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 nsp9 replicase protein of SARS-coronavirus, structure and functional insights. The nsp9 replicase protein of SARS-coronavirus, structure and functional insights. Structure (London, England : 1993) 2004 imex curation Sutton G., Fry E., Carter L., Sainsbury S., Walter T., Nettleship J., Berrow N., Owens R., Gilbert R., Davidson A., Siddell S., Poon LL., Diprose J., Alderton D., Walsh M., Grimes JM., Stuart DI. Coronavirus - Interactions investigated in the context of Coronavirus dave@strubi.ox.ac.uk Only protein-protein interactions imex curation in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa predetermined predetermined participant predetermined Sutton G., Fry E., Carter L., Sainsbury S., Walter T., Nettleship J., Berrow N., Owens R., Gilbert R., Davidson A., Siddell S., Poon LL., Diprose J., Alderton D., Walsh M., Grimes JM., Stuart DI. Coronavirus - Interactions investigated in the context of Coronavirus SYBR Green II RNA gel stain is a sensitive nucleic acid gel stain that has bright fluorescence when bound to RNA. dave@strubi.ox.ac.uk Accepted 2020-MAY-12 AT 08:15 BST AT 08:15 BST by LIVIAP Structure (London, England : 1993) 2004 Only protein-protein interactions imex curation imex curation The nsp9 replicase protein of SARS-coronavirus, structure and functional insights. The nsp9 replicase protein of SARS-coronavirus, structure and functional insights. Structure (London, England : 1993) 2004 imex curation Sutton G., Fry E., Carter L., Sainsbury S., Walter T., Nettleship J., Berrow N., Owens R., Gilbert R., Davidson A., Siddell S., Poon LL., Diprose J., Alderton D., Walsh M., Grimes JM., Stuart DI. Coronavirus - Interactions investigated in the context of Coronavirus dave@strubi.ox.ac.uk Only protein-protein interactions imex curation in vitro In vitro dls dynamic light scattering dls predetermined predetermined participant predetermined Sutton G., Fry E., Carter L., Sainsbury S., Walter T., Nettleship J., Berrow N., Owens R., Gilbert R., Davidson A., Siddell S., Poon LL., Diprose J., Alderton D., Walsh M., Grimes JM., Stuart DI. Coronavirus - Interactions investigated in the context of Coronavirus dave@strubi.ox.ac.uk Accepted 2020-MAY-12 AT 08:15 BST AT 08:15 BST by LIVIAP Structure (London, England : 1993) 2004 Only protein-protein interactions imex curation imex curation The nsp9 replicase protein of SARS-coronavirus, structure and functional insights. The nsp9 replicase protein of SARS-coronavirus, structure and functional insights. Structure (London, England : 1993) 2004 imex curation Sutton G., Fry E., Carter L., Sainsbury S., Walter T., Nettleship J., Berrow N., Owens R., Gilbert R., Davidson A., Siddell S., Poon LL., Diprose J., Alderton D., Walsh M., Grimes JM., Stuart DI. Coronavirus - Interactions investigated in the context of Coronavirus dave@strubi.ox.ac.uk Only protein-protein interactions imex curation in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Sutton G., Fry E., Carter L., Sainsbury S., Walter T., Nettleship J., Berrow N., Owens R., Gilbert R., Davidson A., Siddell S., Poon LL., Diprose J., Alderton D., Walsh M., Grimes JM., Stuart DI. Coronavirus - Interactions investigated in the context of Coronavirus dave@strubi.ox.ac.uk Accepted 2020-MAY-12 AT 08:15 BST AT 08:15 BST by LIVIAP Structure (London, England : 1993) 2004 Only protein-protein interactions imex curation imex curation The nsp9 replicase protein of SARS-coronavirus, structure and functional insights. The nsp9 replicase protein of SARS-coronavirus, structure and functional insights. Structure (London, England : 1993) 2004 imex curation Sutton G., Fry E., Carter L., Sainsbury S., Walter T., Nettleship J., Berrow N., Owens R., Gilbert R., Davidson A., Siddell S., Poon LL., Diprose J., Alderton D., Walsh M., Grimes JM., Stuart DI. Coronavirus - Interactions investigated in the context of Coronavirus dave@strubi.ox.ac.uk Only protein-protein interactions imex curation in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Sutton G., Fry E., Carter L., Sainsbury S., Walter T., Nettleship J., Berrow N., Owens R., Gilbert R., Davidson A., Siddell S., Poon LL., Diprose J., Alderton D., Walsh M., Grimes JM., Stuart DI. Coronavirus - Interactions investigated in the context of Coronavirus dave@strubi.ox.ac.uk Accepted 2020-MAY-12 AT 08:15 BST AT 08:15 BST by LIVIAP Structure (London, England : 1993) 2004 Only protein-protein interactions imex curation imex curation p0c6x7-pro_0000037315 Non-structural protein 8 ORF1ab polyprotein 1a-1b rep protein protein sars-cov Human SARS coronavirus SARS-CoV Severe acute respiratory syndrome coronavirus AIASEFSSLPSYAAYATAQEAYEQAVANGDSEVVLKKLKKSLNVAKSEFDRDAAMQRKLEKMADQAMTQMYKQARSEDKRAKVTSAMQTMLFTMLRKLDNDALNNIINNARDGCVPLNIIPLTTAAKLMVVVPDYGTYKNTCDGNTFTYASALWEIQQVVDADSKIVQLSEINMDNSPNLAWPLIVTALRANSAVKLQ 3920 4117 E5D5A7840B20D23A unknown_ribonucleotide_oligo rna ribonucleic acid RNA rna chemical synthesis Chemical synthesis (Chemical synthesis) CGACUCAUGGACCUUGGCAG p0c6x7-pro_0000037316 Non-structural protein 9 rep ORF1ab polyprotein 1a-1b protein protein sars-cov Human SARS coronavirus SARS-CoV Severe acute respiratory syndrome coronavirus NNELSPVALRQMSCAAGTTQTACTDDNALAYYNNSKGGRFVLALLSDHQDLKWARFPKSDGTGTIYTELEPPCRFVTDTPKGPKVKYLYFIKGLNNLNRGMVLGSLAATVRLQ 4118 4230 EC021D1E6D6297A0 rep-81 3 7 unspecified role unspecified role neutral component neutral component purified purified n-terminus 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 rhinovirus 3C protease cleavage sequence tag 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 Table 4 rep-82 2 7 unspecified role unspecified role neutral component neutral component purified purified n-terminus 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 rhinovirus 3C protease cleavage sequence tag 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 Figure 4A It is not clear from the text if the N-terminal tags have been removed for DLS measurements, or not. For concentrations above 1.5mg/ml the Stokes’radius of approximately 2.1 nm (Figure 4A), is in close agreement with the calculated radius of 1.9 nm for a dimer. rep-83 4 7 unspecified role unspecified role neutral component neutral component purified purified n-terminus 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 rhinovirus 3C protease cleavage sequence tag 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 Figure 4B For tagged protein; at high concentration and high speed, nsp9 behaves as a dimer with an approximate Kd of 0.46mM. rep-84 1 6 unspecified role unspecified role bait bait chemical synthesis Chemical synthesis (Chemical synthesis) 7 unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 4D the oligo does not match to any sequence in NCBI that I can find -best match is NC_003714.1 residues 1-17 only (Pseudomonas phage phi-6 segment S). rep-85 4 5 unspecified role unspecified role neutral component neutral component purified purified n-terminus 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 rhinovirus 3C protease cleavage sequence tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component purified purified n-terminus 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 rhinovirus 3C protease cleavage sequence tag 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 Figure 4C rep-86 4 7 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 4B For untagged protein; nsp9 behaves as a dimer with an approximate Kd of 0.16mM which is not significantly different from the value obtained for the tagged protein.