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/ A three-domain structure for the delta subunit of the DNA polymerase III holoenzyme delta domain III binds delta' and assembles into the DnaX complex. A three-domain structure for the delta subunit of the DNA polymerase III holoenzyme delta domain III binds delta' and assembles into the DnaX complex. J. Biol. Chem. (0021-9258) 2002 imex curation Bullard J.M., Pritchard A.E., Song M.S., Glover B.P., Wieczorek A., Chen J., Janjic N., McHenry C.S. charles.mchenry@ushsc.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 Bullard J.M., Pritchard A.E., Song M.S., Glover B.P., Wieczorek A., Chen J., Janjic N., McHenry C.S. charles.mchenry@ushsc.edu Accepted 2013-JAN-04 by ORCHARD 04-Feb-2013: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2002 Only protein-protein interactions imex curation A three-domain structure for the delta subunit of the DNA polymerase III holoenzyme delta domain III binds delta' and assembles into the DnaX complex. A three-domain structure for the delta subunit of the DNA polymerase III holoenzyme delta domain III binds delta' and assembles into the DnaX complex. J. Biol. Chem. (0021-9258) 2002 imex curation Bullard J.M., Pritchard A.E., Song M.S., Glover B.P., Wieczorek A., Chen J., Janjic N., McHenry C.S. charles.mchenry@ushsc.edu 04-Feb-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Bullard J.M., Pritchard A.E., Song M.S., Glover B.P., Wieczorek A., Chen J., Janjic N., McHenry C.S. charles.mchenry@ushsc.edu Accepted 2013-JAN-04 by ORCHARD 04-Feb-2013: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2002 Only protein-protein interactions imex curation A three-domain structure for the delta subunit of the DNA polymerase III holoenzyme delta domain III binds delta' and assembles into the DnaX complex. A three-domain structure for the delta subunit of the DNA polymerase III holoenzyme delta domain III binds delta' and assembles into the DnaX complex. J. Biol. Chem. (0021-9258) 2002 imex curation Bullard J.M., Pritchard A.E., Song M.S., Glover B.P., Wieczorek A., Chen J., Janjic N., McHenry C.S. charles.mchenry@ushsc.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 Bullard J.M., Pritchard A.E., Song M.S., Glover B.P., Wieczorek A., Chen J., Janjic N., McHenry C.S. charles.mchenry@ushsc.edu Accepted 2013-JAN-04 by ORCHARD 04-Feb-2013: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2002 Only protein-protein interactions imex curation 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 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 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 hola domain iii-holb 2 7 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 5 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 207-343 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 5, Table II KD value is for interaction of holA domain III (P28630) with full-length holB (P28631) hola-gamma 1 5 unspecified role unspecified role neutral component neutral component c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminal biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association Fig. 7 hola-tau 1 4 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component c-terminal biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) physical association physical association Fig. 7 hola domain iii-holb-gamma 3 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 5 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 207-343 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association Fig. 6 KD value is for interaction of holA domain III (P28630) with full-length holB (P28631)