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/ A novel RING finger protein, human enhancer of invasion 10, alters mitotic progression through regulation of cyclin B levels. A novel RING finger protein, human enhancer of invasion 10, alters mitotic progression through regulation of cyclin B levels. Mol. Cell. Biol. (0270-7306) 2003 imex curation Toby GG., Gherraby W., Coleman TR., Golemis EA. Apoptosis - Interactions involving proteins with a function related to apoptosis EA_Golemis@fccc.edu 07-Sept-2007: Contacted by JYOTI. A human fetal brain library was used. davet 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 Toby GG., Gherraby W., Coleman TR., Golemis EA. Apoptosis - Interactions involving proteins with a function related to apoptosis EA_Golemis@fccc.edu Accepted 2007-SEP-2 by DAVET. 07-Sept-2007: Contacted by JYOTI. A human fetal brain library was used. Mol. Cell. Biol. (0270-7306) 2003 davet Only protein-protein interactions imex curation A novel RING finger protein, human enhancer of invasion 10, alters mitotic progression through regulation of cyclin B levels. A novel RING finger protein, human enhancer of invasion 10, alters mitotic progression through regulation of cyclin B levels. Mol. Cell. Biol. (0270-7306) 2003 imex curation Toby GG., Gherraby W., Coleman TR., Golemis EA. Apoptosis - Interactions involving proteins with a function related to apoptosis EA_Golemis@fccc.edu 07-Sept-2007: Contacted by JYOTI. A human fetal brain library was used. davet Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Toby GG., Gherraby W., Coleman TR., Golemis EA. Apoptosis - Interactions involving proteins with a function related to apoptosis EA_Golemis@fccc.edu Accepted 2007-SEP-2 by DAVET. 07-Sept-2007: Contacted by JYOTI. A human fetal brain library was used. Mol. Cell. Biol. (0270-7306) 2003 davet Only protein-protein interactions imex curation A novel RING finger protein, human enhancer of invasion 10, alters mitotic progression through regulation of cyclin B levels. A novel RING finger protein, human enhancer of invasion 10, alters mitotic progression through regulation of cyclin B levels. Mol. Cell. Biol. (0270-7306) 2003 imex curation Toby GG., Gherraby W., Coleman TR., Golemis EA. Apoptosis - Interactions involving proteins with a function related to apoptosis EA_Golemis@fccc.edu 07-Sept-2007: Contacted by JYOTI. A human fetal brain library was used. davet Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Toby GG., Gherraby W., Coleman TR., Golemis EA. Apoptosis - Interactions involving proteins with a function related to apoptosis The authors confirmed by email that all recombinant proteins were of human origin. The materials and methods specify that the recombinant ubiqutin was from Sigma; the Sigma website specifies that human ubiquitin is expressed in E. coli. EA_Golemis@fccc.edu Accepted 2007-SEP-2 by DAVET. 07-Sept-2007: Contacted by JYOTI. A human fetal brain library was used. Mol. Cell. Biol. (0270-7306) 2003 davet Only protein-protein interactions imex curation ccnb1_human G2/mitotic-specific cyclin-B1 CCNB1 CCNB protein protein human Homo sapiens Human MALRVTRNSKINAENKAKINMAGAKRVPTAPAATSKPGLRPRTALGDIGNKVSEQLQAKMPMKKEAKPSATGKVIDKKLPKPLEKVPMLVPVPVSEPVPEPEPEPEPEPVKEEKLSPEPILVDTASPSPMETSGCAPAEEDLCQAFSDVILAVNDVDAEDGADPNLCSEYVKDIYAYLRQLEEEQAVRPKYLLGREVTGNMRAILIDWLVQVQMKFRLLQETMYMTVSIIDRFMQNNCVPKKMLQLVGVTAMFIASKYEEMYPPEIGDFAFVTDNTYTKHQIRQMEMKILRALNFGLGRPLPLHFLRRASKIGEVDVEQHTLAKYLMELTMLDYDMVHFPPSQIAAGAFCLALKILDNGEWTPTLQHYLSYTEESLLPVMQHLAKNVVMVNQGLTKHMTVKNKYATSKHAKISTLPQLNSALVQDLAKAVAKV DIP protein P14635 original sequence version: 146 DIP protein P14635 original sequence version: 108 DIP protein P14635 original sequence version: 145 DIP protein P14635 original sequence version: 139 E2C4767AE8A11EC0 ub2l3_human Ubiquitin-conjugating enzyme E2 L3 Ubiquitin-protein ligase L3 Ubiquitin carrier protein L3 UbcH7 Ubiquitin-conjugating enzyme E2-F1 L-UBC UBE2L3 UBCE7 UBCH7 E2 ubiquitin-conjugating enzyme L3 protein protein human Homo sapiens Human MAASRRLMKELEEIRKCGMKNFRNIQVDEANLLTWQGLIVPDNPPYDKGAFRIEINFPAEYPFKPPKITFKTKIYHPNIDEKGQVCLPVISAENWKPATKTDQVIQSLIALVNDPQPEHPLRADLAEEYSKDRKKFCKNAEEFTKKYGEKRPVD F5A30243BE3C9985 ubiqp_xenla Polyubiquitin protein protein xenla Xenopus laevis (African clawed frog) African clawed frog QKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGG DIP protein P62972 original sequence version: 41 9C8F431544081131 cdk1_human Cyclin-dependent kinase 1 p34 protein kinase Cell division protein kinase 1 CDK1 CDC2 Cell division control protein 2 homolog CDC28A CDKN1 P34CDC2 protein protein human Homo sapiens Human MEDYTKIEKIGEGTYGVVYKGRHKTTGQVVAMKKIRLESEEEGVPSTAIREISLLKELRHPNIVSLQDVLMQDSRLYLIFEFLSMDLKKYLDSIPPGQYMDSSLVKSYLYQILQGIVFCHSRRVLHRDLKPQNLLIDDKGTIKLADFGLARAFGIPIRVYTHEVVTLWYRSPEVLLGSARYSTPVDIWSIGTIFAELATKKPLFHGDSEIDQLFRIFRALGTPNNEVWPEVESLQDYKNTFPKWKPGSLASHVKNLDENGLDLLSKMLIYDPAKRISGKMALNHPYFNDLDNQIKKM DIP protein P06493 original sequence version: 185 DIP protein P06493 original sequence version: 162 DIP protein P06493 original sequence version: 184 DIP protein P06493 original sequence version: 97 DIP protein P06493 original sequence version: 192 942D79448EFE490A cip1_human E3 ubiquitin-protein ligase CCNB1IP1 Cyclin-B1-interacting protein 1 Human enhancer of invasion 10 CCNB1IP1 C14orf18 HEI10 RING-type E3 ubiquitin transferase CCNB1IP1 protein protein human Homo sapiens Human MSLCEDMLLCNYRKCRIKLSGYAWVTACSHIFCDQHGSGEFSRSPAICPACNSTLSGKLDIVRTELSPSEEYKAMVLAGLRPEIVLDISSRALAFWTYQVHQERLYQEYNFSKAEGHLKQMEKIYTQQIQSKDVELTSMKGEVTSMKKVLEEYKKKFSDISEKLMERNRQYQKLQGLYDSLRLRNITIANHEGTLEPSMIAQSGVLGFPLGNNSKFPLDNTPVRNRGDGDGDFQFRPFFAGSPTAPEPSNSFFSFVSPSRELEQQQVSSRAFKVKRI 885723B01A35225A cnb1ip1-ccnb1 1 4 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 8 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association the two-hybrid system was used to identify potential HEI10 interactive partners by using a bait representing the amino-terminal 143 aa of HEI10. Clones encoding cyclin B1 (aa 315 to end [5]) and the E2 ubiquitin-conjugating enzyme UbcH7 (aa 29 to end [41]) were isolated as candidate interacting partners of HEI10. cnb1ip1-ube2l3 1 8 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 5 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 physical association physical association the two-hybrid system was used to identify potential HEI10 interactive partners by using a bait representing the amino-terminal 143 aa of HEI10. Clones encoding cyclin B1 (aa 315 to end [5]) and the E2 ubiquitin-conjugating enzyme UbcH7 (aa 29 to end [41]) were isolated as candidate interacting partners of HEI10. cdc2-cip1 2 4 unspecified role unspecified role ancillary ancillary spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 7 enzyme enzyme unspecified role unspecified role 8 enzyme target enzyme target substrate substrate unspecified role unspecified role 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 region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain Q9NPC3:p.[Thr221Ala;Pro222Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain T A certain certain sequence position certain certain certain sequence position certain P A Wed May 02 19:00:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr221ala,pro222ala' Tue Jun 30 17:28:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Thr221Ala;Pro222Ala]' ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Purified bacterially expressed GST-HEI10 was incubated with purified active cyclin B-cdc2 complex in the presence of [-32P]ATP. The presence of an 65-kDa band corresponding to autophosphorylated cyclin B indicates that the cyclin B-cdc2 complex is active (Fig. 7A, lanes 1 to 4). When HEI10-GST was added to the reaction as a substrate, an additional phosphorylated band (60 kDa) that comigrates with HEI10-GST was observed 7 HEI10 is phosphorylated by cyclin B1-cdc2. Phosphorylation was abrogated in HEI10-Delta200-277, whereas it was strikingly reduced in upon mutation of TP221,222. cnb1ip1-ubiq 3 6 unspecified role unspecified role prey prey 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) physical association physical association 6 B GST-HEI10 incubated with noncycling Xenopus extracts developed an extensive ladder of higher migrating species. The modified forms were most abundant in interphase extracts, although they were observed in mitotic extracts as well. The periodicity of molecular masses observed approximated the 9-kDa ladder characteristic of ubiquitin modification; in fact, reprobing of lanes with antibody to ubiquitin (Fig. 6B) showed significant reactivity in lanes containing GST-HEI10 cnb1ip1-ubiq-1 3 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) 6 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) 5 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) association association 6 C GST-HEI10 is ubiquitinated in the presence of UbcH7 but not in the presence of either of the other E2 proteins.