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/ ATP-dependent assembly of the human origin recognition complex. ATP-dependent assembly of the human origin recognition complex. J. Biol. Chem. (0021-9258) 2007 imex curation Siddiqui K., Stillman B. stillman@cshl.edu 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Siddiqui K., Stillman B. stillman@cshl.edu Accepted 2012-JUL-19 by JYOTI 19-Sept-2012: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation ATP-dependent assembly of the human origin recognition complex. ATP-dependent assembly of the human origin recognition complex. J. Biol. Chem. (0021-9258) 2007 imex curation Siddiqui K., Stillman B. stillman@cshl.edu 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. pull down pull down weight silver stain molecular weight estimation by silver staining weight silver stain Siddiqui K., Stillman B. stillman@cshl.edu Accepted 2012-JUL-19 by JYOTI 19-Sept-2012: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation ATP-dependent assembly of the human origin recognition complex. ATP-dependent assembly of the human origin recognition complex. J. Biol. Chem. (0021-9258) 2007 imex curation Siddiqui K., Stillman B. stillman@cshl.edu 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Siddiqui K., Stillman B. stillman@cshl.edu Accepted 2012-JUL-19 by JYOTI 19-Sept-2012: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation ATP-dependent assembly of the human origin recognition complex. ATP-dependent assembly of the human origin recognition complex. J. Biol. Chem. (0021-9258) 2007 imex curation Siddiqui K., Stillman B. stillman@cshl.edu 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Siddiqui K., Stillman B. stillman@cshl.edu Accepted 2012-JUL-19 by JYOTI 19-Sept-2012: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation ATP-dependent assembly of the human origin recognition complex. ATP-dependent assembly of the human origin recognition complex. J. Biol. Chem. (0021-9258) 2007 imex curation Siddiqui K., Stillman B. stillman@cshl.edu 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Siddiqui K., Stillman B. stillman@cshl.edu Accepted 2012-JUL-19 by JYOTI 19-Sept-2012: Contacted by IntAct-Help. Time after G1/S transition arrest (hours) J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation orc6_human Origin recognition complex subunit 6 ORC6 ORC6L protein protein human Homo sapiens Human MGSELIGRLAPRLGLAEPDMLRKAEEYLRLSRVKCVGLSARTTETSSAVMCLDLAASWMKCPLDRAYLIKLSGLNKETYQSCLKSFECLLGLNSNIGIRDLAVQFSCIEAVNMASKILKSYESSLPQTQQVDLDLSRPLFTSAALLSACKILKLKVDKNKMVATSGVKKAIFDRLCKQLEKIGQQVDREPGDVATPPRKRKKIVVEAPAKEMEKVEEMPHKPQKDEDLTQDYEEWKRKILENAASAQKATAE DIP protein Q9Y5N6 original sequence version: 119 DIP protein Q9Y5N6 original sequence version: 56 DIP protein Q9Y5N6 original sequence version: 130 78840387605F45FE orc2_human Origin recognition complex subunit 2 ORC2 ORC2L protein protein human Homo sapiens Human MSKPELKEDKMLEVHFVGDDDVLNHILDREGGAKLKKERAQLLVNPKKIIKKPEYDLEEDDQEVLKDQNYVEIMGRDVQESLKNGSATGGGNKVYSFQNRKHSEKMAKLASELAKTPQKSVSFSLKNDPEITINVPQSSKGHSASDKVQPKNNDKSEFLSTAPRSLRKRLIVPRSHSDSESEYSASNSEDDEGVAQEHEEDTNAVIFSQKIQAQNRVVSAPVGKETPSKRMKRDKTSDLVEEYFEAHSSSKVLTSDRTLQKLKRAKLDQQTLRNLLSKVSPSFSAELKQLNQQYEKLFHKWMLQLHLGFNIVLYGLGSKRDLLERFRTTMLQDSIHVVINGFFPGISVKSVLNSITEEVLDHMGTFRSILDQLDWIVNKFKEDSSLELFLLIHNLDSQMLRGEKSQQIIGQLSSLHNIYLIASIDHLNAPLMWDHAKQSLFNWLWYETTTYSPYTEETSYENSLLVKQSGSLPLSSLTHVLRSLTPNARGIFRLLIKYQLDNQDNPSYIGLSFQDFYQQCREAFLVNSDLTLRAQLTEFRDHKLIRTKKGTDGVEYLLIPVDNGTLTDFLEKEEEEA DIP protein Q13416 original sequence version: 117 DIP protein Q13416 original sequence version: 56 DIP protein Q13416 original sequence version: 94 DF3F9C2CF147DA5F orc3_human Origin recognition complex subunit 3 ORC3 LATHEO Origin recognition complex subunit Latheo ORC3L protein protein human Homo sapiens Human MATSSMSKGCFVFKPNSKKRKISLPIEDYFNKGKNEPEDSKLRFETYQLIWQQMKSENERLQEELNKNLFDNLIEFLQKSHSGFQKNSRDLGGQIKLREIPTAALVLGVNVTDHDLTFGSLTEALQNNVTPYVVSLQAKDCPDMKHFLQKLISQLMDCCVDIKSKEEESVHVTQRKTHYSMDSLSSWYMTVTQKTDPKMLSKKRTTSSQWQSPPVVVILKDMESFATKVLQDFIIISSQHLHEFPLILIFGIATSPIIIHRLLPHAVSSLLCIELFQSLSCKEHLTTVLDKLLLTTQFPFKINEKVLQVLTNIFLYHDFSVQNFIKGLQLSLLEHFYSQPLSVLCCNLPEAKRRINFLSNNQCENIRRLPSFRRYVEKQASEKQVALLTNERYLKEETQLLLENLHVYHMNYFLVLRCLHKFTSSLPKYPLGRQIRELYCTCLEKNIWDSEEYASVLQLLRMLAKDELMTILEKCFKVFKSYCENHLGSTAKRIEEFLAQFQSLDETKEEEDASGSQPKGLQKTDLYHLQKSLLEMKELRRSKKQTKFEVLRENVVNFIDCLVREYLLPPETQPLHEVVYFSAAHALREHLNAAPRIALHTALNNPYYYLKNEALKSEEGCIPNIAPDICIAYKLHLECSRLINLVDWSEAFATVVTAAEKMDANSATSEEMNEIIHARFIRAVSELELLGFIKPTKQKTDHVARLTWGGC DIP protein Q9UBD5 original sequence version: 124 DIP protein Q9UBD5-1 original sequence version: 96 DIP protein Q9UBD5 original sequence version: 100 E713569A11B39B1C orc1_human Origin recognition complex subunit 1 ORC1 ORC1L PARC1 Replication control protein 1 protein protein human Homo sapiens Human MAHYPTRLKTRKTYSWVGRPLLDRKLHYQTYREMCVKTEGCSTEIHIQIGQFVLIEGDDDENPYVAKLLELFEDDSDPPPKKRARVQWFVRFCEVPACKRHLLGRKPGAQEIFWYDYPACDSNINAETIIGLVRVIPLAPKDVVPTNLKNEKTLFVKLSWNEKKFRPLSSELFAELNKPQESAAKCQKPVRAKSKSAESPSWTPAEHVAKRIESRHSASKSRQTPTHPLTPRARKRLELGNLGNPQMSQQTSCASLDSPGRIKRKVAFSEITSPSKRSQPDKLQTLSPALKAPEKTRETGLSYTEDDKKASPEHRIILRTRIAASKTIDIREERTLTPISGGQRSSVVPSVILKPENIKKRDAKEAKAQNEATSTPHRIRRKSSVLTMNRIRQQLRFLGNSKSDQEEKEILPAAEISDSSSDEEEASTPPLPRRAPRTVSRNLRSSLKSSLHTLTKVPKKSLKPRTPRCAAPQIRSRSLAAQEPASVLEEARLRLHVSAVPESLPCREQEFQDIYNFVESKLLDHTGGCMYISGVPGTGKTATVHEVIRCLQQAAQANDVPPFQYIEVNGMKLTEPHQVYVQILQKLTGQKATANHAAELLAKQFCTRGSPQETTVLLVDELDLLWTHKQDIMYNLFDWPTHKEARLVVLAIANTMDLPERIMMNRVSSRLGLTRMCFQPYTYSQLQQILRSRLKHLKAFEDDAIQLVARKVAALSGDARRCLDICRRATEICEFSQQKPDSPGLVTIAHSMEAVDEMFSSSYITAIKNSSVLEQSFLRAILAEFRRSGLEEATFQQIYSQHVALCRMEGLPYPTMSETMAVCSHLGSCRLLLVEPSRNDLLLRVRLNVSQDDVLYALKDE DIP protein Q13415 original sequence version: 131 DIP protein Q13415 original sequence version: 64 DIP protein Q13415 original sequence version: 100 DIP protein Q13415 original sequence version: 105 5C594553F7F808E2 orc5_human Origin recognition complex subunit 5 ORC5 ORC5L protein protein human Homo sapiens Human MPHLENVVLCRESQVSILQSLFGERHHFSFPSIFIYGHTASGKTYVTQTLLKTLELPHVFVNCVECFTLRLLLEQILNKLNHLSSSEDGCSTEITCETFNDFVRLFKQVTTAENLKDQTVYIVLDKAEYLRDMEANLLPGFLRLQELADRNVTVLFLSEIVWEKFRPNTGCFEPFVLYFPDYSIGNLQKILSHDHPPEYSADFYAAYINILLGVFYTVCRDLKELRHLAVLNFPKYCEPVVKGEASERDTRKLWRNIEPHLKKAMQTVYLREISSSQWEKLQKDDTDPGQLKGLSAHTHVELPYYSKFILIAAYLASYNPARTDKRFFLKHHGKIKKTNFLKKHEKTSNHLLGPKPFPLDRLLAILYSIVDSRVAPTANIFSQITSLVTLQLLTLVGHDDQLDGPKYKCTVSLDFIRAIARTVNFDIIKYLYDFL DIP protein O43913 original sequence version: 116 DIP protein O43913 original sequence version: 53 DIP protein O43913 original sequence version: 90 0FEB0E829C124745 orc4_human Origin recognition complex subunit 4 ORC4 ORC4L protein protein human Homo sapiens Human MSSRKSKSNSLIHTECLSQVQRILRERFCRQSPHSNLFGVQVQYKHLSELLKRTALHGESNSVLIIGPRGSGKTMLINHALKELMEIEEVSENVLQVHLNGLLQINDKIALKEITRQLNLENVVGDKVFGSFAENLSFLLEALKKGDRTSSCPVIFILDEFDLFAHHKNQTLLYNLFDISQSAQTPIAVIGLTCRLDILELLEKRVKSRFSHRQIHLMNSFGFPQYVKIFKEQLSLPAEFPDKVFAEKWNENVQYLSEDRSVQEVLQKHFNISKNLRSLHMLLMLALNRVTASHPFMTAVDLMEASQLCSMDSKANIVHGLSVLEICLIIAMKHLNDIYEEEPFNFQMVYNEFQKFVQRKAHSVYNFEKPVVMKAFEHLQQLELIKPMERTSGNSQREYQLMKLLLDNTQIMNALQKYPNCPTDVRQWATSSLSWL DIP protein O43929 original sequence version: 134 DIP protein O43929 original sequence version: 65 A7020B6690E30B4E orc2-orc3-2 2 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range association association Fig. 1A, 1B ATP ( or a a nucleoside triphosphate analogue) is required for the formation of a stable Orc2/Orc3/Orc4/Orc5 complex. orc2-orc3-3 4 10 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 region t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 7 unspecified role unspecified role bait bait n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 8 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 rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 11 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes association association Fig. 1C, 2C ATP ( or a a nucleoside triphosphate analogue) is required for the formation of a stable Orc2/Orc3/Orc4/Orc5 complex. orc2-orc3-7 4 8 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 rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 7 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 11 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes association association Fig. 2B ORC4 Lys73Ala, Lys73Glu, or Asp_Glu159-160Ala_Ala mutations which stop ORC4 binding ATP also prevent ORC4 binding to this complex. orc2-orc3-5 2 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role prey prey association association Fig. 3A This sub-complex will assemble in the absence of ATP orc2-orc1-1 2 8 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 9 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range association association Fig. 3A ATP ( or a a nucleoside triphosphate analogue) is required for the formation of a stable Orc2/Orc3/Orc4/Orc5 complex. orc1-orc2-2 4 11 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 8 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 rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 9 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role prey prey region t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 7 unspecified role unspecified role prey prey n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes association association Fig. 3C, 4A, 4B, 4C ATP is required for the formation of a stable Orc1/Orc2/Orc3/Orc4/Orc5 complex, however mutations in Orc1 predicted to disrupt ATP binding or hydrolysis have no significant effect in complex formation in vitro. ATP binding by Orc1 appears not to be essential for complex formation and may have other roles in DNA replication. Association of Orc1 with Orc2/Orc3/Orc5 is Orc4-dependent and requires a functional ATP-binding site in Orc4. orc1-orc4 4 11 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 10 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 rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 9 unspecified role unspecified role bait bait n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) association association Fig. 4C orc1-orc5 4 9 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 10 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 region t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes physical association physical association Fig. 4C orc1-orc2-3 3 7 unspecified role unspecified role prey prey 9 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 region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 10 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey association association Fig. 5B orc1-orc2-4 1 8 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey association association Fig.5B orc1-orc2-5 5 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey association association Fig.6B 0 orc3-orc1 5 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey association association Fig.6B 1 orc3-orc1-1 5 11 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey association association Fig.6B 2 orc3-orc1-2 5 11 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey association association Fig.6B 3 orc3-orc1-3 5 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait association association Fig.6B 4 orc3-orc1-4 5 10 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey association association Fig.6B 5 orc3-orc1-5 5 11 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey association association Fig.6B 6 orc3-orc1-6 5 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait association association Fig.6B 7 orc3-orc2-2 5 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait association association Fig.6B 8 orc3-orc2-3 5 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait association association Fig.6B 9 orc3-orc2-4 5 8 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey association association Fig.6B 10 orc3-orc1-7 1 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait association association Fig.7A orc4-orc2 1 7 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey association association Fig.7A orc4-orc5-2 1 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait association association Fig.7A orc3-orc1-8 1 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey association association Fig.7B Low levels of ORC6 was seen in the interaction suggesting that a large fraction of Orc6 was not associated with the ORC complex. orc6-orc3-1 4 8 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 trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 6 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 7C orc2-orc3-6 4 7 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 10 unspecified role unspecified role prey prey region t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 8 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 rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes association association Fig. 2C ORC5 Lys43Ala and Asp-Lys125-126Ala-Ala mutations which prevent ORC5 binding ATP prevent ORC5 binding to this complex.