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/ paul-2008-2 Allosteric regulation of histidine kinases by their cognate response regulator determines cell fate. in vitro In vitro phosphotransferase phosphotransferase assay phosphotransferase weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Paul R., Jaeger T., Abel S., Wiederkehr I., Folcher M., Biondi EG., Laub MT., Jenal U. Diguanylate cyclase assay catalyzes the condensation of two GTP molecules to the cyclic dinucleotide di-GMP (c-di-GMP), which acts as a secondary messenger. urs.jenal@unibas.ch Cell (0092-8674) 2008 Only protein-protein interactions 18 imex curation paul-2008-6 Allosteric regulation of histidine kinases by their cognate response regulator determines cell fate. in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Paul R., Jaeger T., Abel S., Wiederkehr I., Folcher M., Biondi EG., Laub MT., Jenal U. Diguanylate cyclase assay catalyzes the condensation of two GTP molecules to the cyclic dinucleotide di-GMP (c-di-GMP), which acts as a secondary messenger. urs.jenal@unibas.ch Cell (0092-8674) 2008 Only protein-protein interactions 18 imex curation paul-2008-3 Allosteric regulation of histidine kinases by their cognate response regulator determines cell fate. in vitro In vitro phosphatase assay phosphatase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Paul R., Jaeger T., Abel S., Wiederkehr I., Folcher M., Biondi EG., Laub MT., Jenal U. Diguanylate cyclase assay catalyzes the condensation of two GTP molecules to the cyclic dinucleotide di-GMP (c-di-GMP), which acts as a secondary messenger. urs.jenal@unibas.ch Cell (0092-8674) 2008 Only protein-protein interactions 18 imex curation paul-2008-4 Allosteric regulation of histidine kinases by their cognate response regulator determines cell fate. in vitro In vitro crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Paul R., Jaeger T., Abel S., Wiederkehr I., Folcher M., Biondi EG., Laub MT., Jenal U. Diguanylate cyclase assay catalyzes the condensation of two GTP molecules to the cyclic dinucleotide di-GMP (c-di-GMP), which acts as a secondary messenger. Purified protein was cross-linked using 2 mM disuccinimidyl suberate. urs.jenal@unibas.ch Cell (0092-8674) 2008 Only protein-protein interactions 18 imex curation paul-2008-5 Allosteric regulation of histidine kinases by their cognate response regulator determines cell fate. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight identificat confirmation by molecular weight weight identificat Paul R., Jaeger T., Abel S., Wiederkehr I., Folcher M., Biondi EG., Laub MT., Jenal U. Diguanylate cyclase assay catalyzes the condensation of two GTP molecules to the cyclic dinucleotide di-GMP (c-di-GMP), which acts as a secondary messenger. Multi-angle light scattering was used to measure the molecular weight of purified proteins. urs.jenal@unibas.ch Cell (0092-8674) 2008 Only protein-protein interactions 18 imex curation paul-2008-1 Allosteric regulation of histidine kinases by their cognate response regulator determines cell fate. in vitro In vitro enzymatic study enzymatic study predetermined predetermined participant predetermined Paul R., Jaeger T., Abel S., Wiederkehr I., Folcher M., Biondi EG., Laub MT., Jenal U. Diguanylate cyclase assay catalyzes the condensation of two GTP molecules to the cyclic dinucleotide di-GMP (c-di-GMP), which acts as a secondary messenger. urs.jenal@unibas.ch Cell (0092-8674) 2008 Only protein-protein interactions 18 imex curation plec_cauvc Non-motile and phage-resistance protein pleC CC_2482 protein protein caucr Caulobacter crescentus (strain ATCC 19089 / CB15) Caulobacter vibrioides MGRHGGPAAAGPTAPSAVRAKAVNAPSQVFVRIAILAALLLLAVYTAFGVHRLQREAMAQPGGAPLAAKADLIAGRVDANLAAQRAGLSAAADLLKRDPGATMDAAETTLRAAGGEAAAVAVVSEAGVVAVAGRDDGADWKAAALAAGASGRTNWVGSVGETGRLYVATTTSLDRARAFVIASGDASRLVADPEKGESGALALPDGKLIAARGRGVQGAGALREAFALSIEDLGDGPAAVRGQAADGALLDVAVRPVAQGALLAVAAAPTRSVANLDRQVMEGAFSLLVPLGVGIALALLLMIQSRKAEVAHREFIDSERRFRLAVEAARCGIWEWDLNGDQVYLSDVTGAMFGWGGGGVVSGQDLLERISIDHRERVRQALANAAMYGAFDVSFRVPASEQGARSLWIDARGQGFGKPGSEGHARIIGVALDVTEERIAQARAQAAENRLRDAIESVSEAFVLWDRQGRLLMCNRNYRSVFSLEPKILKPGAARAEVNRFAALAIKQDHPAPDGAKGVREAEMMDGRWIQISERRTAEGGLVMTAADITAIKTQEEARRRNEEQLQNAVAGLERSQEQLAELARKYETEKVKAESANKAKSEFLANMSHELRTPLNAINGFSEIMMNEMFGPLGDQRYKGYSQDIHSSGQHLLALINDILDMSKIEAGKMNLKFESMHLEDVAEDAVRLVRNRAEAAGLKLDIDFPQLPEIEADYRAVKQVLLNLLSNAIKFTPRAGSVTVRAEVRRDPFGDLIKVSVTDTGIGIAKEDLARLAKPFEQVESQFSKTTQGTGLGLALTKSLITMHDGVLEMHSTPGEGTTVSFTLPVRHSDQKITRDFVAA DCFF4F092DE2CBF8 q45976_cauvi divK protein protein cauce Caulobacter crescentus Caulobacter crescentus MTKKVLIVEDNELNMKLFHDLLEAQGYETLQTREGLSALSIARENKPDLILMDIQLPEISGLEVTKWLKEDDDLAHIPVVAVTAFAMKGDEERIREGGCEAYISKPISVVHFLETIKRLLERQPA pled_cauvc Response regulator PleD pleD CC_2462 Stalked cell differentiation-controlling protein protein protein caucr Caulobacter crescentus (strain ATCC 19089 / CB15) Caulobacter vibrioides MSARILVVDDIEANVRLLEAKLTAEYYEVSTAMDGPTALAMAARDLPDIILLDVMMPGMDGFTVCRKLKDDPTTRHIPVVLITALDGRGDRIQGLESGASDFLTKPIDDVMLFARVRSLTRFKLVIDELRQREASGRRMGVIAGAAARLDGLGGRVLIVDDNERQAQRVAAELGVEHRPVIESDPEKAKISAGGPVDLVIVNAAAKNFDGLRFTAALRSEERTRQLPVLAMVDPDDRGRMVKALEIGVNDILSRPIDPQELSARVKTQIQRKRYTDYLRNNLDHSLELAVTDQLTGLHNRRYMTGQLDSLVKRATLGGDPVSALLIDIDFFKKINDTFGHDIGDEVLREFALRLASNVRAIDLPCRYGGEEFVVIMPDTALADALRIAERIRMHVSGSPFTVAHGREMLNVTISIGVSATAGEGDTPEALLKRADEGVYQAKASGRNAVVGKAA D44909D42B581516 divj_cauvc Histidine protein kinase DivJ divJ CC_1063 protein protein caucr Caulobacter crescentus (strain ATCC 19089 / CB15) Caulobacter vibrioides MILPTALKSRLALEFETLPDPFRRPAARAAGLDPAHAWRLGWLAAVCLAAAAALFTADSGGWPVWAALGAGALPALVSLIFTREDERTQSWLLVLWAVGGSLAAVLTGGVGGAMAAWCLAPVAAASTQDQPKRLAEGAALALIGACVAALTQLSGLAPAAPTGPLAFVLGFLALVTTGLGLAAGLLIGRRRQGARDDRYASEIIGLETLLDGLPHLAIAVRGQGQVTAVRGAAPPGVTRADLVNRGLTGAAAPGDRQRLTAAIAQAHREGSASLTFNPALGVERVVALDMHRVAPNQLVGVLRDITVERHREHALDQARIDAEALAAGRARFLANMSHELRTPLNAIMGFSDIMRARMFGPLSDRYAEYAELIHESGGHLLDLINDVLDMSKIEAERFELQRGVFDAREAVQAAMRLLRVQSDTAGVQLRGVLPPGELEVDADRRALKQIVLNLVSNALKFTPRGGQVTVTAHGYDGVLEIVVADTGVGISPEDLERLGRPYEQAGGAEQRARGTGLGLSLVRAFAQLHGGEMVIESRLGAGTTVSVRLPVLLAPMVAATPTPPAAPEAPSAPEPAPTVEEPPPASLGDNVIAFAPR 802B532B749B37EA gtp guanosine triphosphate - GTP 5'-GTP GTP Guanosine 5'-triphosphate guanosine 5'-triphosphoric acid guanosine triphosphate GUANOSINE-5'-TRIPHOSPHATE H4gtp guanosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) InChI=1/C10H16N5O14P3/c11-10-13-7-4(8(18)14-10)12-2-15(7)9-6(17)5(16)3(27-9)1-26-31(22,23)29-32(24,25)28-30(19,20)21/h2-3,5-6,9,16-17H,1H2,(H,22,23)(H,24,25)(H2,19,20,21)(H3,11,13,14,18)/t3-,5-,6-,9-/m1/s1/f/h14,19-20,22,24H,11H2 plec-1 Autophosphorylation of PleC 742036 N/A 742042 putative self putative self putative self putative self region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) phosphotransfer phosphotransfer reaction phosphotransfer false true false Fig. 2A Phosphorylation of PleC, Supp. Fig. 1C plec-divk-1 Autophosphorylation of pleC stimulated by divK 742036 N/A 742042 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742043 stimulator stimulator neutral component neutral component Q45976:p.Asp53Asn mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false Q45976:p.[Asp53Asn;Asp90Gly] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) phosphotransfer phosphotransfer reaction phosphotransfer false false false Fig. 2A, 2C, Supp. Fig 1A & B DivKD53N stimulated levels of PleC autophosphorylation in a concentration dependent manner plec-pled-1 Phosphorylation of PleD by PleC 742036 N/A 742042 enzyme enzyme neutral component neutral component region (required) phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742044 enzyme target enzyme target substrate substrate neutral component neutral component ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false Fig. 2B plec-pled-2 Phosphorylation of PleD by PleC stimulated by Divk 742036 N/A 742042 phosphate donor phosphate donor neutral component neutral component region (required) phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742043 stimulator stimulator ancillary ancillary Q45976:p.Asp53Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742044 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) phosphotransfer phosphotransfer reaction phosphotransfer false false false Fig. 2B plec-divk-2 Phosphorylation of DivK by PleC stimulated by Divk 742036 N/A 742043 enzyme target enzyme target substrate substrate neutral component neutral component ecoli Escherichia coli (strain K12) N/A 742043 stimulator stimulator ancillary ancillary Q45976:p.Asp53Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742042 enzyme enzyme neutral component neutral component region (required) phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false Fig. 3A, Supp. Fig 5 plec-divk-3 Phosphorylation of DivK by PleC 742036 N/A 742042 enzyme enzyme neutral component neutral component region (required) phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742043 enzyme target enzyme target substrate substrate neutral component neutral component ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false Fig. 3A divj-divk-1 Phosphorylation of DivK by DivJ 742037 N/A 742045 enzyme enzyme neutral component neutral component region (required) phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742043 enzyme target enzyme target substrate substrate neutral component neutral component ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false Fig. 2D, 4B divj-pled-1 Phosphorylation of PleD by DivJ 742037 N/A 742044 enzyme target enzyme target substrate substrate neutral component neutral component ecoli Escherichia coli (strain K12) N/A 742045 enzyme enzyme neutral component neutral component region (required) phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false Fig. 2D, 4B divj-divk-4 Autophosphorylation of DivJ stimulated by DivK 742037 N/A 742043 stimulator stimulator ancillary ancillary Q45976:p.Asp53Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false Q45976:p.[Asp53Asn;Asp90Gly] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 742045 putative self putative self putative self putative self region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false Fig. 4A, Supp. Fig 1C A asp53asn, asp90gly mutant of DivK was also included in the assay but the author did not comment on any effect. divj-1 Autophosphorylation of DivJ 742037 N/A 742045 putative self putative self putative self putative self region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false true false Fig. 4A, Supp. Fig. 1C divj-divk-3 Phosphorylation of DivK by DivJ stimulated by DivK 742037 N/A 742045 enzyme enzyme neutral component neutral component region (required) phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742043 stimulator stimulator ancillary ancillary Q45976:p.Asp53Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742043 enzyme target enzyme target substrate substrate neutral component neutral component ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false Fig. 4B divj-pled-2 Phosphorylation of PleD by DivJ stimulated by DivK 742037 N/A 742043 unspecified role unspecified role ancillary ancillary Q45976:p.Asp53Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742045 enzyme enzyme neutral component neutral component region (required) phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742044 enzyme target enzyme target substrate substrate neutral component neutral component ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false Fig. 4B plec-divk-4 Dephosphorylation of DivK by PleC 742038 N/A 742042 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742043 enzyme target enzyme target substrate substrate neutral component neutral component region phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) dephosphorylation dephosphorylation reaction dephosphorylation false false false Fig. 3B plec-divk-5 Dephosphorylation of DivK by PleC, inhibition by DivK 742038 N/A 742043 enzyme target enzyme target substrate substrate neutral component neutral component region phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 742042 enzyme enzyme neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742043 inhibitor inhibitor ancillary ancillary Q45976:p.Asp53Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) dephosphorylation dephosphorylation reaction dephosphorylation false false false Fig. 3B, 3C pled-1 DImerization of PleD 742039 N/A 742044 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false pled-plec-2 DImerization of PleD stimulated by PleC and DivK 742039 N/A 742044 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 742042 stimulator stimulator ancillary ancillary region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742043 stimulator stimulator ancillary ancillary Q45976:p.Asp53Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false Whilst there was low level of dimerization of PleD in the ansence of PleC and DivK, the presence of these proteins stimulated dimer formation plec-2 Dimerization of PleC 742040 N/A 742042 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Supp. Fig. 3 The distribution of PleC monomer and dimer fraction was not affected by preincubation with DivK. divj-2 Dimerization of Divj 742040 N/A 742045 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Supp. Fig. 4 The DivJ dimer fraction was not affected by preincubation with DivK. pled-plec-1 Stimulation of Caulobacter crescentus PleD activity by PleC and DivK 742041 N/A 742046 enzyme target enzyme target substrate substrate neutral component neutral component N/A 742042 stimulator stimulator ancillary ancillary P37894:p.Phe778Leu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742044 enzyme enzyme neutral component neutral component Q9A5I5:p.Asp53Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 742043 unspecified role unspecified role ancillary ancillary Q45976:p.Asp53Asn mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) enzymatic reaction enzymatic reaction Biochemical reaction Biochemical reaction false false false Fig. 1 The single domain response regulator DivK is able to efficiently stimulate the in vitro activity of PleD and that this activation requires an active histidine protein kinase PleC.