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 interaction of DiaA and DnaA regulates the replication cycle in E. coli by directly promoting ATP DnaA-specific initiation complexes. The interaction of DiaA and DnaA regulates the replication cycle in E. coli by directly promoting ATP DnaA-specific initiation complexes. Genes Dev. (0890-9369) 2007 imex curation Keyamura K., Fujikawa N., Ishida T., Ozaki S., Su'etsugu M., Fujimitsu K., Kagawa W., Yokoyama S., Kurumizaka H., Katayama T. katayama@phar.kyushu-u.ac.jp,kurumizaka@waseda.jp 27-Mar-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip predetermined predetermined participant predetermined Keyamura K., Fujikawa N., Ishida T., Ozaki S., Su'etsugu M., Fujimitsu K., Kagawa W., Yokoyama S., Kurumizaka H., Katayama T. katayama@phar.kyushu-u.ac.jp,kurumizaka@waseda.jp Accepted 2013-FEB-28 by ORCHARD 27-Mar-2013: Contacted by IntAct-Help. Genes Dev. (0890-9369) 2007 Only protein-protein interactions imex curation The interaction of DiaA and DnaA regulates the replication cycle in E. coli by directly promoting ATP DnaA-specific initiation complexes. The interaction of DiaA and DnaA regulates the replication cycle in E. coli by directly promoting ATP DnaA-specific initiation complexes. Genes Dev. (0890-9369) 2007 imex curation Keyamura K., Fujikawa N., Ishida T., Ozaki S., Su'etsugu M., Fujimitsu K., Kagawa W., Yokoyama S., Kurumizaka H., Katayama T. katayama@phar.kyushu-u.ac.jp,kurumizaka@waseda.jp 27-Mar-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Keyamura K., Fujikawa N., Ishida T., Ozaki S., Su'etsugu M., Fujimitsu K., Kagawa W., Yokoyama S., Kurumizaka H., Katayama T. katayama@phar.kyushu-u.ac.jp,kurumizaka@waseda.jp Accepted 2013-FEB-28 by ORCHARD 27-Mar-2013: Contacted by IntAct-Help. Genes Dev. (0890-9369) 2007 Only protein-protein interactions imex curation The interaction of DiaA and DnaA regulates the replication cycle in E. coli by directly promoting ATP DnaA-specific initiation complexes. The interaction of DiaA and DnaA regulates the replication cycle in E. coli by directly promoting ATP DnaA-specific initiation complexes. Genes Dev. (0890-9369) 2007 imex curation Keyamura K., Fujikawa N., Ishida T., Ozaki S., Su'etsugu M., Fujimitsu K., Kagawa W., Yokoyama S., Kurumizaka H., Katayama T. katayama@phar.kyushu-u.ac.jp,kurumizaka@waseda.jp 27-Mar-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight by comassie molecular weight estimation by coomasie staining weight by comassie Keyamura K., Fujikawa N., Ishida T., Ozaki S., Su'etsugu M., Fujimitsu K., Kagawa W., Yokoyama S., Kurumizaka H., Katayama T. katayama@phar.kyushu-u.ac.jp,kurumizaka@waseda.jp Accepted 2013-FEB-28 by ORCHARD 27-Mar-2013: Contacted by IntAct-Help. Genes Dev. (0890-9369) 2007 Only protein-protein interactions imex curation The interaction of DiaA and DnaA regulates the replication cycle in E. coli by directly promoting ATP DnaA-specific initiation complexes. The interaction of DiaA and DnaA regulates the replication cycle in E. coli by directly promoting ATP DnaA-specific initiation complexes. Genes Dev. (0890-9369) 2007 imex curation Keyamura K., Fujikawa N., Ishida T., Ozaki S., Su'etsugu M., Fujimitsu K., Kagawa W., Yokoyama S., Kurumizaka H., Katayama T. katayama@phar.kyushu-u.ac.jp,kurumizaka@waseda.jp 27-Mar-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro density sedimentation cosedimentation through density gradient density sedimentation weight by comassie molecular weight estimation by coomasie staining weight by comassie Keyamura K., Fujikawa N., Ishida T., Ozaki S., Su'etsugu M., Fujimitsu K., Kagawa W., Yokoyama S., Kurumizaka H., Katayama T. katayama@phar.kyushu-u.ac.jp,kurumizaka@waseda.jp Accepted 2013-FEB-28 by ORCHARD 27-Mar-2013: Contacted by IntAct-Help. Genes Dev. (0890-9369) 2007 Only protein-protein interactions imex curation dnaa_ecoli Chromosomal replication initiator protein DnaA dnaA b3702 JW3679 protein protein ecoli Escherichia coli (strain K12) MSLSLWQQCLARLQDELPATEFSMWIRPLQAELSDNTLALYAPNRFVLDWVRDKYLNNINGLLTSFCGADAPQLRFEVGTKPVTQTPQAAVTSNVAAPAQVAQTQPQRAAPSTRSGWDNVPAPAEPTYRSNVNVKHTFDNFVEGKSNQLARAAARQVADNPGGAYNPLFLYGGTGLGKTHLLHAVGNGIMARKPNAKVVYMHSERFVQDMVKALQNNAIEEFKRYYRSVDALLIDDIQFFANKERSQEEFFHTFNALLEGNQQIILTSDRYPKEINGVEDRLKSRFGWGLTVAIEPPELETRVAILMKKADENDIRLPGEVAFFIAKRLRSNVRELEGALNRVIANANFTGRAITIDFVREALRDLLALQEKLVTIDNIQKTVAEYYKIKVADLLSKRRSRSVARPRQMAMALAKELTNHSLPEIGDAFGGRDHTTVLHACRKIEQLREESHDIKEDFSNLIRTLSS DIP protein P03004 original sequence version: 109 DIP protein P03004 original sequence version: null 607C8366A8CDCCED diaa_ecoli DnaA initiator-associating protein DiaA diaA b3149 JW3118 yraO protein protein ecoli Escherichia coli (strain K12) MQERIKACFTESIQTQIAAAEALPDAISRAAMTLVQSLLNGNKILCCGNGTSAANAQHFAASMINRFETERPSLPAIALNTDNVVLTAIANDRLHDEVYAKQVRALGHAGDVLLAISTRGNSRDIVKAVEAAVTRDMTIVALTGYDGGELAGLLGPQDVEIRIPSHRSARIQEMHMLTVNCLCDLIDNTLFPHQDD F9ADFE9C0DF365A8 diaa homotetramer 2 6 unspecified role unspecified role neutral component neutral component 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 1b, c, S1 Rwork = 0.18; Rfree = 0.21 1.85A diaa homomultimer 3 6 unspecified role unspecified role neutral component neutral component 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 ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig.s 3a diaa homodimer mut 3 6 unspecified role unspecified role neutral component neutral component P66817:p.Lys101Glu mutation mutation certain certain sequence position certain certain certain sequence position certain K E Wed May 02 19:00:16 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys101glu' Tue Jun 30 19:07:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys101Glu' P66817:p.Asn83Asp mutation mutation certain certain sequence position certain certain certain sequence position certain N D Wed May 02 19:42:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn83asp' Tue Jun 30 17:24:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn83Asp' 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig.s 3a diaa homotetramer 4 6 unspecified role unspecified role neutral component neutral component 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig.s 3c diaa homodimer mut 4 6 unspecified role unspecified role neutral component neutral component 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 P66817:p.Lys101Glu mutation mutation certain certain sequence position certain certain certain sequence position certain K E Wed May 02 20:21:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys101glu' Tue Jun 30 17:44:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys101Glu' P66817:p.Asn83Asp mutation mutation certain certain sequence position certain certain certain sequence position certain N D Wed May 02 18:08:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn83asp' Tue Jun 30 16:17:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn83Asp' ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig.s 3c dnaa-diaa 1 6 unspecified role unspecified role prey prey P66817:p.Phe191Ser mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain F S Wed May 02 17:22:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe191ser' Tue Jun 30 17:54:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe191Ser' P66817:p.Phe191Leu mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain F L Wed May 02 15:49:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe191leu' Tue Jun 30 15:58:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe191Leu' 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 P66817:p.Leu190Pro mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain L P Wed May 02 14:38:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu190pro' Tue Jun 30 16:56:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu190Pro' P66817:p.Asn83Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain N D Wed May 02 17:30:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn83asp' Tue Jun 30 16:52:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn83Asp' P66817:p.Pro72Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain P A Wed May 02 15:01:06 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro72ala' Tue Jun 30 17:49:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro72Ala' P66817:p.Pro72Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain P S Wed May 02 14:41:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro72ser' Tue Jun 30 17:45:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro72Ser' P66817:p.Leu190Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain L A Wed May 02 16:36:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu190ala' Tue Jun 30 17:37:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu190Ala' P66817:p.Lys101Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K E Wed May 02 15:15:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys101glu' Tue Jun 30 20:10:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys101Glu' ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role bait bait n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 5 dnaa-diaa-1 1 6 unspecified role unspecified role prey prey P66817:p.Asn83Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain N D Wed May 02 15:35:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn83asp' Tue Jun 30 19:22:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn83Asp' 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 P66817:p.Leu190Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain L A Wed May 02 19:34:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu190ala' Tue Jun 30 20:21:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu190Ala' P66817:p.Ser52Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain S F Wed May 02 15:25:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser52phe' Tue Jun 30 18:07:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser52Phe' ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role bait bait n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 6a This is a 2-stage anti-tag coip using a restricted amount of biotin-tagged DnaA in the first pulldown and excess amounts of untagged DnaA in the second pulldown before visualising the complex on SDS-PAGE. This allowed for several different DnaA molecules to be pulled down by one DiaA homotetramer. diaa-dnaa 1 5 unspecified role unspecified role prey prey n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 6 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 6b