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/ http://www.ebi.ac.uk/ brandherm-2021-1 in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Brandherm L., Kobaš AM., Klöhn M., Brüggemann Y., Pfaender S., Rassow J., Kreimendahl S. Virus - Publications including interactions involving viral proteins joachim.rassow@rub.de International journal of molecular sciences 2021 Only protein-protein interactions imex curation imex curation Coronavirus - Interactions investigated in the context of Coronavirus brandherm-2021-2 ecoli Escherichia coli (strain K12) molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Brandherm L., Kobaš AM., Klöhn M., Brüggemann Y., Pfaender S., Rassow J., Kreimendahl S. Virus - Publications including interactions involving viral proteins joachim.rassow@rub.de International journal of molecular sciences 2021 Only protein-protein interactions imex curation imex curation Coronavirus - Interactions investigated in the context of Coronavirus brandherm-2021-3 in vitro In vitro enzymatic footprint enzymatic footprinting enzymatic footprint weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Brandherm L., Kobaš AM., Klöhn M., Brüggemann Y., Pfaender S., Rassow J., Kreimendahl S. Virus - Publications including interactions involving viral proteins joachim.rassow@rub.de International journal of molecular sciences 2021 Only protein-protein interactions imex curation imex curation Coronavirus - Interactions investigated in the context of Coronavirus brandherm-2021-4 rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes bn-page blue native page bn-page weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Brandherm L., Kobaš AM., Klöhn M., Brüggemann Y., Pfaender S., Rassow J., Kreimendahl S. Virus - Publications including interactions involving viral proteins joachim.rassow@rub.de International journal of molecular sciences 2021 Only protein-protein interactions imex curation imex curation Coronavirus - Interactions investigated in the context of Coronavirus orf9b_sars2 ORF9b protein Accessory protein 9b ORF-9b Protein 9b 9b protein protein SARS-CoV-2 SARS-CoV-2 SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 2019-nCoV MDPKISEMHPALRLVDPQIQLAVTRMENAVGRDQNNVGPKVYPIILRLGSPLSLNMARKTLNSLEDKAFQLTPIAVQMTKLATTEELPDEFVVVTVK tom70_human Mitochondrial import receptor subunit TOM70 TOMM70 KIAA0719 TOM70 TOMM70A Translocase of outer membrane 70 kDa subunit Translocase of outer mitochondrial membrane protein 70 Mitochondrial precursor proteins import receptor protein protein human Homo sapiens Human MAASKPVEAAVVAAAVPSSGSGVGGGGTAGPGTGGLPRWQLALAVGAPLLLGAGAIYLWSRQQRRREARGRGDASGLKRNSERKTPEGRASPAPGSGHPEGPGAHLDMNSLDRAQAAKNKGNKYFKAGKYEQAIQCYTEAISLCPTEKNVDLSTFYQNRAAAFEQLQKWKEVAQDCTKAVELNPKYVKALFRRAKAHEKLDNKKECLEDVTAVCILEGFQNQQSMLLADKVLKLLGKEKAKEKYKNREPLMPSPQFIKSYFSSFTDDIISQPMLKGEKSDEDKDKEGEALEVKENSGYLKAKQYMEEENYDKIISECSKEIDAEGKYMAEALLLRATFYLLIGNANAAKPDLDKVISLKEANVKLRANALIKRGSMYMQQQQPLLSTQDFNMAADIDPQNADVYHHRGQLKILLDQVEEAVADFDECIRLRPESALAQAQKCFALYRQAYTGNNSSQIQAAMKGFEEVIKKFPRCAEGYALYAQALTDQQQFGKADEMYDKCIDLEPDNATTYVHKGLLQLQWKQDLDRGLELISKAIEIDNKCDFAYETMGTIEVQRGNMEKAIDMFNKAINLAKSEMEMAHLYSLCDAAHAQTEVAKKYGLKPPTL hs90b_human Heat shock protein HSP 90-beta HSP90AB1 HSP90B HSPC2 HSPCB Heat shock 84 kDa protein protein human Homo sapiens Human MPEEVHHGEEEVETFAFQAEIAQLMSLIINTFYSNKEIFLRELISNASDALDKIRYESLTDPSKLDSGKELKIDIIPNPQERTLTLVDTGIGMTKADLINNLGTIAKSGTKAFMEALQAGADISMIGQFGVGFYSAYLVAEKVVVITKHNDDEQYAWESSAGGSFTVRADHGEPIGRGTKVILHLKEDQTEYLEERRVKEVVKKHSQFIGYPITLYLEKEREKEISDDEAEEEKGEKEEEDKDDEEKPKIEDVGSDEEDDSGKDKKKKTKKIKEKYIDQEELNKTKPIWTRNPDDITQEEYGEFYKSLTNDWEDHLAVKHFSVEGQLEFRALLFIPRRAPFDLFENKKKKNNIKLYVRRVFIMDSCDELIPEYLNFIRGVVDSEDLPLNISREMLQQSKILKVIRKNIVKKCLELFSELAEDKENYKKFYEAFSKNLKLGIHEDSTNRRRLSELLRYHTSQSGDEMTSLSEYVSRMKETQKSIYYITGESKEQVANSAFVERVRKRGFEVVYMTEPIDEYCVQQLKEFDGKSLVSVTKEGLELPEDEEEKKKMEESKAKFENLCKLMKEILDKKVEKVTISNRLVSSPCCIVTSTYGWTANMERIMKAQALRDNSTMGYMMAKKHLEINPDHPIVETLRQKAEADKNDKAVKDLVVLLFETALLSSGFSLEDPQTHSNRIYRMIKLGLGIDEDEVAAEEPNAAVPDEIPPLEGDEDASRMEEVD adt3_human ADP/ATP translocase 3 SLC25A6 ANT3 CDABP0051 Adenine nucleotide translocator 3 ADP,ATP carrier protein 3 Solute carrier family 25 member 6 ADP,ATP carrier protein, isoform T2 AAC3 protein protein human Homo sapiens Human MTEQAISFAKDFLAGGIAAAISKTAVAPIERVKLLLQVQHASKQIAADKQYKGIVDCIVRIPKEQGVLSFWRGNLANVIRYFPTQALNFAFKDKYKQIFLGGVDKHTQFWRYFAGNLASGGAAGATSLCFVYPLDFARTRLAADVGKSGTEREFRGLGDCLVKITKSDGIRGLYQGFSVSVQGIIIYRAAYFGVYDTAKGMLPDPKNTHIVVSWMIAQTVTAVAGVVSYPFDTVRRRMMMQSGRKGADIMYTGTVDCWRKIFRDEGGKAFFKGAWSNVLRGMGGAFVLVLYDELKKVI tomm70-orf9b_sars2-1 4086907 N/A 4086911 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated P0DTD2:p.Ser53Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P0DTD2:p.Ser53Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes N/A 4086912 unspecified role unspecified role bait bait purified purified O94826:p.Glu477Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false 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 false cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false 1C/I/K A mutation of either amino acid residue (TOM70-E477A OR Of9b-S53E) drastically reduces the association of the two proteins. The incubation of phosphomimetic Orf9bS53E with isolated TOM70E477A did not lead to a reduction in comparison to TOM70WT. tomm70-hsp90ab1-1 4086907 N/A 4086913 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes N/A 4086912 unspecified role unspecified role bait bait purified purified O94826:p.Glu477Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false 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 false cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false O94826:p.Arg192Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false Figs. 3 B/C, 4C Orf9b (P0DTD2), Wild type only. No effect was observed when TOM70 was pre-incubated with the phosphomimetic variant Orf9bS53E. tomm70-slc25a6-1 4086907 N/A 4086914 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes N/A 4086912 unspecified role unspecified role bait bait purified purified O94826:p.Glu477Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false 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 false cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 2D Orf9b (P0DTD2). tomm70-orf9b_sars2-2 4086907 N/A 4086911 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes N/A 4086912 unspecified role unspecified role bait bait purified purified O94826:p.Arg192Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false 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 false cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false 4E/F/G 400 mM KCl, but only for WT interaction. orf9b_sars2-24 4086908 N/A 4086911 unspecified role unspecified role neutral component neutral component purified purified P0DTD2:p.Ser53Glu mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 2.0 direct interaction direct interaction false false false 1F Both Orf9b variants were eluted as dimers of a molecular mass of approximately 28.7 kDa. tomm70-orf9b_sars2-3 4086909 N/A 4086911 unspecified role unspecified role neutral component neutral component in vitro translated in vitro translated protein in vitro translated Dihydrofolate reductase tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false P0DTD2:p.Ser53Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P0DTD2:p.Ser53Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes N/A 4086912 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 false cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false Supp. figure 2 Orf9bWT-DHFR is efficiently bound to the C-terminal domain of TOM70, with this part of the construct being shielded from trypsin degradation. The Orf9b S53A mutant showed the same fragmentation pattern as the WT, but Orf9b S53E showed much less protection in the presence of TOM70, indicating reduced binding of both proteins. orf9b_sars2-orf9b_sars2-1 4086910 N/A 4086911 unspecified role unspecified role neutral component neutral component in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 4086911 unspecified role unspecified role neutral component neutral component in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Supp. figure 3 Most of the Orf9b was found as a component of a high molecular weight complex of 220–240 kDa.