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/ tau binds and organizes Escherichia coli replication proteins through distinct domains: domain III, shared by gamma and tau, oligomerizes DnaX. tau binds and organizes Escherichia coli replication proteins through distinct domains: domain III, shared by gamma and tau, oligomerizes DnaX. J. Biol. Chem. (0021-9258) 2001 imex curation 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 04-Feb-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 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 Accepted 2013-JAN-04 by ORCHARD 04-Feb-2013: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation tau binds and organizes Escherichia coli replication proteins through distinct domains: domain III, shared by gamma and tau, oligomerizes DnaX. tau binds and organizes Escherichia coli replication proteins through distinct domains: domain III, shared by gamma and tau, oligomerizes DnaX. J. Biol. Chem. (0021-9258) 2001 imex curation 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 04-Feb-2013: Contacted by IntAct-Help. Only protein-protein interactions 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 Accepted 2013-JAN-04 by ORCHARD 04-Feb-2013: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation tau binds and organizes Escherichia coli replication proteins through distinct domains: domain III, shared by gamma and tau, oligomerizes DnaX. tau binds and organizes Escherichia coli replication proteins through distinct domains: domain III, shared by gamma and tau, oligomerizes DnaX. J. Biol. Chem. (0021-9258) 2001 imex curation 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 04-Feb-2013: Contacted by IntAct-Help. Only protein-protein interactions 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 Accepted 2013-JAN-04 by ORCHARD 04-Feb-2013: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation tau binds and organizes Escherichia coli replication proteins through distinct domains: domain III, shared by gamma and tau, oligomerizes DnaX. tau binds and organizes Escherichia coli replication proteins through distinct domains: domain III, shared by gamma and tau, oligomerizes DnaX. J. Biol. Chem. (0021-9258) 2001 imex curation 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 04-Feb-2013: Contacted by IntAct-Help. Only protein-protein interactions 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 Accepted 2013-JAN-04 by ORCHARD 04-Feb-2013: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation 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). 8177A07F31BC86E3 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). D2028BD99E375150 hola_ecoli DNA polymerase III subunit delta b0640 holA JW0635 protein protein ecoli Escherichia coli (strain K12) MIRLYPEQLRAQLNEGLRAAYLLLGNDPLLLQESQDAVRQVAAAQGFEEHHTFSIDPNTDWNAIFSLCQAMSLFASRQTLLLLLPENGPNAAINEQLLTLTGLLHDDLLLIVRGNKLSKAQENAAWFTALANRSVQVTCQTPEQAQLPRWVAARAKQLNLELDDAANQVLCYCYEGNLLALAQALERLSLLWPDGKLTLPRVEQAVNDAAHFTPFHWVDALLMGKSKRALHILQQLRLEGSEPVILLRTLQRELLLLVNLKRQSAHTPLRALFDKHRVWQNRRGMMGEALNRLSQTQLRQAVQLLTRTELTLKQDYGQSVWAELEGLSLLLCHKPLADVFIDG 87239A678FE16BE2 87239A678FE16BE2 hold_ecoli DNA polymerase III subunit psi holD b4372 JW4334 protein protein ecoli Escherichia coli (strain K12) MTSRRDWQLQQLGITQWSLRRPGALQGEIAIAIPAHVRLVMVANDLPALTDPLVSDVLRALTVSPDQVLQLTPEKIAMLPQGSHCNSWRLGTDEPLSLEGAQVASPALTDLRANPTARAALWQQICTYEHDFFPRND DIP protein P28632 original sequence version: 88 DIP protein P28632 original sequence version: null 5B7356201E7E27FE holb_ecoli DNA polymerase III subunit delta' holB b1099 JW1085 protein protein ecoli Escherichia coli (strain K12) MRWYPWLRPDFEKLVASYQAGRGHHALLIQALPGMGDDALIYALSRYLLCQQPQGHKSCGHCRGCQLMQAGTHPDYYTLAPEKGKNTLGVDAVREVTEKLNEHARLGGAKVVWVTDAALLTDAAANALLKTLEEPPAETWFFLATREPERLLATLRSRCRLHYLAPPPEQYAVTWLSREVTMSQDALLAALRLSAGSPGAALALFQGDNWQARETLCQALAYSVPSGDWYSLLAALNHEQAPARLHWLATLLMDALKRHHGAAQVTNVDVPGLVAELANHLSPSRLQAILGDVCHIREQLMSVTGINRELLITDLLLRIEHYLQPGVVLPVPHL DIP protein P28631 original sequence version: 95 DIP protein P28631 original sequence version: null DC9DAA644AD8A096 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 DIP protein P06710 original sequence version: 106 DIP protein P06710 original sequence version: null D2028BD99E375150 holc_ecoli DNA polymerase III subunit chi holC b4259 JW4216 protein protein ecoli Escherichia coli (strain K12) MKNATFYLLDNDTTVDGLSAVEQLVCEIAAERWRSGKRVLIACEDEKQAYRLDEALWARPAESFVPHNLAGEGPRGGAPVEIAWPQKRSSSRRDILISLRTSFADFATAFTEVVDFVPYEDSLKQLARERYKAYRVAGFNLNTATWK 7FEFFC359B28AA88 7FEFFC359B28AA88 domain iii-v 2 10 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 222-643 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 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. 2 dimer tau dimer 4 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 2 dimer gamma dimer 4 5 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 2 dimer domain iii-v 1 10 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 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 222-643 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 10 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 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 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) 27 25 direct interaction direct interaction Fig. 3a, b, e multimer formation according to Stoke radius measure tau complex 3 9 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 10 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 n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 222-643 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) association association Fig. 4a tau complex, part of DNA Polymerase complex domain iii-tau-chi-psi 3 10 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 222-643 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 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) 8 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) association association Fig. 4a domain iii-chi-psi 3 8 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 10 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 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 222-643 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) association association Fig. 4a