UniProt UniProt http://www.uniprot.org The c-di-GMP binding protein YcgR controls flagellar motor direction and speed to affect chemotaxis by a "backstop brake" mechanism. The c-di-GMP binding protein YcgR controls flagellar motor direction and speed to affect chemotaxis by a "backstop brake" mechanism. Mol. Cell (1097-2765) 2010 imex curation Paul K., Nieto V., Carlquist W.C., Blair D.F., Harshey R.M. rasika@uts.cc.utexas.edu,blair@bioscience.utah.edu 21-Dec-2011: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro crosslink cross-linking study crosslink anti tag western anti tag western blot anti tag western Paul K., Nieto V., Carlquist W.C., Blair D.F., Harshey R.M. blair@bioscience.utah.edu rasika@uts.cc.utexas.edu rasika@uts.cc.utexas.edu,blair@bioscience.utah.edu Accepted 2011-SEP-30 by ORCHARD 21-Dec-2011: Contacted by JYOTI. Mol. Cell (1097-2765) 2010 Only protein-protein interactions imex curation The c-di-GMP binding protein YcgR controls flagellar motor direction and speed to affect chemotaxis by a "backstop brake" mechanism. The c-di-GMP binding protein YcgR controls flagellar motor direction and speed to affect chemotaxis by a "backstop brake" mechanism. Mol. Cell (1097-2765) 2010 imex curation Paul K., Nieto V., Carlquist W.C., Blair D.F., Harshey R.M. rasika@uts.cc.utexas.edu,blair@bioscience.utah.edu 21-Dec-2011: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Paul K., Nieto V., Carlquist W.C., Blair D.F., Harshey R.M. blair@bioscience.utah.edu rasika@uts.cc.utexas.edu rasika@uts.cc.utexas.edu,blair@bioscience.utah.edu Accepted 2011-SEP-30 by ORCHARD Mol. Cell (1097-2765) 2010 Only protein-protein interactions imex curation flim_ecoli Flagellar motor switch protein FliM fla AII fla QII b1945 fliM cheC2 JW1929 protein protein ecoli Escherichia coli (strain K12) MGDSILSQAEIDALLNGDSEVKDEPTASVSGESDIRPYDPNTQRRVVRERLQALEIINERFARHFRMGLFNLLRRSPDITVGAIRIQPYHEFARNLPVPTNLNLIHLKPLRGTGLVVFSPSLVFIAVDNLFGGDGRFPTKVEGREFTHTEQRVINRMLKLALEGYSDAWKAINPLEVEYVRSEMQVKFTNITTSPNDIVVNTPFHVEIGNLTGEFNICLPFSMIEPLRELLVNPPLENSRNEDQNWRDNLVRQVQHSQLELVANFADISLRLSQILKLNPGDVLPIEKPDRIIAHVDGVPVLTSQYGTLNGQYALRIEHLINPILNSLNEEQPK DIP protein P06974 original sequence version: null 05DC012B9364BFD6 flig_ecoli Flagellar motor switch protein FliG fliG fla AII.2 fla BII b1939 JW1923 protein protein ecoli Escherichia coli (strain K12) MSNLTGTDKSVILLMTIGEDRAAEVFKHLSQREVQTLSAAMANVTQISNKQLTDVLAEFEQEAEQFAALNINANDYLRSVLVKALGEERAASLLEDILETRDTASGIETLNFMEPQSAADLIRDEHPQIIATILVHLKRAQAADILALFDERLRHDVMLRIATFGGVQPAALAELTEVLNGLLDGQNLKRSKMGGVRTAAEIINLMKTQQEEAVITAVREFDGELAQKIIDEMFLFENLVDVDDRSIQRLLQEVDSESLLIALKGAEQPLREKFLRNMSQRAADILRDDLANRGPVRLSQVENEQKAILLIVRRLAETGEMVIGSGEDTYV DIP protein P0ABZ1 original sequence version: 85 E5CB4C0415C6E178 ycgr_ecoli Flagellar brake protein YcgR ycgR b1194 Cyclic di-GMP binding protein YcgR JW1183 protein protein ecoli Escherichia coli (strain K12) MSHYHEQFLKQNPLAVLGVLRDLHKAAIPLRLSWNGGQLISKLLAITPDKLVLDFGSQAEDNIAVLKAQHITITAETQGAKVEFTVEQLQQSEYLQLPAFITVPPPTLWFVQRRRYFRISAPLHPPYFCQTKLADNSTLRFRLYDLSLGGMGALLETAKPAELQEGMRFAQIEVNMGQWGVFHFDAQLISISERKVIDGKNETITTPRLSFRFLNVSPTVERQLQRIIFSLEREAREKADKVRD DIP protein P76010 original sequence version: null 85D76B51DAC7BAB9 ycgr-flig-flim 2 3 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role bait bait n-terminal tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-terminal n-terminal position amino-terminus n-terminus n-term n-term amino-terminus n-terminus n-terminal n-terminal n-terminal position amino-terminus n-terminus n-term n-term amino-terminus n-terminus n-terminal ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) association association Fig 2A, 2B, 2C Binding is improved in the presence of 5 uM c-di-GMP ycgr-flim 2 5 unspecified role unspecified role bait bait n-terminal tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag predetermined featur predetermined feature predetermined featur certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 3 unspecified role unspecified role prey prey P06974:p.Leu160Glu mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L E Wed May 02 17:42:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu160glu' Tue Jun 30 17:18:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu160Glu' Figure legend says L160D but paper discusses L160E. P06974:p.Arg156Glu mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R E Wed May 02 18:55:26 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg156glu' Tue Jun 30 19:14:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg156Glu' P06974:p.Val153Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain V D Wed May 02 20:11:26 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val153asp' Tue Jun 30 16:04:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val153Asp' P06974:p.Asn155Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain N E Wed May 02 18:43:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn155glu' Tue Jun 30 19:12:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn155Glu' P06974:p.Asp167Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain D R Wed May 02 16:44:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp167arg' Tue Jun 30 18:28:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp167Arg' P06974:p.Glu176Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain E R Wed May 02 16:41:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu176arg' Tue Jun 30 18:07:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu176Arg' ecoli Escherichia coli (strain K12) physical association physical association Fig 3A Binding is improved in the presence of 5 uM c-di-GMP ycgr-flig 2 5 unspecified role unspecified role bait bait n-terminal tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag predetermined featur predetermined feature predetermined featur certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey P0ABZ1:p.Asn292Trp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain N W Wed May 02 19:24:41 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn292trp' Tue Jun 30 17:31:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn292Trp' P0ABZ1:p.Gln252Trp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Q W Wed May 02 17:36:03 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln252trp' Tue Jun 30 18:02:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln252Trp' P0ABZ1:p.Pro295Trp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain P W Wed May 02 14:32:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro295trp' Tue Jun 30 18:57:22 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro295Trp' ecoli Escherichia coli (strain K12) physical association physical association Fig 3C Binding is improved in the presence of 5 uM c-di-GMP ycgr-flim-1 2 5 unspecified role unspecified role bait bait n-terminal tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-terminal n-terminal position amino-terminus n-terminus n-term n-term amino-terminus n-terminus n-terminal n-terminal n-terminal position amino-terminus n-terminus n-term n-term amino-terminus n-terminus n-terminal P76010:p.Gln223Pro mutation disrupting mutation disrupting interaction mutation disrupting polyclonal western polyclonal antibody western blot certain certain sequence position certain certain certain sequence position certain Q P Wed May 02 19:59:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln223pro' Tue Jun 30 18:06:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln223Pro' P76010:p.Ile227Trp mutation disrupting mutation disrupting interaction mutation disrupting polyclonal western polyclonal antibody western blot certain certain sequence position certain certain certain sequence position certain I W Wed May 02 16:20:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile227trp' Tue Jun 30 18:32:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile227Trp' c-terminal tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-terminal c-terminal position c-term carboxy-terminus c-terminus c-term c-terminus carboxy-terminus c-terminal c-terminal c-terminal position c-term carboxy-terminus c-terminus c-term c-terminus carboxy-terminus c-terminal ecoli Escherichia coli (strain K12) 3 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) physical association physical association Fig 5B Wt control not show in Fig 5B, however pg 133 states that another YcgR mutation (H183D) is as strong as wild-type, so used that as +ve control for the FliM-YcgR interaction. Binding requires 5 uM c-di-GMP. flig-ycgr 2 5 unspecified role unspecified role bait bait P76010:p.Asn62Trp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing polyclonal western polyclonal antibody western blot certain certain sequence position certain certain certain sequence position certain N W Wed May 02 16:17:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn62trp' Tue Jun 30 18:58:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn62Trp' n-terminal tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag anti tag western anti tag western blot anti tag western certain certain sequence position certain certain certain sequence position certain c-terminal tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag anti tag western anti tag western blot anti tag western c-terminal c-terminal position c-term carboxy-terminus c-terminus c-term c-terminus carboxy-terminus c-terminal c-terminal c-terminal position c-term carboxy-terminus c-terminus c-term c-terminus carboxy-terminus c-terminal P76010:p.Lys81Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing polyclonal western polyclonal antibody western blot certain certain sequence position certain certain certain sequence position certain K D Wed May 02 18:10:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys81asp' Tue Jun 30 17:21:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys81Asp' Paper discussed K82W, but residue 82 is Val, residue 81 is Lys. Figure 5B legend indicates K81D. ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) physical association physical association Fig 5B flig-flig-2 1 4 unspecified role unspecified role putative self putative self c-terminal tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag certain certain sequence position certain certain certain sequence position certain P0ABZ1:p.Arg297Cys mutation mutation certain certain sequence position certain certain certain sequence position certain R C Wed May 02 19:44:17 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg297cys' Tue Jun 30 17:23:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg297Cys' ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role putative self putative self c-terminal tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag certain certain sequence position certain certain certain sequence position certain P0ABZ1:p.Arg297Cys mutation mutation certain certain sequence position certain certain certain sequence position certain R C Wed May 02 20:27:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg297cys' Tue Jun 30 15:54:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg297Cys' ecoli Escherichia coli (strain K12) disulfide bond disulfide bond disulfide bridge SS-bond Fig 6B Results are not fully commented, only a putative monomer and dimer are shown