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/ Insights into negative modulation of E. coli replication initiation from the structure of SeqA-hemimethylated DNA complex. Insights into negative modulation of E. coli replication initiation from the structure of SeqA-hemimethylated DNA complex. Nat. Struct. Biol. (1072-8368) 2002 imex curation Guarné A., Zhao Q., Ghirlando R., Yang W. wei.yang@nih.gov 26-Jun-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Guarné A., Zhao Q., Ghirlando R., Yang W. wei.yang@nih.gov Accepted 2013-JUN-21 by PPORRAS 26-Jun-2013: Contacted by IntAct-Help. Nat. Struct. Biol. (1072-8368) 2002 Only protein-protein interactions imex curation Insights into negative modulation of E. coli replication initiation from the structure of SeqA-hemimethylated DNA complex. Insights into negative modulation of E. coli replication initiation from the structure of SeqA-hemimethylated DNA complex. Nat. Struct. Biol. (1072-8368) 2002 imex curation Guarné A., Zhao Q., Ghirlando R., Yang W. wei.yang@nih.gov 26-Jun-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Guarné A., Zhao Q., Ghirlando R., Yang W. wei.yang@nih.gov Accepted 2013-JUN-21 by PPORRAS 26-Jun-2013: Contacted by IntAct-Help. Nat. Struct. Biol. (1072-8368) 2002 Only protein-protein interactions imex curation Insights into negative modulation of E. coli replication initiation from the structure of SeqA-hemimethylated DNA complex. Insights into negative modulation of E. coli replication initiation from the structure of SeqA-hemimethylated DNA complex. Nat. Struct. Biol. (1072-8368) 2002 imex curation Guarné A., Zhao Q., Ghirlando R., Yang W. wei.yang@nih.gov 26-Jun-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Guarné A., Zhao Q., Ghirlando R., Yang W. wei.yang@nih.gov Accepted 2013-JUN-21 by PPORRAS 26-Jun-2013: Contacted by IntAct-Help. Nat. Struct. Biol. (1072-8368) 2002 Only protein-protein interactions imex curation Insights into negative modulation of E. coli replication initiation from the structure of SeqA-hemimethylated DNA complex. Insights into negative modulation of E. coli replication initiation from the structure of SeqA-hemimethylated DNA complex. Nat. Struct. Biol. (1072-8368) 2002 imex curation Guarné A., Zhao Q., Ghirlando R., Yang W. wei.yang@nih.gov 26-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 Guarné A., Zhao Q., Ghirlando R., Yang W. Crosslinking agent was BS3 [bis(sulfosuccinimidyl)suberate] wei.yang@nih.gov Accepted 2013-JUN-21 by PPORRAS 26-Jun-2013: Contacted by IntAct-Help. Nat. Struct. Biol. (1072-8368) 2002 Only protein-protein interactions imex curation seqa_ecoli Negative modulator of initiation of replication seqA b0687 JW0674 protein protein ecoli Escherichia coli (strain K12) MKTIEVDDELYSYIASHTKHIGESASDILRRMLKFSAASQPAAPVTKEVRVASPAIVEAKPVKTIKDKVRAMRELLLSDEYAEQKRAVNRFMLLLSTLYSLDAQAFAEATESLHGRTRVYFAADEQTLLKNGNQTKPKHVPGTPYWVITNTNTGRKCSMIEHIMQSMQFPAELIEKVCGTI 37D9078CA3D7229B 37D9078CA3D7229B hemimeth_dna_rev hemimethylated DNA from ecoli oriC reverse ss dna single stranded deoxyribonucleic acid ss DNA ss dna chemical synthesis Chemical synthesis (Chemical synthesis) AGTCGATCGGTG hemimeth_dna_forw hemimethylated DNA from ecoli oriC forward ss dna single stranded deoxyribonucleic acid ss DNA ss dna chemical synthesis Chemical synthesis (Chemical synthesis) CACCGATCGACT seqa-1 1 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 1b Tetramer present when SeqA in low concentration or in 5% (v/v) isopropanol. Otherwise forms larger homo-multimers. seqa-1 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 1c dnaa-hemimeth 3 5 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component dna binding binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 7 unspecified role unspecified role neutral component neutral component dna binding binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 17 19 physical association physical association Fig. 1d Reverse DNA oligo is methylated on Adenine6. dnaa-hemimeth-2 2 6 unspecified role unspecified role neutral component neutral component dna binding binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 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 region se(s)met L-selenomethionine Delta:S(-1)Se(1) Se(S)Met Selenium replaces sulphur semetm undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component dna binding binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 27 22 physical association physical association Fig. 2 Rwork = 0.25; Rfree = 0.28 2.65 A Reverse DNA oligo is methylated on Adenine6.