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/ Specific interactions with TBP and TFIIB in vitro suggest that 14-3-3 proteins may participate in the regulation of transcription when part of a DNA binding complex. Specific interactions with TBP and TFIIB in vitro suggest that 14-3-3 proteins may participate in the regulation of transcription when part of a DNA binding complex. Plant Cell (1040-4651) 1999 imex curation Pan S., Sehnke PC., Ferl RJ., Gurley WB. The VP16 protein mentioned in this study could not be unambiguously identified and therefore interactions involving this protein were not curated. wgurley@micro.ifas.ufl.edu 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Pan S., Sehnke PC., Ferl RJ., Gurley WB. The VP16 protein mentioned in this study could not be unambiguously identified and therefore interactions involving this protein were not curated. wgurley@micro.ifas.ufl.edu Accepted 2009-JUN-27 by JYOTI. Plant Cell (1040-4651) 1999 jyoti Only protein-protein interactions imex curation Specific interactions with TBP and TFIIB in vitro suggest that 14-3-3 proteins may participate in the regulation of transcription when part of a DNA binding complex. Specific interactions with TBP and TFIIB in vitro suggest that 14-3-3 proteins may participate in the regulation of transcription when part of a DNA binding complex. Plant Cell (1040-4651) 1999 imex curation Pan S., Sehnke PC., Ferl RJ., Gurley WB. The VP16 protein mentioned in this study could not be unambiguously identified and therefore interactions involving this protein were not curated. wgurley@micro.ifas.ufl.edu 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Pan S., Sehnke PC., Ferl RJ., Gurley WB. The VP16 protein mentioned in this study could not be unambiguously identified and therefore interactions involving this protein were not curated. wgurley@micro.ifas.ufl.edu Accepted 2009-JUN-27 by JYOTI. 21-Dec-2010: Contacted by JYOTI. Plant Cell (1040-4651) 1999 jyoti Only protein-protein interactions imex curation Specific interactions with TBP and TFIIB in vitro suggest that 14-3-3 proteins may participate in the regulation of transcription when part of a DNA binding complex. Specific interactions with TBP and TFIIB in vitro suggest that 14-3-3 proteins may participate in the regulation of transcription when part of a DNA binding complex. Plant Cell (1040-4651) 1999 imex curation Pan S., Sehnke PC., Ferl RJ., Gurley WB. The VP16 protein mentioned in this study could not be unambiguously identified and therefore interactions involving this protein were not curated. wgurley@micro.ifas.ufl.edu 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Pan S., Sehnke PC., Ferl RJ., Gurley WB. The VP16 protein mentioned in this study could not be unambiguously identified and therefore interactions involving this protein were not curated. wgurley@micro.ifas.ufl.edu Accepted 2009-JUN-27 by JYOTI. 21-Dec-2010: Contacted by JYOTI. Plant Cell (1040-4651) 1999 jyoti Only protein-protein interactions imex curation tf2b1_arath Transcription initiation factor IIB-1 TFIIB1 General transcription factor TFIIB-1 T32G6.15 At2g41630 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSDAYCTDCKKETELVVDHSAGDTLCSECGLVLESHSIDETSEWRTFANESSNSDPNRVGGPTNPLLADSALTTVIAKPNGSSGDFLSSSLGRWQNRNSNSDRGLIQAFKTIATMSERLGLVATIKDRANELYKRLEDQKSSRGRNQDALYAACLYIACRQEDKPRTIKEICVIANGATKKEIGRAKDYIVKTLGLEPGQSVDLGTIHAGDFMRRFCSNLAMSNHAVKAAQEAVQKSEEFDIRRSPISIAAVVIYIITQLSDDKKTLKDISHATGVAEGTIRNSYKDLYPHLSKIAPSWYAKEEDLKNLSSP D731D2FAE60484EA 14332_arath 14-3-3-like protein GF14 omega GRF2 GF14 At1g78300 F3F9.16 General regulatory factor 2 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MASGREEFVYMAKLAEQAERYEEMVEFMEKVSAAVDGDELTVEERNLLSVAYKNVIGARRASWRIISSIEQKEESRGNDDHVTAIREYRSKIETELSGICDGILKLLDSRLIPAAASGDSKVFYLKMKGDYHRYLAEFKTGQERKDAAEHTLAAYKSAQDIANAELAPTHPIRLGLALNFSVFYYEILNSPDRACNLAKQAFDEAIAELDTLGEESYKDSTLIMQLLRDNLTLWTSDMQDDAADEIKEAAAPKPTEEQQ 9CCE5043A707918A 14337_arath 14-3-3-like protein GF14 nu GRF7 At3g02520 F16B3.15 General regulatory factor 7 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSSSREENVYLAKLAEQAERYEEMVEFMEKVAKTVDTDELTVEERNLLSVAYKNVIGARRASWRIISSIEQKEESRGNDDHVSIIKDYRGKIETELSKICDGILNLLDSHLVPTASLAESKVFYLKMKGDYHRYLAEFKTGAERKEAAESTLVAYKSAQDIALADLAPTHPIRLGLALNFSVFYYEILNSPDRACSLAKQAFDEAISELDTLGEESYKDSTLIMQLLRDNLTLWNSDINDEAGGDEIKEASKHEPEEGKPAETGQ BF76ECB7F76E166A taf9_human Transcription initiation factor TFIID subunit 9 Transcription initiation factor TFIID 31 kDa subunit Transcription initiation factor TFIID 32 kDa subunit RNA polymerase II TBP-associated factor subunit G STAF31/32 TAF9 TAF2G TAFII31 protein protein human Homo sapiens Human MESGKTASPKSMPKDAQMMAQILKDMGITEYEPRVINQMLEFAFRYVTTILDDAKIYSSHAKKATVDADDVRLAIQCRADQSFTSPPPRDFLLDIARQRNQTPLPLIKPYSGPRLPPDRYCLTAPNYRLKSLQKKASTSAGRITVPRLSVGSVTSRPSTPTLGTPTPQTMSVSTKVGTPMSLTGQRFTVQMPTSQSPAVKASIPATSAVQNVLINPSLIGSKNILITTNMMSSQNTANESSNALKRKREDDDDDDDDDDDYDNL DIP protein Q16594 original sequence version: 66 DIP protein Q16594 original sequence version: 105 DIP protein Q16594 original sequence version: 131 1925AEC65D6C84C7 tf2b_human Transcription initiation factor IIB GTF2B TF2B TFIIB General transcription factor TFIIB S300-II protein protein human Homo sapiens Human MASTSRLDALPRVTCPNHPDAILVEDYRAGDMICPECGLVVGDRVIDVGSEWRTFSNDKATKDPSRVGDSQNPLLSDGDLSTMIGKGTGAASFDEFGNSKYQNRRTMSSSDRAMMNAFKEITTMADRINLPRNIVDRTNNLFKQVYEQKSLKGRANDAIASACLYIACRQEGVPRTFKEICAVSRISKKEIGRCFKLILKALETSVDLITTGDFMSRFCSNLCLPKQVQMAATHIARKAVELDLVPGRSPISVAAAAIYMASQASAEKRTQKEIGDIAGVADVTIRQSYRLIYPRAPDLFPTDFKFDTPVDKLPQL DIP protein Q00403 original sequence version: 119 DIP protein Q00403 original sequence version: 181 9CC7E102526C2722 1433e_human 14-3-3 protein epsilon YWHAE protein protein human Homo sapiens Human MDDREDLVYQAKLAEQAERYDEMVESMKKVAGMDVELTVEERNLLSVAYKNVIGARRASWRIISSIEQKEENKGGEDKLKMIREYRQMVETELKLICCDILDVLDKHLIPAANTGESKVFYYKMKGDYHRYLAEFATGNDRKEAAENSLVAYKAASDIAMTELPPTHPIRLGLALNFSVFYYEILNSPDRACRLAKAAFDDAIAELDTLSEESYKDSTLIMQLLRDNLTLWTSDMQGDGEEQNKEALQDVEDENQ DIP protein P62258 original sequence version: 130 DIP protein P62258 original sequence version: 132 DIP protein P62258 original sequence version: 91 07817CCBD1F75B26 14331_arath 14-3-3-like protein GF14 chi General regulatory factor 1 At4g09000 GRF1 F23J3.30 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MATPGASSARDEFVYMAKLAEQAERYEEMVEFMEKVAKAVDKDELTVEERNLLSVAYKNVIGARRASWRIISSIEQKEESRGNDDHVSLIRDYRSKIETELSDICDGILKLLDTILVPAAASGDSKVFYLKMKGDYHRYLAEFKSGQERKDAAEHTLTAYKAAQDIANSELAPTHPIRLGLALNFSVFYYEILNSPDRACNLAKQAFDEAIAELDTLGEESYKDSTLIMQLLRDNLTLWTSDMQDDVADDIKEAAPAAAKPADEQQS 328FA525D8AE0290 14334_arath 14-3-3-like protein GF14 phi General regulatory factor 4 GRF4 At1g35160 T32G9.30 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAAPPASSSAREEFVYLAKLAEQAERYEEMVEFMEKVAEAVDKDELTVEERNLLSVAYKNVIGARRASWRIISSIEQKEESRGNDDHVTTIRDYRSKIESELSKICDGILKLLDTRLVPASANGDSKVFYLKMKGDYHRYLAEFKTGQERKDAAEHTLTAYKAAQDIANAELAPTHPIRLGLALNFSVFYYEILNSPDRACNLAKQAFDEAIAELDTLGEESYKDSTLIMQLLRDNLTLWTSDMQDESPEEIKEAAAPKPAEEQKEI 18FA010F752CC134 tbp2_arath TATA-box-binding protein 2 TBP2 TATA sequence-binding protein 2 TATA-binding factor 2 TATA-box factor 2 Transcription initiation factor TFIID TBP-2 subunit T5A14.8 At1g55520 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MADQGTEGSQPVDLTKHPSGIVPTLQNIVSTVNLDCKLDLKAIALQARNAEYNPKRFAAVIMRIREPKTTALIFASGKMVCTGAKSEHLSKLAARKYARIVQKLGFPAKFKDFKIQNIVGSCDVKFPIRLEGLAYSHSAFSSYEPELFPGLIYRMKLPKIVLLIFVSGKIVITGAKMREETYTAFENIYPVLREFRKVQQ CFB9596EF8D9CEDD tbp_human TATA-box-binding protein TBP TF2D TFIID GTF2D1 TATA-box factor TATA-binding factor TATA sequence-binding protein Transcription initiation factor TFIID TBP subunit protein protein human Homo sapiens Human MDQNNSLPPYAQGLASPQGAMTPGIPIFSPMMPYGTGLTPQPIQNTNSLSILEEQQRQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQAVAAAAVQQSTSQQATQGTSGQAPQLFHSQTLTTAPLPGTTPLYPSPMTPMTPITPATPASESSGIVPQLQNIVSTVNLGCKLDLKTIALRARNAEYNPKRFAAVIMRIREPRTTALIFSSGKMVCTGAKSEEQSRLAARKYARVVQKLGFPAKFLDFKIQNMVGSCDVKFPIRLEGLVLTHQQFSSYEPELFPGLIYRMIKPRIVLLIFVSGKVVLTGAKVRAEIYEAFENIYPILKGFRKTT DIP protein P20226 original sequence version: 106 DIP protein P20226 original sequence version: 157 DIP protein P20226 original sequence version: 173 DIP protein P20226 original sequence version: 194 DIP protein P20226 original sequence version: 179 DIP protein P20226 original sequence version: 131 A61A578D972B970B 14331_maize 14-3-3-like protein GF14-6 GRF1 protein protein maize Zea mays (Maize) Maize MASAELSREENVYMAKLAEQAERYEEMVEFMEKVAKTVDSEELTVEERNLLSVAYKNVIGARRASWRIISSIEQKEEGRGNEDRVTLIKDYRGKIETELTKICDGILKLLETHLVPSSTAPESKVFYLKMKGDYYRYLAEFKTGAERKDAAENTMVAYKAAQDIALAELAPTHPIRLGLALNFSVFYYEILNSPDRACSLAKQAFDEAISELDTLSEESYKDSTLIMQLLRDNLTLWTSDISEDPAEEIREAPKRDSSEGQ 25EC70725A5F6801 14333_arath 14-3-3-like protein GF14 psi Protein RARE COLD INDUCIBLE 1A RCI1A MXI10.21 At5g38480 GRF3 General regulatory factor 3 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSTREENVYMAKLAEQAERYEEMVEFMEKVAKTVDVEELSVEERNLLSVAYKNVIGARRASWRIISSIEQKEESKGNEDHVAIIKDYRGKIESELSKICDGILNVLEAHLIPSASPAESKVFYLKMKGDYHRYLAEFKAGAERKEAAESTLVAYKSASDIATAELAPTHPIRLGLALNFSVFYYEILNSPDRACSLAKQAFDDAIAELDTLGEESYKDSTLIMQLLRDNLTLWTSDMTDEAGDEIKEASKPDGAE 51E018F146ED2A0F tf2b-grf1 1 8 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 14 unspecified role unspecified role prey prey 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 ecoli Escherichia coli (strain K12) physical association physical association Figure 1A As shown in Figure 1A, maize 14-3-3 was retained on GST-hTFIIB beads but not on the GST control beads. tf2b-grf4 1 8 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region 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 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 ecoli Escherichia coli (strain K12) physical association physical association Figure 1B Figure 2A A series of GST-fused C- or N-terminal deletion mutants of hTFIIB were tested for their ability to interact with the Arabidopsis 14- 3 3 protein AtGF14f. Free 14-3-3 protein was incubated in binding reactions identical to those used for Figure 1 with equal amounts of immobilized GST-hTFIIB deletion mutants. The protein gel blot probed with the anti-14-3-3 polyclonal antibody showed that hTFIIB possesses two regions capable of interacting with the 14-3-3 protein: one is located from amino acids 124 to 202 and the other from amino acids 262 to 292. tf2b-grf3 1 8 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey 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 ecoli Escherichia coli (strain K12) physical association physical association Figure 1B tf2b-grf7 1 6 unspecified role unspecified role prey prey 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 ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 1B tf2b-grf2 1 5 unspecified role unspecified role prey prey 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 ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 1B tf2b-grf1-1 1 10 unspecified role unspecified role prey prey 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 ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 1B grf4-tf2b 2 11 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey region t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q00403:p.[Arg185Glu;Arg193Glu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R E certain certain sequence position certain certain certain sequence position certain R E Fri Jul 22 10:57:59 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg193glu,arg185glu' Wed May 02 15:01:26 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg185glu,arg193glu' Tue Jun 30 19:18:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg185Glu;Arg193Glu]' ecoli Escherichia coli (strain K12) physical association physical association Figure 3 These results suggest that both VP16 and 14-3-3 may use the same or similar mechanisms in binding to hTFIIB. 14_3_3-tbp 2 13 unspecified role unspecified role prey prey region t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P62258:p.[Leu175Ala;Asn176Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain L A certain certain sequence position certain certain certain sequence position certain N A Wed May 02 20:19:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu175ala,asn176ala' Tue Jun 30 17:50:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu175Ala;Asn176Ala]' P62258:p.[Phe177Ala;Ser178Ala] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain F A certain certain sequence position certain certain certain sequence position certain S A Wed May 02 17:01:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe177ala,ser178ala' Tue Jun 30 18:31:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe177Ala;Ser178Ala]' P62258:p.[Val179Ala;Phe180Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain V A certain certain sequence position certain certain certain sequence position certain F A Wed May 02 16:14:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val179ala,phe180ala' Tue Jun 30 17:45:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Val179Ala;Phe180Ala]' ecoli Escherichia coli (strain K12) physical association physical association Figure 5A Figure 7 The GST-h14-3-3 protein was immobilized on glutathione beads and incubated with bacterially expressed hTBP and hTFIIB or with hTAFII32 and hTAFII55 synthesized using a coupled transcription/translation system. h14-3-3 interacted with hTBP 14_3_3-tft2 2 9 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey region t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 5A Figure 6 Immobilized h14-3-3 Interacts with hTBP, hTFIIB, and hTAFII32 but Not with hTAFII55 The authors provide evidence that the seventh helical domain of the 14-3-3 protein is involved in protein interaction (see Figure 6); but do not provide coordinates for the region. I did not attempt to put fuzzy coordinates for the helical region of the 14-3-3 protein shown to be involved in PPi, since the UniProt record does not display this information and prediction tools may produce different results. We have no policy on what to do in this situation so I elected to add a caution. 14_3_3-taf9 3 9 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association Figure 5B The GST�h14-3-3 protein was immobilized on glutathione beads and incubated with bacterially expressed hTBP and hTFIIB or with hTAFII32 and hTAFII55 synthesized using a coupled transcription/translation system. h14-3-3 interacted with hTBP (Figure 5A, cf. lanes 4 and 3), hTFIIB (cf. lanes 2 and 1), and hTAFII32 14_3_3-tbp-1 2 11 unspecified role unspecified role prey prey P46077:p.[Phe186Ala;Ser187Ala] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain F A certain certain sequence position certain certain certain sequence position certain S A Wed May 02 15:55:05 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe186ala,ser187ala' Tue Jun 30 20:13:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe186Ala;Ser187Ala]' P46077:p.[Val188Ala;Phe189Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain F A certain certain sequence position certain certain certain sequence position certain V A Fri Jul 22 10:52:42 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'phe189ala,val188ala' Wed May 02 15:18:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val188ala,phe189ala' Tue Jun 30 18:34:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Val188Ala;Phe189Ala]' region t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P46077:p.[Leu184Ala;Asn185Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain N A certain certain sequence position certain certain certain sequence position certain L A Fri Jul 22 11:02:48 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'asn185ala,leu184ala' Wed May 02 19:04:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu184ala,asn185ala' Tue Jun 30 19:40:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu184Ala;Asn185Ala]' ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 7 14_3_3-tft2-1 2 4 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role prey prey P46077:p.[Val188Ala;Phe189Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain F A certain certain sequence position certain certain certain sequence position certain V A Fri Jul 22 11:11:12 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'phe189ala,val188ala' Wed May 02 17:02:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val188ala,phe189ala' Tue Jun 30 17:52:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Val188Ala;Phe189Ala]' region t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P46077:p.[Phe186Ala;Ser187Ala] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain S A certain certain sequence position certain certain certain sequence position certain F A Fri Jul 22 11:13:05 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'ser187ala,phe186ala' Wed May 02 16:28:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe186ala,ser187ala' Tue Jun 30 18:12:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe186Ala;Ser187Ala]' P46077:p.[Leu184Ala;Asn185Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain L A certain certain sequence position certain certain certain sequence position certain N A Wed May 02 19:01:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu184ala,asn185ala' Tue Jun 30 19:57:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu184Ala;Asn185Ala]' ecoli Escherichia coli (strain K12) physical association physical association Figure 7 As indicated by lanes 5 through 7 in Figure 7, a clear footprint in 14-3-3 binding activity was obtained for both AtTFIIB and AtTBP2. The F186A/S187A mutation nearly eliminated interactions with AtTFIIB and AtTBP2 (cf. lanes 6 and 1), indicating that these two residues are critically important for the interaction. Flanking mutations (L184A/N185A and V188A/F189A) showed partial loss of binding capacity for AtTFIIB and AtTBP2 (cf. lanes 5 and 1 and lanes 7 and 1). Other residues more distal to F186 and S187 appeared to have no involvement in interactions with AtTBP2 or AtTFIIB, because corresponding mutant proteins still showed strong binding (cf. lanes 2, 3, 4, 7, and 8 with lane 1). Mutations for residues Y190 and Y191 were not obtained. As a whole, these results clearly show that the in vitro interactions between AtGF14 and either AtTBP2 or AtTFIIB were highly specific and involved residues in the region from Leu-184 to Phe-189 of helix 7.