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/ Characterization of SARS-CoV-2 N protein reveals multiple functional consequences of the C-terminal domain Characterization of SARS-CoV-2 N protein reveals multiple functional consequences of the C-terminal domain iScience 2021 imex curation Wu C., Qavi AJ., Hachim A., Kavian N., Cole AR., Moyle AB., Wagner ND., Sweeney-Gibbons J., Rohrs HW., Gross ML., Peiris JSM., Basler CF., Farnsworth CW., Valkenburg SA., Amarasinghe GK., Leung DW. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus most results from Figure 4 cannot be annotated as the RNA is of bacterial origin, sequence unknown. cwfarnsworth@wustl.edu,gamarasinghe@wustl.edu,dwleung@wustl.edu Only protein-protein interactions imex curation in vitro In vitro h1-h2 ms mass spectrometry study of hydrogen/deuterium exchange h1-h2 ms sequence tag sequence tag identification sequence tag Wu C., Qavi AJ., Hachim A., Kavian N., Cole AR., Moyle AB., Wagner ND., Sweeney-Gibbons J., Rohrs HW., Gross ML., Peiris JSM., Basler CF., Farnsworth CW., Valkenburg SA., Amarasinghe GK., Leung DW. Virus - Publications including interactions involving viral proteins most results from Figure 4 cannot be annotated as the RNA is of bacterial origin, sequence unknown. cwfarnsworth@wustl.edu,gamarasinghe@wustl.edu,dwleung@wustl.edu Accepted 2021-JUL-19 AT 07:56 BST AT 07:56 BST by ORCHARD iScience 2021 Only protein-protein interactions imex curation imex curation Characterization of SARS-CoV-2 N protein reveals multiple functional consequences of the C-terminal domain Characterization of SARS-CoV-2 N protein reveals multiple functional consequences of the C-terminal domain iScience 2021 imex curation Wu C., Qavi AJ., Hachim A., Kavian N., Cole AR., Moyle AB., Wagner ND., Sweeney-Gibbons J., Rohrs HW., Gross ML., Peiris JSM., Basler CF., Farnsworth CW., Valkenburg SA., Amarasinghe GK., Leung DW. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus most results from Figure 4 cannot be annotated as the RNA is of bacterial origin, sequence unknown. cwfarnsworth@wustl.edu,gamarasinghe@wustl.edu,dwleung@wustl.edu Only protein-protein interactions imex curation ecoli Escherichia coli (strain K12) dls dynamic light scattering dls predetermined predetermined participant predetermined Wu C., Qavi AJ., Hachim A., Kavian N., Cole AR., Moyle AB., Wagner ND., Sweeney-Gibbons J., Rohrs HW., Gross ML., Peiris JSM., Basler CF., Farnsworth CW., Valkenburg SA., Amarasinghe GK., Leung DW. Virus - Publications including interactions involving viral proteins most results from Figure 4 cannot be annotated as the RNA is of bacterial origin, sequence unknown. cwfarnsworth@wustl.edu,gamarasinghe@wustl.edu,dwleung@wustl.edu Accepted 2021-JUL-19 AT 07:56 BST AT 07:56 BST by ORCHARD iScience 2021 Only protein-protein interactions imex curation imex curation Characterization of SARS-CoV-2 N protein reveals multiple functional consequences of the C-terminal domain Characterization of SARS-CoV-2 N protein reveals multiple functional consequences of the C-terminal domain iScience 2021 imex curation Wu C., Qavi AJ., Hachim A., Kavian N., Cole AR., Moyle AB., Wagner ND., Sweeney-Gibbons J., Rohrs HW., Gross ML., Peiris JSM., Basler CF., Farnsworth CW., Valkenburg SA., Amarasinghe GK., Leung DW. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus most results from Figure 4 cannot be annotated as the RNA is of bacterial origin, sequence unknown. cwfarnsworth@wustl.edu,gamarasinghe@wustl.edu,dwleung@wustl.edu Only protein-protein interactions imex curation in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps tag fluorescence tag visualisation by fluorescence tag fluorescence Wu C., Qavi AJ., Hachim A., Kavian N., Cole AR., Moyle AB., Wagner ND., Sweeney-Gibbons J., Rohrs HW., Gross ML., Peiris JSM., Basler CF., Farnsworth CW., Valkenburg SA., Amarasinghe GK., Leung DW. Virus - Publications including interactions involving viral proteins most results from Figure 4 cannot be annotated as the RNA is of bacterial origin, sequence unknown. cwfarnsworth@wustl.edu,gamarasinghe@wustl.edu,dwleung@wustl.edu Accepted 2021-JUL-19 AT 07:56 BST AT 07:56 BST by ORCHARD iScience 2021 Only protein-protein interactions imex curation imex curation ncap_sars2 Nucleoprotein N Nucleocapsid protein protein protein SARS-CoV-2 SARS-CoV-2 SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 2019-nCoV MSDNGPQNQRNAPRITFGGPSDSTGSNQNGERSGARSKQRRPQGLPNNTASWFTALTQHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGKMKDLSPRWYFYYLGTGPEAGLPYGANKDGIIWVATEGALNTPKDHIGTRNPANNAAIVLQLPQGTTLPKGFYAEGSRGGSQASSRSSSRSRNSSRNSTPGSSRGTSPARMAGNGGDAALALLLLDRLNQLESKMSGKGQQQQGQTVTKKSAAEASKKPRQKRTATKAYNVTQAFGRRGPEQTQGNFGDQELIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDDKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKADETQALPQRQKKQQTVTLLPAADLDDFSKQLQQSMSSADSTQA 20-nt ssrna rna ribonucleic acid RNA rna chemical synthesis Chemical synthesis (Chemical synthesis) UUUCACCUCCCUUUCAGUUU stem-loop-rna rna ribonucleic acid RNA rna chemical synthesis Chemical synthesis (Chemical synthesis) GGAAGAUUAAUAAUUUUCC n-n-80 2 4 predetermined predetermined participant predetermined sequence tag sequence tag identification sequence tag experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified 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 NTD-LKR-CTD region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 4 predetermined predetermined participant predetermined sequence tag sequence tag identification sequence tag experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified 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 NTD-LKR-CTD region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction 1C, Supp. figure 2 There are two major oligomeric species for full-length N (NWT; 46 kDa), with hydrodynamic radii (Rh) of 8.9 nm and 450 nm (Figures 1C and 1D), suggesting the presence of different oligomeric forms of the protein. For truncated NNTD-LKR-CTD construct results show two major species that are similar to N-WT. However, both NNTD-LKR-CTD populations display reduced polydispersity. n-n-81 2 4 predetermined predetermined participant predetermined sequence tag sequence tag identification sequence tag experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified 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 NTD-LKR region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain CTD region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction 1C, Supp. figure 2 N CTD is a dimer in solution;N NTD-LKR results in a single peak representing a dimeric species. 20_nt ssrna-n-1 3 5 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 chemical synthesis Chemical synthesis (Chemical synthesis) 4 unspecified role unspecified role neutral component neutral component NTD-LKR-CTD region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 CTD sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P0DTC9:p.[Ser176Asp;Ser188Asp;Ser206Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain S D certain certain sequence position certain certain certain sequence position certain S D certain certain sequence position certain certain certain sequence position certain S D Tue Oct 05 18:38:07 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser176Asp;Ser188Asp;Ser206Asp]' LKR-CTD region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain NTD-LKR-CTD -Carm-region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain NTD-LKR region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain NTD sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P0DTC9:p.[Ser188Asp;Ser206Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain S D certain certain sequence position certain certain certain sequence position certain S D Tue Oct 05 19:13:50 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser188Asp;Ser206Asp]' ecoli Escherichia coli (strain K12) direct interaction direct interaction 2, 4E, Supp. figure 3,5 Figure 4E gives these Kd values: For the S176D/S188D/S206D mutant:- N=0.015uM (WT =0.007uM); N-NTD-LKR=1.1uM (WT=0.5uM). Supp. figure 5A gives these Kd values:- For the S176D/S188D/S206D mutant:- N=0.023uM (WT =0.007uM); For the N S188D/S206D mutant= 0.015 uM. No Kd value is given for the LKR-CTD region. 20_nt ssrna-n-2 3 6 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 chemical synthesis Chemical synthesis (Chemical synthesis) 4 unspecified role unspecified role neutral component neutral component NTD-LKR-CTD region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 CTD sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain NTD-LKR-CTD -Carm-region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain NTD-LKR region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain NTD sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain LKR+CTD region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P0DTC9:p.[Ser176Asp;Ser188Asp;Ser206Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain S D certain certain sequence position certain certain certain sequence position certain S D certain certain sequence position certain certain certain sequence position certain S D Tue Oct 05 18:38:12 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser176Asp;Ser188Asp;Ser206Asp]' ecoli Escherichia coli (strain K12) direct interaction direct interaction 2, Supp. figure 3, 5 Supp. Figure 5 give Kd for N NTD-LKR as 3.0uM, while Figure 2 gives a value of 2.5uM. 20_nt ssrna-n-3 1 4 unspecified role unspecified role neutral component neutral component P0DTC9:p.[Ser176Asp;Ser188Asp;Ser206Asp] mutation mutation certain certain sequence position certain certain certain sequence position certain S D certain certain sequence position certain certain certain sequence position certain S D certain certain sequence position certain certain certain sequence position certain S D Tue Oct 05 19:05:00 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser176Asp;Ser188Asp;Ser206Asp]' 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 region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain NTD-LKR 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) 5 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 chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction 3, Supp. figure 4