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/ hobbs-2012-3 Conserved small protein associates with the multidrug efflux pump AcrB and differentially affects antibiotic resistance. ecoli Escherichia coli (strain K12) adenylate cyclase adenylate cyclase complementation bacterial two-hybrid bacterial two-hybrid adenylate cyclase full dna sequence full identification by DNA sequencing full dna sequence Hobbs E.C., Yin X., Paul B.J., Astarita J.L., Storz G. storzg@mail.nih.gov Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2012 Only protein-protein interactions imex curation hobbs-2012-1 Conserved small protein associates with the multidrug efflux pump AcrB and differentially affects antibiotic resistance. ecoli Escherichia coli (strain K12) pull down pull down ms participant Identification by mass spectrometry ms participant Hobbs E.C., Yin X., Paul B.J., Astarita J.L., Storz G. storzg@mail.nih.gov Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2012 Only protein-protein interactions imex curation hobbs-2012-2 Conserved small protein associates with the multidrug efflux pump AcrB and differentially affects antibiotic resistance. ecoli Escherichia coli (strain K12) pull down pull down western blot western blot Immuno blot Immuno blot Hobbs E.C., Yin X., Paul B.J., Astarita J.L., Storz G. storzg@mail.nih.gov Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2012 Only protein-protein interactions imex curation acrb_ecoli Multidrug efflux pump subunit AcrB acrB acrE b0462 JW0451 AcrAB-TolC multidrug efflux pump subunit AcrB Acridine resistance protein B protein protein ecoli Escherichia coli (strain K12) MPNFFIDRPIFAWVIAIIIMLAGGLAILKLPVAQYPTIAPPAVTISASYPGADAKTVQDTVTQVIEQNMNGIDNLMYMSSNSDSTGTVQITLTFESGTDADIAQVQVQNKLQLAMPLLPQEVQQQGVSVEKSSSSFLMVVGVINTDGTMTQEDISDYVAANMKDAISRTSGVGDVQLFGSQYAMRIWMNPNELNKFQLTPVDVITAIKAQNAQVAAGQLGGTPPVKGQQLNASIIAQTRLTSTEEFGKILLKVNQDGSRVLLRDVAKIELGGENYDIIAEFNGQPASGLGIKLATGANALDTAAAIRAELAKMEPFFPSGLKIVYPYDTTPFVKISIHEVVKTLVEAIILVFLVMYLFLQNFRATLIPTIAVPVVLLGTFAVLAAFGFSINTLTMFGMVLAIGLLVDDAIVVVENVERVMAEEGLPPKEATRKSMGQIQGALVGIAMVLSAVFVPMAFFGGSTGAIYRQFSITIVSAMALSVLVALILTPALCATMLKPIAKGDHGEGKKGFFGWFNRMFEKSTHHYTDSVGGILRSTGRYLVLYLIIVVGMAYLFVRLPSSFLPDEDQGVFMTMVQLPAGATQERTQKVLNEVTHYYLTKEKNNVESVFAVNGFGFAGRGQNTGIAFVSLKDWADRPGEENKVEAITMRATRAFSQIKDAMVFAFNLPAIVELGTATGFDFELIDQAGLGHEKLTQARNQLLAEAAKHPDMLTSVRPNGLEDTPQFKIDIDQEKAQALGVSINDINTTLGAAWGGSYVNDFIDRGRVKKVYVMSEAKYRMLPDDIGDWYVRAADGQMVPFSAFSSSRWEYGSPRLERYNGLPSMEILGQAAPGKSTGEAMELMEQLASKLPTGVGYDWTGMSYQERLSGNQAPSLYAISLIVVFLCLAALYESWSIPFSVMLVVPLGVIGALLAATFRGLTNDVYFQVGLLTTIGLSAKNAILIVEFAKDLMDKEGKGLIEATLDAVRMRLRPILMTSLAFILGVMPLVISTGAGSGAQNAVGTGVMGGMVTATVLAIFFVPVFFVVVRRRFSRKNEDIEHSHTVDHH acrz_ecoli Multidrug efflux pump accessory protein AcrZ acrZ AcrAB-TolC multidrug efflux pump accessory protein AcrZ Acridine resistance protein Z ybhT JW5102 b0762 protein protein ecoli Escherichia coli (strain K12) MLELLKSLVFAVIMVPVVMAIILGLIYGLGEVFNIFSGVGKKDQPGQNH acra_ecoli Multidrug efflux pump subunit AcrA acrA lir mtcA b0463 JW0452 AcrAB-TolC multidrug efflux pump subunit AcrA Acridine resistance protein A protein protein ecoli Escherichia coli (strain K12) MNKNRGFTPLAVVLMLSGSLALTGCDDKQAQQGGQQMPAVGVVTVKTEPLQITTELPGRTSAYRIAEVRPQVSGIILKRNFKEGSDIEAGVSLYQIDPATYQATYDSAKGDLAKAQAAANIAQLTVNRYQKLLGTQYISKQEYDQALADAQQANAAVTAAKAAVETARINLAYTKVTSPISGRIGKSNVTEGALVQNGQATALATVQQLDPIYVDVTQSSNDFLRLKQELANGTLKQENGKAKVSLITSDGIKFPQDGTLEFSDVTVDQTTGSITLRAIFPNPDHTLLPGMFVRARLEEGLNPNAILVPQQGVTRTPRGDATVLVVGADDKVETRPIVASQAIGDKWLVTEGLKAGDRVVISGLQKVRPGVQVKAQEVTADNNQQAASGAQPEQSKS 5B81DD5BC2B0A077 acrb-ybht-5 1321609 n/a 1321612 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation t 25 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1321613 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation P0AAW9:p.Gly30Arg mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false t 18 fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Figure 4 acrb-ybht-6 1321609 n/a 1321613 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation P0AAW9:p.Gly30Arg mutation mutation certain certain sequence position certain certain certain sequence position certain false t 18 fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1321612 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation t 25 fusion protein fusion protein n-term range n-terminal range n-term range n-term range n-terminal range n-term range false P31224:p.His526Tyr mutation mutation certain certain sequence position certain certain certain sequence position certain false P31224:p.Leu984Pro mutation mutation certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 4A β-galactosidase activity in strains coexpressing AcrZG30R–T18 with T25–AcrBH526Y or AcrZG30R–T18 with T25–AcrBL984P was increased to 23% (for H526Y) or 37% (for L984P) of wild-type levels ybht-acra 1321610 n/a 1321612 unspecified role unspecified role prey prey n/a 1321613 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation spa tag tandem tag tandem tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1321614 unspecified role unspecified role prey prey association association false false false Figure 2 acrb-ybht-1 1321611 n/a 1321613 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation spa tag tandem tag tandem tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false P0AAW9:p.Gly30Arg mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false n/a 1321612 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation histag 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 false physical association physical association false false false Figure 3A AcrZ–SPA was retained by AcrB–His6 on Ni2+-NTA resin in the absence of both AcrA and TolC