKATNYNGHTCLHLASIHGYLGIVELLVSLGADVNAQEPCNGRTALHLAVDLQNPDLVSLLLKCGADVNRVTYQGYSPYQLTWGRPSTRIQQQLGQLTLENLQMLPESEDEESYDTESEFTEFTEDELPYDDCVFGGQRLTL DIP protein P25963 original sequence version: 160 DIP protein P25963 original sequence version: 157 DIP protein P25963 original sequence version: 145 DIP protein P25963 original sequence version: 170 DIP protein P25963 original sequence version: 122 DIP protein P25963 original sequence version: 128 DIP protein P25963 original sequence version: 78 DIP protein P25963 original sequence version: 121 DIP protein P25963 original sequence version: 124 088B313226786395 cul1_human Cullin-1 CUL1 protein protein human Homo sapiens Human MSSTRSQNPHGLKQIGLDQIWDDLRAGIQQVYTRQSMAKSRYMELYTHVYNYCTSVHQSNQARGAGVPPSKSKKGQTPGGAQFVGLELYKRLKEFLKNYLTNLLKDGEDLMDESVLKFYTQQWEDYRFSSKVLNGICAYLNRHWVRRECDEGRKGIYEIYSLALVTWRDCLFRPLNKQVTNAVLKLIEKERNGETINTRLISGVVQSYVELGLNEDDAFAKGPTLTVYKESFESQFLADTERFYTRESTEFLQQNPVTEYMKKAEARLLEEQRRVQVYLHESTQDELARKCEQVLIEKHLEIFHTEFQNLLDADKNEDLGRMYNLVSRIQDGLGELKKLLETHIHNQGLAAIEKCGEAALNDPKMYVQTVLDVHKKYNALVMSAFNNDAGFVAALDKACGRFINNNAVTKMAQSSSKSPELLARYCDSLLKKSSKNPEEAELEDTLNQVMVVFKYIEDKDVFQKFYAKMLAKRLVHQNSASDDAEASMISKLKQACGFEYTSKLQRMFQDIGVSKDLNEQFKKHLTNSEPLDLDFSIQVLSSGSWPFQQSCTFALPSELERSYQRFTAFYASRHSGRKLTWLYQLSKGELVTNCFKNRYTLQASTFQMAILLQYNTEDAYTVQQLTDSTQIKMDILAQVLQILLKSKLLVLEDENANVDEVELKPDTLIKLYLGYKNKKLRVNINVPMKTEQKQEQETTHKNIEEDRKLLIQAAIVRIMKMRKVLKHQQLLGEVLTQLSSRFKPRVPVIKKCIDILIEKEYLERVDGEKDTYSYLA DIP protein Q13616 original sequence version: 144 DIP protein Q13616 original sequence version: 133 DIP protein Q13616 original sequence version: null DIP protein Q13616 original sequence version: 135 DIP protein Q13616 original sequence version: 98 DIP protein Q13616 original sequence version: 110 DIP protein Q13616 original sequence version: 146 DIP protein Q13616 original sequence version: 148 DIP protein Q13616 original sequence version: 100 DIP protein Q13616 original sequence version: 85 DIP protein Q13616 original sequence version: 114 DIP protein Q13616 original sequence version: 119 DIP protein Q13616 original sequence version: 136 DIP protein Q13616 original sequence version: 117 DIP protein Q13616 original sequence version: 99 DIP protein Q13616 original sequence version: 108 6625A1FFA7799BBA commd1-ikba 2 9 unspecified role unspecified role prey prey 7 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 ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 2A, Supplementary figure 1 an interaction between transfected HA-tagged Murr1 and endogenous IkappaB-alpha was readily detected The ankyrin domain of IkappaB-alpha was required for association with Murr1, as were amino acids 1-160 of Murr1 rela-ikbb 4 7 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey association association 2B RelA antibody immunoprecipitated IkappaB-alpha, IkappaB-beta and Murr1 ikba-rela 4 9 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey association association 2B IkappaB-alpha antibody also pulled down RelA and Murr1. ikbb-rela 4 6 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey physical association physical association 2B ikba-commd1 1 7 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind range range range range ank repeat required to bind necessary binding region required to bind range range range range physical association physical association Supplementary figure 1 The ankyrin domain of IkappaB-alpha was required for association with Murr1, as were amino acids 1-160 of Murr1 rela-commd1-2 3 7 unspecified role unspecified role neutral component neutral component dsred tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role neutral component neutral component 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 physical association physical association supplementary figure 1B suggesting that Murr1 interacted preferentially with the NF-kappaB�IkappaB-alpha complex. This association was confirmed in vivo by confocal microscopy with fluorescent fusion proteins commd1-cul1 4 7 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association 3C Murr1 interacted biochemically with the Cul1 and not the Skp1 component of the E3 ligase complex cul1-skp1-6 4 8 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association 3C Murr1 interacted biochemically with the Cul1 and not the Skp1 component of the E3 ligase complex