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/ http://www.ebi.ac.uk/ makowska_grzyska-2010-3 Primase directs the release of DnaC from DnaB. in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Makowska-Grzyska M., Kaguni J.M. kaguni@msu.edu Mol. Cell (1097-2765) 2010 Only protein-protein interactions imex curation makowska_grzyska-2010-1 Primase directs the release of DnaC from DnaB. in vitro In vitro crosslink cross-linking study crosslink weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Makowska-Grzyska M., Kaguni J.M. kaguni@msu.edu Mol. Cell (1097-2765) 2010 Only protein-protein interactions imex curation makowska_grzyska-2010-2 Primase directs the release of DnaC from DnaB. in vitro In vitro filter binding filter binding Filter overlay assay Filter overlay assay predetermined predetermined participant predetermined Makowska-Grzyska M., Kaguni J.M. kaguni@msu.edu Mol. Cell (1097-2765) 2010 Only protein-protein interactions imex curation makowska_grzyska-2010-4 Primase directs the release of DnaC from DnaB. in vitro In vitro comigration in sds comigration in sds page comigration in sds western blot western blot Immuno blot Immuno blot Makowska-Grzyska M., Kaguni J.M. kaguni@msu.edu Mol. Cell (1097-2765) 2010 Only protein-protein interactions imex curation dnac_ecoli DNA replication protein DnaC dnaC dnaD b4361 JW4325 protein protein ecoli Escherichia coli (strain K12) MKNVGDLMQRLQKMMPAHIKPAFKTGEELLAWQKEQGAIRSAALERENRAMKMQRTFNRSGIRPLHQNCSFENYRVECEGQMNALSKARQYVEEFDGNIASFIFSGKPGTGKNHLAAAICNELLLRGKSVLIITVADIMSAMKDTFRNSGTSEEQLLNDLSNVDLLVIDEIGVQTESKYEKVIINQIVDRRSSSKRPTGMLTNSNMEEMTKLLGERVMDRMRLGNSLWVIFNWDSYRSRVTGKEY atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) ssdna single-stranded dna ss dna single stranded deoxyribonucleic acid ss DNA ss dna chemical synthesis Chemical synthesis (Chemical synthesis) CATCCAATAAATCATACACAAGGC dnab_ecoli Replicative DNA helicase dnaB groP grpA b4052 JW4012 protein protein ecoli Escherichia coli (strain K12) MAGNKPFNKQQAEPRERDPQVAGLKVPPHSIEAEQSVLGGLMLDNERWDDVAERVVADDFYTRPHRHIFTEMARLQESGSPIDLITLAESLERQGQLDSVGGFAYLAELSKNTPSAANISAYADIVRERAVVREMISVANEIAEAGFDPQGRTSEDLLDLAESRVFKIAESRANKDEGPKNIADVLDATVARIEQLFQQPHDGVTGVNTGYDDLNKKTAGLQPSDLIIVAARPSMGKTTFAMNLVENAAMLQDKPVLIFSLEMPSEQIMMRSLASLSRVDQTKIRTGQLDDEDWARISGTMGILLEKRNIYIDDSSGLTPTEVRSRARRIAREHGGIGLIMIDYLQLMRVPALSDNRTLEIAEISRSLKALAKELNVPVVALSQLNRSLEQRADKRPVNSDLRESGSIEQDADLIMFIYRDEVYHENSDLKGIAEIIIGKQRNGPIGTVRLTFNGQWSRFDNYAGPQYDDE m13oric2lb5 M13oriC2LB5 supercoiled DNA plasmid containing E.coli replication origin dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid chemical synthesis Chemical synthesis (Chemical synthesis) dnaa_ecoli Chromosomal replication initiator protein DnaA dnaA b3702 JW3679 protein protein ecoli Escherichia coli (strain K12) MSLSLWQQCLARLQDELPATEFSMWIRPLQAELSDNTLALYAPNRFVLDWVRDKYLNNINGLLTSFCGADAPQLRFEVGTKPVTQTPQAAVTSNVAAPAQVAQTQPQRAAPSTRSGWDNVPAPAEPTYRSNVNVKHTFDNFVEGKSNQLARAAARQVADNPGGAYNPLFLYGGTGLGKTHLLHAVGNGIMARKPNAKVVYMHSERFVQDMVKALQNNAIEEFKRYYRSVDALLIDDIQFFANKERSQEEFFHTFNALLEGNQQIILTSDRYPKEINGVEDRLKSRFGWGLTVAIEPPELETRVAILMKKADENDIRLPGEVAFFIAKRLRSNVRELEGALNRVIANANFTGRAITIDFVREALRDLLALQEKLVTIDNIQKTVAEYYKIKVADLLSKRRSRSVARPRQMAMALAKELTNHSLPEIGDAFGGRDHTTVLHACRKIEQLREESHDIKEDFSNLIRTLSS dnac-ssdna-atp 1362995 n/a 1362999 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) n/a 1363000 unspecified role unspecified role prey prey chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1363001 unspecified role unspecified role bait bait 5-prime biotin tag biotin tag certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association false false false 1362995 Fig. 3 ssdna-dnac 1362995 n/a 1362999 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) n/a 1363001 unspecified role unspecified role bait bait 5-prime biotin tag biotin tag certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false 1362995 Fig. 3 dnac-atp 1362996 n/a 1363000 unspecified role unspecified role unspecified role unspecified role region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1362999 unspecified role unspecified role unspecified role unspecified role ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig. 2a dnab-dnac-1 1362997 n/a 1363002 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) n/a 1362999 unspecified role unspecified role prey prey P0AEF0:p.Leu11Gln mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig. 2b, c dnaa-dnab-dnac 1362998 n/a 1363003 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 n/a 1363004 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 9.0 n/a 1363002 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 11.0 n/a 1362999 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 6.0 physical association physical association false false false Fig. 5, 6 Stoichiometry of DnaA, DnaB and DnaC (wildtype) calculated relative to one copy of the oriC plasmid in an assay containing M13oriC2LB5, DnaA, DnaB, DnaC, HU, SSB and ATP/ATPgammaS. Stoichiometry changes slightly depending on which prepriming and replication complex enzymes were added to the assay (Fig. 6). Binding of DnaC lost if DnaG (primase) and rNTPs added.