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/ Mutational analysis of beta '260-309, a sigma 70 binding site located on Escherichia coli core RNA polymerase. Mutational analysis of beta '260-309, a sigma 70 binding site located on Escherichia coli core RNA polymerase. J. Biol. Chem. (0021-9258) 2000 imex curation Arthur TM., Anthony LC., Burgess RR. burgess@oncology.wisc.edu 16-Dec-2008: Contacted by JYOTI. davet Only protein-protein interactions ecoli Escherichia coli (strain K12) pull down pull down western blot western blot Immuno blot Immuno blot Arthur TM., Anthony LC., Burgess RR. burgess@oncology.wisc.edu Accepted 2008-Not a month!-2 by DAVET. 16-Dec-2008: Contacted by JYOTI. phase of growth J. Biol. Chem. (0021-9258) 2000 davet Only protein-protein interactions imex curation Mutational analysis of beta '260-309, a sigma 70 binding site located on Escherichia coli core RNA polymerase. Mutational analysis of beta '260-309, a sigma 70 binding site located on Escherichia coli core RNA polymerase. J. Biol. Chem. (0021-9258) 2000 imex curation Arthur TM., Anthony LC., Burgess RR. burgess@oncology.wisc.edu 16-Dec-2008: Contacted by JYOTI. davet Only protein-protein interactions in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting western blot western blot Immuno blot Immuno blot Arthur TM., Anthony LC., Burgess RR. burgess@oncology.wisc.edu Accepted 2008-Not a month!-2 by DAVET. 16-Dec-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2000 davet Only protein-protein interactions imex curation rpod_ecoli RNA polymerase sigma factor RpoD rpoD alt b3067 Sigma-70 JW3039 protein protein ecoli Escherichia coli (strain K12) MEQNPQSQLKLLVTRGKEQGYLTYAEVNDHLPEDIVDSDQIEDIIQMINDMGIQVMEEAPDADDLMLAENTADEDAAEAAAQVLSSVESEIGRTTDPVRMYMREMGTVELLTREGEIDIAKRIEDGINQVQCSVAEYPEAITYLLEQYDRVEAEEARLSDLITGFVDPNAEEDLAPTATHVGSELSQEDLDDDEDEDEEDGDDDSADDDNSIDPELAREKFAELRAQYVVTRDTIKAKGRSHATAQEEILKLSEVFKQFRLVPKQFDYLVNSMRVMMDRVRTQERLIMKLCVEQCKMPKKNFITLFTGNETSDTWFNAAIAMNKPWSEKLHDVSEEVHRALQKLQQIEEETGLTIEQVKDINRRMSIGEAKARRAKKEMVEANLRLVISIAKKYTNRGLQFLDLIQEGNIGLMKAVDKFEYRRGYKFSTYATWWIRQAITRSIADQARTIRIPVHMIETINKLNRISRQMLQEMGREPTPEELAERMLMPEDKIRKVLKIAKEPISMETPIGDDEDSHLGDFIEDTTLELPLDSATTESLRAATHDVLAGLTAREAKVLRMRFGIDMNTDYTLEEVGKQFDVTRERIRQIEAKALRKLRHPSRSEVLRSFLDD DIP protein P00579 original sequence version: 148 DIP protein P00579 original sequence version: 114 DIP protein P00579 original sequence version: 162 DIP protein P00579 original sequence version: 136 DIP protein P00579 original sequence version: null CA4F0E30DEC1703D rpoa_ecoli DNA-directed RNA polymerase subunit alpha rpoA pez phs sez b3295 Transcriptase subunit alpha RNA polymerase subunit alpha JW3257 protein protein ecoli Escherichia coli (strain K12) MQGSVTEFLKPRLVDIEQVSSTHAKVTLEPLERGFGHTLGNALRRILLSSMPGCAVTEVEIDGVLHEYSTKEGVQEDILEILLNLKGLAVRVQGKDEVILTLNKSGIGPVTAADITHDGDVEIVKPQHVICHLTDENASISMRIKVQRGRGYVPASTRIHSEEDERPIGRLLVDACYSPVERIAYNVEAARVEQRTDLDKLVIEMETNGTIDPEEAIRRAATILAEQLEAFVDLRDVRQPEVKEEKPEFDPILLRPVDDLELTVRSANCLKAEAIHYIGDLVQRTEVELLKTPNLGKKSLTEIKDVLASRGLSLGMRLENWPPASIADE DIP protein P0A7Z4 original sequence version: 55 DIP protein P0A7Z4 original sequence version: 60 DIP protein P0A7Z4 original sequence version: 69 DIP protein P0A7Z4 original sequence version: 107 DIP protein P0A7Z4 original sequence version: 93 12A14B75A3CAEA19 rpoc_ecoli DNA-directed RNA polymerase subunit beta' rpoC tabB b3988 JW3951 Transcriptase subunit beta' RNA polymerase subunit beta' protein protein ecoli Escherichia coli (strain K12) MKDLLKFLKAQTKTEEFDAIKIALASPDMIRSWSFGEVKKPETINYRTFKPERDGLFCARIFGPVKDYECLCGKYKRLKHRGVICEKCGVEVTQTKVRRERMGHIELASPTAHIWFLKSLPSRIGLLLDMPLRDIERVLYFESYVVIEGGMTNLERQQILTEEQYLDALEEFGDEFDAKMGAEAIQALLKSMDLEQECEQLREELNETNSETKRKKLTKRIKLLEAFVQSGNKPEWMILTVLPVLPPDLRPLVPLDGGRFATSDLNDLYRRVINRNNRLKRLLDLAAPDIIVRNEKRMLQEAVDALLDNGRRGRAITGSNKRPLKSLADMIKGKQGRFRQNLLGKRVDYSGRSVITVGPYLRLHQCGLPKKMALELFKPFIYGKLELRGLATTIKAAKKMVEREEAVVWDILDEVIREHPVLLNRAPTLHRLGIQAFEPVLIEGKAIQLHPLVCAAYNADFDGDQMAVHVPLTLEAQLEARALMMSTNNILSPANGEPIIVPSQDVVLGLYYMTRDCVNAKGEGMVLTGPKEAERLYRSGLASLHARVKVRITEYEKDANGELVAKTSLKDTTVGRAILWMIVPKGLPYSIVNQALGKKAISKMLNTCYRILGLKPTVIFADQIMYTGFAYAARSGASVGIDDMVIPEKKHEIISEAEAEVAEIQEQFQSGLVTAGERYNKVIDIWAAANDRVSKAMMDNLQTETVINRDGQEEKQVSFNSIYMMADSGARGSAAQIRQLAGMRGLMAKPDGSIIETPITANFREGLNVLQYFISTHGARKGLADTALKTANSGYLTRRLVDVAQDLVVTEDDCGTHEGIMMTPVIEGGDVKEPLRDRVLGRVTAEDVLKPGTADILVPRNTLLHEQWCDLLEENSVDAVKVRSVVSCDTDFGVCAHCYGRDLARGHIINKGEAIGVIAAQSIGEPGTQLTMRTFHIGGAASRAAAESSIQVKNKGSIKLSNVKSVVNSSGKLVITSRNTELKLIDEFGRTKESYKVPYGAVLAKGDGEQVAGGETVANWDPHTMPVITEVSGFVRFTDMIDGQTITRQTDELTGLSSLVVLDSAERTAGGKDLRPALKIVDAQGNDVLIPGTDMPAQYFLPGKAIVQLEDGVQISSGDTLARIPQESGGTKDITGGLPRVADLFEARRPKEPAILAEISGIVSFGKETKGKRRLVITPVDGSDPYEEMIPKWRQLNVFEGERVERGDVISDGPEAPHDILRLRGVHAVTRYIVNEVQDVYRLQGVKINDKHIEVIVRQMLRKATIVNAGSSDFLEGEQVEYSRVKIANRELEANGKVGATYSRDLLGITKASLATESFISAASFQETTRVLTEAAVAGKRDELRGLKENVIVGRLIPAGTGYAYHQDRMRRRAAGEAPAAPQVTAEDASASLAELLNAGLGGSDNE DIP protein P0A8T7 original sequence version: 50 DIP protein P0A8T7 original sequence version: 55 DIP protein P0A8T7 original sequence version: 105 DIP protein P0A8T7 original sequence version: 89 A6878C61D15F4961 rpoc-rpod 2 5 unspecified role unspecified role bait bait P0A8T7:p.Arg293Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R Q Wed May 02 14:28:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg293gln' Tue Jun 30 16:01:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg293Gln' n terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P0A8T7:p.Arg275Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R Q Wed May 02 18:54:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg275gln' Tue Jun 30 19:03:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg275Gln' P0A8T7:p.Glu295Lys mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain E K Wed May 02 19:44:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu295lys' Tue Jun 30 17:08:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu295Lys' P0A8T7:p.Arg297Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R S Wed May 02 15:38:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg297ser' Tue Jun 30 19:00:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg297Ser' P0A8T7:p.Ala302Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain A D Wed May 02 14:33:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala302asp' Tue Jun 30 19:19:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala302Asp' P0A8T7:p.Gln300Glu mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain Q E Wed May 02 19:07:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln300glu' Tue Jun 30 17:41:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln300Glu' P0A8T7:p.Asn309Asp mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain N D Wed May 02 19:57:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn309asp' Tue Jun 30 18:33:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn309Asp' ecoli Escherichia coli (strain K12) 3 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) physical association physical association 2 Five of the mutations (R275Q, R293Q, E295K, R297S, and A302D) were greatly reduced in their ability to bind sigma 70. rpoc-rpod-1 1 5 unspecified role unspecified role bait bait P0A8T7:p.Arg293Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R Q Wed May 02 19:28:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg293gln' Tue Jun 30 15:42:22 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg293Gln' P0A8T7:p.Arg275Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R Q Wed May 02 15:24:41 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg275gln' Tue Jun 30 20:03:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg275Gln' P0A8T7:p.Glu295Lys mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain E K Wed May 02 15:01:21 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu295lys' Tue Jun 30 19:08:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu295Lys' P0A8T7:p.Arg297Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R S Wed May 02 19:25:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg297ser' Tue Jun 30 17:26:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg297Ser' P0A8T7:p.Ala302Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain A D Wed May 02 20:05:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala302asp' Tue Jun 30 19:51:56 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala302Asp' region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P0A8T7:p.Gln300Glu mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain Q E Wed May 02 20:25:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln300glu' Tue Jun 30 19:46:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln300Glu' 4 unspecified role unspecified role prey prey 3 unspecified role unspecified role prey prey physical association physical association 4 mutations R275Q, E295K, and A302D caused defects in binding sigma 70 in both log and stationary phase samples (Fig. 4C). Also reduced in Esigma 70 formation were N266D in both log and stationary phase samples and R297S in log phase samples. Q300E again showed properties of binding sigma 70 better than wild type. log phase rpoc-rpod-2 1 5 unspecified role unspecified role bait bait P0A8T7:p.Arg293Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R Q Wed May 02 18:52:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg293gln' Tue Jun 30 16:36:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg293Gln' P0A8T7:p.Arg275Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R Q Wed May 02 15:58:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg275gln' Tue Jun 30 18:52:38 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg275Gln' P0A8T7:p.Glu295Lys mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain E K Wed May 02 19:47:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu295lys' Tue Jun 30 16:49:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu295Lys' P0A8T7:p.Ala302Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain A D Wed May 02 14:31:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala302asp' Tue Jun 30 16:25:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala302Asp' region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P0A8T7:p.Gln300Glu mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain Q E Wed May 02 17:10:05 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln300glu' Tue Jun 30 15:55:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln300Glu' 3 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey physical association physical association 4 mutations R275Q, E295K, and A302D caused defects in binding sigma 70 in both log and stationary phase samples (Fig. 4C). Also reduced in Esigma 70 formation were N266D in both log and stationary phase samples and R297S in log phase samples. Q300E again showed properties of binding sigma 70 better than wild type. stationary phase