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/ glover-2001-3 tau binds and organizes Escherichia coli replication proteins through distinct domains: domain III, shared by gamma and tau, oligomerizes DnaX. in vitro In vitro pull down pull down predetermined predetermined participant predetermined Glover B.P., Pritchard A.E., McHenry C.S. Crosslinking was followed by molecular sieving. Participant visualisation by biotin-tag binding to streptavidin-HRP and ECL detection (for domain detection) and anti-gamma Western Blot (for gamma and tau) charles.mchenry@uchsc.edu J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation glover-2001-1 tau binds and organizes Escherichia coli replication proteins through distinct domains: domain III, shared by gamma and tau, oligomerizes DnaX. in vitro In vitro crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Glover B.P., Pritchard A.E., McHenry C.S. Crosslinking was followed by molecular sieving. Participant visualisation by biotin-tag binding to streptavidin-HRP and ECL detection (for domain detection) and anti-gamma Western Blot (for gamma and tau) charles.mchenry@uchsc.edu J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation glover-2001-2 tau binds and organizes Escherichia coli replication proteins through distinct domains: domain III, shared by gamma and tau, oligomerizes DnaX. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Glover B.P., Pritchard A.E., McHenry C.S. Crosslinking was followed by molecular sieving. Participant visualisation by biotin-tag binding to streptavidin-HRP and ECL detection (for domain detection) and anti-gamma Western Blot (for gamma and tau) charles.mchenry@uchsc.edu J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation glover-2001-4 tau binds and organizes Escherichia coli replication proteins through distinct domains: domain III, shared by gamma and tau, oligomerizes DnaX. in vitro In vitro crosslink cross-linking study crosslink anti tag western anti tag western blot anti tag western Glover B.P., Pritchard A.E., McHenry C.S. Crosslinking was followed by molecular sieving. Participant visualisation by biotin-tag binding to streptavidin-HRP and ECL detection (for domain detection) and anti-gamma Western Blot (for gamma and tau) charles.mchenry@uchsc.edu J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation holb_ecoli DNA polymerase III subunit delta' holB b1099 JW1085 protein protein ecoli Escherichia coli (strain K12) MRWYPWLRPDFEKLVASYQAGRGHHALLIQALPGMGDDALIYALSRYLLCQQPQGHKSCGHCRGCQLMQAGTHPDYYTLAPEKGKNTLGVDAVREVTEKLNEHARLGGAKVVWVTDAALLTDAAANALLKTLEEPPAETWFFLATREPERLLATLRSRCRLHYLAPPPEQYAVTWLSREVTMSQDALLAALRLSAGSPGAALALFQGDNWQARETLCQALAYSVPSGDWYSLLAALNHEQAPARLHWLATLLMDALKRHHGAAQVTNVDVPGLVAELANHLSPSRLQAILGDVCHIREQLMSVTGINRELLITDLLLRIEHYLQPGVVLPVPHL dpo3x_ecoli DNA polymerase III subunit tau dnaX dnaZ dnaZX b0470 JW0459 DNA polymerase III subunit gamma protein protein ecoli Escherichia coli (strain K12) MSYQVLARKWRPQTFADVVGQEHVLTALANGLSLGRIHHAYLFSGTRGVGKTSIARLLAKGLNCETGITATPCGVCDNCREIEQGRFVDLIEIDAASRTKVEDTRDLLDNVQYAPARGRFKVYLIDEVHMLSRHSFNALLKTLEEPPEHVKFLLATTDPQKLPVTILSRCLQFHLKALDVEQIRHQLEHILNEEHIAHEPRALQLLARAAEGSLRDALSLTDQAIASGDGQVSTQAVSAMLGTLDDDQALSLVEAMVEANGERVMALINEAAARGIEWEALLVEMLGLLHRIAMVQLSPAALGNDMAAIELRMRELARTIPPTDIQLYYQTLLIGRKELPYAPDRRMGVEMTLLRALAFHPRMPLPEPEVPRQSFAPVAPTAVMTPTQVPPQPQSAPQQAPTVPLPETTSQVLAARQQLQRVQGATKAKKSEPAAATRARPVNNAALERLASVTDRVQARPVPSALEKAPAKKEAYRWKATTPVMQQKEVVATPKALKKALEHEKTPELAAKLAAEAIERDPWAAQVSQLSLPKLVEQVALNAWKEESDNAVCLHLRSSQRHLNNRGAQQKLAEALSMLKGSTVELTIVEDDNPAVRTPLEWRQAIYEEKLAQARESIIADNNIQTLRRFFDAELDEESIRPI holc_ecoli DNA polymerase III subunit chi holC b4259 JW4216 protein protein ecoli Escherichia coli (strain K12) MKNATFYLLDNDTTVDGLSAVEQLVCEIAAERWRSGKRVLIACEDEKQAYRLDEALWARPAESFVPHNLAGEGPRGGAPVEIAWPQKRSSSRRDILISLRTSFADFATAFTEVVDFVPYEDSLKQLARERYKAYRVAGFNLNTATWK 7FEFFC359B28AA88 hold_ecoli DNA polymerase III subunit psi holD b4372 JW4334 protein protein ecoli Escherichia coli (strain K12) MTSRRDWQLQQLGITQWSLRRPGALQGEIAIAIPAHVRLVMVANDLPALTDPLVSDVLRALTVSPDQVLQLTPEKIAMLPQGSHCNSWRLGTDEPLSLEGAQVASPALTDLRANPTARAALWQQICTYEHDFFPRND hola_ecoli DNA polymerase III subunit delta b0640 holA JW0635 protein protein ecoli Escherichia coli (strain K12) MIRLYPEQLRAQLNEGLRAAYLLLGNDPLLLQESQDAVRQVAAAQGFEEHHTFSIDPNTDWNAIFSLCQAMSLFASRQTLLLLLPENGPNAAINEQLLTLTGLLHDDLLLIVRGNKLSKAQENAAWFTALANRSVQVTCQTPEQAQLPRWVAARAKQLNLELDDAANQVLCYCYEGNLLALAQALERLSLLWPDGKLTLPRVEQAVNDAAHFTPFHWVDALLMGKSKRALHILQQLRLEGSEPVILLRTLQRELLLLVNLKRQSAHTPLRALFDKHRVWQNRRGMMGEALNRLSQTQLRQAVQLLTRTELTLKQDYGQSVWAELEGLSLLLCHKPLADVFIDG 87239A678FE16BE2 p06710-1 DNA polymerase III subunit tau dnaX dnaZ dnaZX DNA polymerase III subunit gamma b0470 JW0459 protein protein ecoli Escherichia coli (strain K12) MSYQVLARKWRPQTFADVVGQEHVLTALANGLSLGRIHHAYLFSGTRGVGKTSIARLLAKGLNCETGITATPCGVCDNCREIEQGRFVDLIEIDAASRTKVEDTRDLLDNVQYAPARGRFKVYLIDEVHMLSRHSFNALLKTLEEPPEHVKFLLATTDPQKLPVTILSRCLQFHLKALDVEQIRHQLEHILNEEHIAHEPRALQLLARAAEGSLRDALSLTDQAIASGDGQVSTQAVSAMLGTLDDDQALSLVEAMVEANGERVMALINEAAARGIEWEALLVEMLGLLHRIAMVQLSPAALGNDMAAIELRMRELARTIPPTDIQLYYQTLLIGRKELPYAPDRRMGVEMTLLRALAFHPRMPLPEPEVPRQSFAPVAPTAVMTPTQVPPQPQSAPQQAPTVPLPETTSQVLAARQQLQRVQGATKAKKSEPAAATRARPVNNAALERLASVTDRVQARPVPSALEKAPAKKEAYRWKATTPVMQQKEVVATPKALKKALEHEKTPELAAKLAAEAIERDPWAAQVSQLSLPKLVEQVALNAWKEESDNAVCLHLRSSQRHLNNRGAQQKLAEALSMLKGSTVELTIVEDDNPAVRTPLEWRQAIYEEKLAQARESIIADNNIQTLRRFFDAELDEESIRPI Produced by full-length translation of the dnaX gene. This isoform is essential, constructs that express only the gamma isoform are not viable (Ref.14). p06710-2 DNA polymerase III subunit tau dnaX dnaZ dnaZX b0470 JW0459 DNA polymerase III subunit gamma protein protein ecoli Escherichia coli (strain K12) MSYQVLARKWRPQTFADVVGQEHVLTALANGLSLGRIHHAYLFSGTRGVGKTSIARLLAKGLNCETGITATPCGVCDNCREIEQGRFVDLIEIDAASRTKVEDTRDLLDNVQYAPARGRFKVYLIDEVHMLSRHSFNALLKTLEEPPEHVKFLLATTDPQKLPVTILSRCLQFHLKALDVEQIRHQLEHILNEEHIAHEPRALQLLARAAEGSLRDALSLTDQAIASGDGQVSTQAVSAMLGTLDDDQALSLVEAMVEANGERVMALINEAAARGIEWEALLVEMLGLLHRIAMVQLSPAALGNDMAAIELRMRELARTIPPTDIQLYYQTLLIGRKELPYAPDRRMGVEMTLLRALAFHPRMPLPEPEVPRQSFAPVAPTAVMTPTQVPPQPQSAPQQAPTVPLPETTSQVLAARQQLQRVQGATKAKKE Formed by programmed ribosomal frameshifting to a premature stop codon in the -1 frame at codon 430, the last residue is thus Glu and not Ser. Mutants which remove the frameshift are viable, suggesting strongly that gamma is not essential for viability (Ref.14). tau complex 1349886 n/a 1349890 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) n/a 1349891 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 false n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false 222-643 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1349892 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) n/a 1349893 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) n/a 1349891 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) n/a 1349894 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) association association false false false Fig. 4a tau complex, part of DNA Polymerase complex domain iii-tau-chi-psi 1349886 n/a 1349891 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 false 222-643 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) n/a 1349893 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) n/a 1349892 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) n/a 1349891 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) association association false false false Fig. 4a domain iii-chi-psi 1349886 n/a 1349893 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) n/a 1349891 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 false 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 false 222-643 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1349892 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) association association false false false Fig. 4a tau dimer 1349887 n/a 1349895 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig. 2 dimer gamma dimer 1349887 n/a 1349896 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig. 2 dimer domain iii-v 1349888 n/a 1349891 unspecified role unspecified role neutral component neutral component n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false 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 false 222-643 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1349891 unspecified role unspecified role neutral component neutral component 222-643 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false 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 false n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) 1349932 1349930 direct interaction direct interaction false false false Fig. 3a, b, e multimer formation according to Stoke radius measure domain iii-v 1349889 n/a 1349891 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 false 222-643 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig. 2 dimer