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/ A hand-off mechanism for primosome assembly in replication restart. A hand-off mechanism for primosome assembly in replication restart. Mol. Cell (1097-2765) 2007 imex curation Lopper M., Boonsombat R., Sandler S.J., Keck J.L. jlkeck@wisc.edu 27-Mar-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps tag fluorescence tag visualisation by fluorescence tag fluorescence Lopper M., Boonsombat R., Sandler S.J., Keck J.L. jlkeck@wisc.edu Accepted 2013-MAR-14 by ORCHARD 27-Mar-2013: Contacted by IntAct-Help. Mol. Cell (1097-2765) 2007 Only protein-protein interactions imex curation A hand-off mechanism for primosome assembly in replication restart. A hand-off mechanism for primosome assembly in replication restart. Mol. Cell (1097-2765) 2007 imex curation Lopper M., Boonsombat R., Sandler S.J., Keck J.L. jlkeck@wisc.edu 27-Mar-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 Lopper M., Boonsombat R., Sandler S.J., Keck J.L. jlkeck@wisc.edu Accepted 2013-MAR-14 by ORCHARD 27-Mar-2013: Contacted by IntAct-Help. Mol. Cell (1097-2765) 2007 Only protein-protein interactions imex curation dnat_ecoli Primosomal protein 1 dnaT b4362 JW4326 Primosomal protein I protein protein ecoli Escherichia coli (strain K12) MSSRVLTPDVVGIDALVHDHQTVLAKAEGGVVAVFANNAPAFYAVTPARLAELLALEEKLARPGSDVALDDQLYQEPQAAPVAVPMGKFAMYPDWQPDADFIRLAALWGVALREPVTTEELASFIAYWQAEGKVFHHVQWQQKLARSLQIGRASNGGLPKRDVNTVSEPDSQIPPGFRG 8DE048A3B2A6235B 8DE048A3B2A6235B prib_ecoli Primosomal replication protein N priB b4201 JW4159 protein protein ecoli Escherichia coli (strain K12) MTNRLVLSGTVCRAPLRKVSPSGIPHCQFVLEHRSVQEEAGFHRQAWCQMPVIVSGHENQAITHSITVGSRITVQGFISCHKAKNGLSKMVLHAEQIELIDSGD DIP protein P07013 original sequence version: 92 0D52F9D68193B4AC pria_ecoli Primosomal protein N' priA b3935 Replication factor Y JW3906 ATP-dependent helicase PriA protein protein ecoli Escherichia coli (strain K12) MPVAHVALPVPLPRTFDYLLPEGMTVKAGCRVRVPFGKQQERIGIVVSVSDASELPLNELKAVVEVLDSEPVFTHSVWRLLLWAADYYHHPIGDVLFHALPILLRQGRPAANAPMWYWFATEQGQAVDLNSLKRSPKQQQALAALRQGKIWRDQVATLEFNDAALQALRKKGLCDLASETPEFSDWRTNYAVSGERLRLNTEQATAVGAIHSAADTFSAWLLAGVTGSGKTEVYLSVLENVLAQGKQALVMVPEIGLTPQTIARFRERFNAPVEVLHSGLNDSERLSAWLKAKNGEAAIVIGTRSALFTPFKNLGVIVIDEEHDSSYKQQEGWRYHARDLAVYRAHSEQIPIILGSATPALETLCNVQQKKYRLLRLTRRAGNARPAIQHVLDLKGQKVQAGLAPALITRMRQHLQADNQVILFLNRRGFAPALLCHDCGWIAECPRCDHYYTLHQAQHHLRCHHCDSQRPVPRQCPSCGSTHLVPVGLGTEQLEQTLAPLFPGVPISRIDRDTTSRKGALEQQLAEVHRGGARILIGTQMLAKGHHFPDVTLVALLDVDGALFSADFRSAERFAQLYTQVAGRAGRAGKQGEVVLQTHHPEHPLLQTLLYKGYDAFAEQALAERRMMQLPPWTSHVIVRAEDHNNQHAPLFLQQLRNLILSSPLADEKLWVLGPVPALAPKRGGRWRWQILLQHPSRVRLQHIINGTLALINTIPDSRKVKWVLDVDPIEG DIP protein P17888 original sequence version: null AB6C90F43042A363 ssdna-1 Single-stranded deoxyribonucleic acid ss dna single stranded deoxyribonucleic acid ss DNA ss dna ecoli Escherichia coli (strain K12) pria-prib-ssdna 1 4 unspecified role unspecified role neutral component neutral component n-terminal 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 region fitc labelled fluorescein isothiocyanate label fluorescein isothiocyanate labbeled FITC labelled FITC labelled fluorescein isothiocyanate labbeled fitc labelled undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 5 unspecified role unspecified role neutral component neutral component n-terminal 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 2a, Tab. 1 pria-prib-prib-ssdna 1 6 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 4 competitor competitor competitor neutral component neutral component n-terminal 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 P07013:p.Arg44Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R A Wed May 02 17:06:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg44ala' Tue Jun 30 17:31:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg44Ala' P07013:p.[Trp47Ala;Lys82Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain W A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 19:58:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp47ala,lys82ala' Tue Jun 30 18:42:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Trp47Ala;Lys82Ala]' P07013:p.Glu39Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain E A Wed May 02 14:33:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu39ala' Tue Jun 30 17:31:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu39Ala' ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component n-terminal 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 ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role neutral component neutral component region fitc labelled fluorescein isothiocyanate label fluorescein isothiocyanate labbeled FITC labelled FITC labelled fluorescein isothiocyanate labbeled fitc labelled undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 2b, Tab. 1 pria-prib 1 5 unspecified role unspecified role neutral component neutral component n-terminal 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 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) 4 unspecified role unspecified role neutral component neutral component region fitc labelled fluorescein isothiocyanate label fluorescein isothiocyanate labbeled FITC labelled FITC labelled fluorescein isothiocyanate labbeled fitc labelled undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 2a prib-ssdna-1 1 4 unspecified role unspecified role neutral component neutral component P07013:p.Arg44Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R A Wed May 02 18:28:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg44ala' Tue Jun 30 18:33:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg44Ala' P07013:p.Lys82Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K A Wed May 02 16:31:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys82ala' Tue Jun 30 17:46:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys82Ala' n-terminal 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 P07013:p.[Trp47Ala;Lys82Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain W A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 19:15:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp47ala,lys82ala' Tue Jun 30 19:22:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Trp47Ala;Lys82Ala]' ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component region fluorescein fluorescein label fluorescein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association Fig. 2c, Tab. 1 dnat-prib 1 3 unspecified role unspecified role neutral component neutral component region fitc labelled fluorescein isothiocyanate label fluorescein isothiocyanate labbeled FITC labelled FITC labelled fluorescein isothiocyanate labbeled fitc labelled undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal 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 ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role neutral component neutral component P07013:p.[Trp47Ala;Lys82Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain W A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 19:09:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp47ala,lys82ala' Tue Jun 30 18:46:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Trp47Ala;Lys82Ala]' P07013:p.Gln45Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain Q A Wed May 02 19:16:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln45ala' Tue Jun 30 18:44:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln45Ala' n-terminal 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 3a, Tab 1 While an increase in fluorescence anisotropy of FITC-DnaT following addition of PriB was observed, the interaction requires high concentrations of PriB and is not saturable, precluding KD,app determination. Thus, while a direct, physical interaction exists between PriB and DnaT, it needs additional factors. dnat-prib-ssdna 1 4 unspecified role unspecified role neutral component neutral component P07013:p.Lys84Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K A Wed May 02 15:53:30 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys84ala' Tue Jun 30 17:39:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys84Ala' P07013:p.[Trp47Ala;Lys82Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain W A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 19:47:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp47ala,lys82ala' Tue Jun 30 19:26:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Trp47Ala;Lys82Ala]' n-terminal 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 P07013:p.Lys89Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K A Wed May 02 20:11:32 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys89ala' Tue Jun 30 19:46:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys89Ala' P07013:p.Arg44Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R A Wed May 02 15:19:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg44ala' Tue Jun 30 16:50:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg44Ala' P07013:p.Lys82Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K A Wed May 02 15:54:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys82ala' Tue Jun 30 19:27:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys82Ala' P07013:p.Phe42Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain F A Wed May 02 15:10:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe42ala' Tue Jun 30 16:37:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe42Ala' P07013:p.Glu39Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain E A Wed May 02 15:10:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu39ala' Tue Jun 30 19:21:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu39Ala' P07013:p.Trp47Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain W A Wed May 02 15:31:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp47ala' Tue Jun 30 18:51:22 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp47Ala' ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component region fluorescein fluorescein label fluorescein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 3 unspecified role unspecified role neutral component neutral component n-terminal 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 3b, Tab. 1 PriB has a higher affinity for DnaT than ssDNA. Therefore, DnaT displaced PriB from ssDNA and bound to it reducing the anisotrophy of labelled ssDNA. If DnaT fails to bind a PriB mutant, the PriB mutant remains bound to the ssDNA. dnat-pria-prib-ssdna 1 4 unspecified role unspecified role neutral component neutral component P07013:p.Arg44Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R A Wed May 02 15:12:35 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg44ala' Tue Jun 30 18:31:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg44Ala' P07013:p.Glu39Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain E A Wed May 02 17:18:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu39ala' Wed May 02 19:11:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu39ala' Tue Jun 30 19:31:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu39Ala' n-terminal 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 ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component n-terminal 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 ecoli Escherichia coli (strain K12) 3 unspecified role unspecified role neutral component neutral component n-terminal 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 region fitc labelled fluorescein isothiocyanate label fluorescein isothiocyanate labbeled FITC labelled FITC labelled fluorescein isothiocyanate labbeled fitc labelled undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association Fig. 5a prib-prib 2 4 unspecified role unspecified role unspecified role unspecified role n-terminal 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 P07013:p.Glu39Ala mutation mutation certain certain sequence position certain certain certain sequence position certain E A Wed May 02 20:29:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu39ala' Tue Jun 30 16:03:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu39Ala' ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. S2 Rwork = 0.25; Rfree = 0.28 2.25A