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/ The H-NS dimerization domain defines a new fold contributing to DNA recognition. The H-NS dimerization domain defines a new fold contributing to DNA recognition. Nat. Struct. Biol. (1072-8368) 2003 imex curation Bloch V., Yang Y., Margeat E., Chavanieu A., Augé M.T., Robert B., Arold S., Rimsky S., Kochoyan M. The NMR structure was determined using both, labelled and unlabelled, recombinant proteins. Labelling was carried out with 13C and/or 15N. It was finally confirmed using a synthetic peptide ranging from 2-47 aa. The amino acid range was adjusted to 2-47 aa as the n-terminal Methionine was left out of the original strain (see Uniprot history). michel@cbs.univ-montp1.fr 19-Jun-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa weight by bromide molecular weight estimation by bromide staining weight by bromide Bloch V., Yang Y., Margeat E., Chavanieu A., Augé M.T., Robert B., Arold S., Rimsky S., Kochoyan M. The NMR structure was determined using both, labelled and unlabelled, recombinant proteins. Labelling was carried out with 13C and/or 15N. It was finally confirmed using a synthetic peptide ranging from 2-47 aa. The amino acid range was adjusted to 2-47 aa as the n-terminal Methionine was left out of the original strain (see Uniprot history). michel@cbs.univ-montp1.fr Accepted 2013-JUN-13 by MGT 19-Jun-2013: Contacted by IntAct-Help. Nat. Struct. Biol. (1072-8368) 2003 Only protein-protein interactions imex curation The H-NS dimerization domain defines a new fold contributing to DNA recognition. The H-NS dimerization domain defines a new fold contributing to DNA recognition. Nat. Struct. Biol. (1072-8368) 2003 imex curation Bloch V., Yang Y., Margeat E., Chavanieu A., Augé M.T., Robert B., Arold S., Rimsky S., Kochoyan M. The NMR structure was determined using both, labelled and unlabelled, recombinant proteins. Labelling was carried out with 13C and/or 15N. It was finally confirmed using a synthetic peptide ranging from 2-47 aa. The amino acid range was adjusted to 2-47 aa as the n-terminal Methionine was left out of the original strain (see Uniprot history). michel@cbs.univ-montp1.fr 19-Jun-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Bloch V., Yang Y., Margeat E., Chavanieu A., Augé M.T., Robert B., Arold S., Rimsky S., Kochoyan M. The NMR structure was determined using both, labelled and unlabelled, recombinant proteins. Labelling was carried out with 13C and/or 15N. It was finally confirmed using a synthetic peptide ranging from 2-47 aa. The amino acid range was adjusted to 2-47 aa as the n-terminal Methionine was left out of the original strain (see Uniprot history). michel@cbs.univ-montp1.fr Accepted 2013-JUN-13 by MGT 19-Jun-2013: Contacted by IntAct-Help. Nat. Struct. Biol. (1072-8368) 2003 Only protein-protein interactions imex curation The H-NS dimerization domain defines a new fold contributing to DNA recognition. The H-NS dimerization domain defines a new fold contributing to DNA recognition. Nat. Struct. Biol. (1072-8368) 2003 imex curation Bloch V., Yang Y., Margeat E., Chavanieu A., Augé M.T., Robert B., Arold S., Rimsky S., Kochoyan M. The NMR structure was determined using both, labelled and unlabelled, recombinant proteins. Labelling was carried out with 13C and/or 15N. It was finally confirmed using a synthetic peptide ranging from 2-47 aa. The amino acid range was adjusted to 2-47 aa as the n-terminal Methionine was left out of the original strain (see Uniprot history). michel@cbs.univ-montp1.fr 19-Jun-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro crosslink cross-linking study crosslink weight by comassie molecular weight estimation by coomasie staining weight by comassie Bloch V., Yang Y., Margeat E., Chavanieu A., Augé M.T., Robert B., Arold S., Rimsky S., Kochoyan M. The NMR structure was determined using both, labelled and unlabelled, recombinant proteins. Labelling was carried out with 13C and/or 15N. It was finally confirmed using a synthetic peptide ranging from 2-47 aa. The amino acid range was adjusted to 2-47 aa as the n-terminal Methionine was left out of the original strain (see Uniprot history). michel@cbs.univ-montp1.fr Accepted 2013-JUN-13 by MGT 19-Jun-2013: Contacted by IntAct-Help. Nat. Struct. Biol. (1072-8368) 2003 Only protein-protein interactions imex curation The H-NS dimerization domain defines a new fold contributing to DNA recognition. The H-NS dimerization domain defines a new fold contributing to DNA recognition. Nat. Struct. Biol. (1072-8368) 2003 imex curation Bloch V., Yang Y., Margeat E., Chavanieu A., Augé M.T., Robert B., Arold S., Rimsky S., Kochoyan M. The NMR structure was determined using both, labelled and unlabelled, recombinant proteins. Labelling was carried out with 13C and/or 15N. It was finally confirmed using a synthetic peptide ranging from 2-47 aa. The amino acid range was adjusted to 2-47 aa as the n-terminal Methionine was left out of the original strain (see Uniprot history). michel@cbs.univ-montp1.fr 19-Jun-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps predetermined predetermined participant predetermined Bloch V., Yang Y., Margeat E., Chavanieu A., Augé M.T., Robert B., Arold S., Rimsky S., Kochoyan M. The NMR structure was determined using both, labelled and unlabelled, recombinant proteins. Labelling was carried out with 13C and/or 15N. It was finally confirmed using a synthetic peptide ranging from 2-47 aa. The amino acid range was adjusted to 2-47 aa as the n-terminal Methionine was left out of the original strain (see Uniprot history). michel@cbs.univ-montp1.fr Accepted 2013-JUN-13 by MGT 19-Jun-2013: Contacted by IntAct-Help. Nat. Struct. Biol. (1072-8368) 2003 Only protein-protein interactions imex curation hns_ecoli DNA-binding protein H-NS hns bglY cur drdX hnsA msyA osmZ pilG topS b1237 Histone-like protein HLP-II Protein H1 Protein B1 JW1225 Heat-stable nucleoid-structuring protein protein protein ecoli Escherichia coli (strain K12) MSEALKILNNIRTLRAQARECTLETLEEMLEKLEVVVNERREEESAAAAEVEERTRKLQQYREMLIADGIDPNELLNSLAAVKSGTKAKRAQRPAKYSYVDENGETKTWTGQGRTPAVIKKAMDEQGKSLDDFLIKQ DIP protein P0ACF8 original sequence version: 44 E628184AC7C86F49 curved dna curved DNA fragment 220bp containing this sequence ds dna double stranded deoxyribonucleic acid ds DNA ds dna chemical synthesis Chemical synthesis (Chemical synthesis) TCTCTAAAAAATATATAAAAATCTCTAAAAAATATATAAAAATCTCTAAAAAATATATAAAAA hns-1 2 5 unspecified role unspecified role neutral component neutral component region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 13c label 13c label C13 13C C13 13C undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction Fig. 2 The experiment was carried out with 2 constructs: a) a recombinant protein with a n-terminal his-tag, expressed in ecoli and b) a synthetic peptide. hns-dimer 4 5 unspecified role unspecified role neutral component neutral component n-terminal alexa 488 label alexa 488 label n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Fig. 1 The dimer occurs predominantly at low concentrations. hns-oligomer 4 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal alexa 488 label alexa 488 label n-term range n-terminal range n-term range n-term range n-terminal range n-term range chemical synthesis Chemical synthesis (Chemical synthesis) 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal alexa 488 label alexa 488 label n-term range n-terminal range n-term range n-term range n-terminal range n-term range chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Fig. 1 The oligomers form at higher concentrations. hns-dimer 3 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Fig. 1 curved_dna-hns 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 n-term range n-terminal range n-term range n-term range n-terminal range n-term range P0ACF8:p.[Arg12Glu;Arg15Ala] 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 A Wed May 02 17:02:05 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg12glu,arg15ala' Tue Jun 30 18:22:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg12Glu;Arg15Ala]' ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role bait bait chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Fig. 4b DNA binding is concentration dependent: A molar ratio of at least 4:1 protein:DNA is required for binding. The double mutant recognizes DNA with a slightly weaker affinity than the wild type molecule, and it has lost its ability to preferentially recognize intrinsically curved DNA sequences.