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/ SARS-CoV-2 proteases PLpro and 3CLpro cleave IRF3 and critical modulators of inflammatory pathways (NLRP12 and TAB1): implications for disease presentation across species. SARS-CoV-2 proteases PLpro and 3CLpro cleave IRF3 and critical modulators of inflammatory pathways (NLRP12 and TAB1): implications for disease presentation across species. Emerging microbes & infections 2020 imex curation Moustaqil M., Ollivier E., Chiu HP., Van Tol S., Rudolffi-Soto P., Stevens C., Bhumkar A., Hunter DJB., Freiberg AN., Jacques D., Lee B., Sierecki E., Gambin Y. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus y.gambin@unsw.edu.au,e.sierecki@unsw.edu.au,benhur.lee@mssm.edu 29-Mar-2021: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro protease assay protease assay tag fluorescence tag visualisation by fluorescence tag fluorescence Moustaqil M., Ollivier E., Chiu HP., Van Tol S., Rudolffi-Soto P., Stevens C., Bhumkar A., Hunter DJB., Freiberg AN., Jacques D., Lee B., Sierecki E., Gambin Y. Virus - Publications including interactions involving viral proteins I do not regard the M.wt of the fragments produced after cleavage, sufficient to enter a cleavage site(s), (as a ptm feature(s)). However I cannot access Supp. figures after figure S6, but I do not think they show any additional experimental evidence. Although this is an enzymatic assay the substrates are not purified (although gels do not look unclean) so I cannot enter Interaction Type as any child of a "direct" interaction. However I do not know what else to call it, other than some sort of Protease assay? So maybe they should be entered as "protein cleavage" for Interaction type? Samples were loaded onto SDS-page gels; the proteins were detected by scanning the gel for green (GFP) or red (mCherry) fluorescence and proteolytic cleavage was assessed from the changes in banding patterns. y.gambin@unsw.edu.au,e.sierecki@unsw.edu.au,benhur.lee@mssm.edu Accepted 2021-JAN-26 AT 08:14 GMT AT 08:14 GMT by ORCHARD Emerging microbes & infections 2020 Only protein-protein interactions imex curation imex curation nal12_mouse NACHT, LRR and PYD domains-containing protein 12 Monarch-1 PYRIN-containing APAF1-like protein 7 Nlrp12 NALP12 protein protein mouse Mus musculus Mouse MLPSTARDGLYRLSTYLEELEAGELKKFKLFLGIAEDLSQDKIPWGRMEKAGPLEMAQLMVAHMGTREAWLLALSTFQRIHRKDLWERGQGEDLVRVTPNNGLCLFESQSACPLDVSPNAPRKDLQTTYKDYVRRKFQLMEDRNARLGECVNLSNRYTRLLLVKEHSNPIWTQQKFVDVEWERSRTRRHQTSPIQMETLFEPDEERPEPPHTVVLQGAAGMGKSMLAHKVMLDWADGRLFQGRFDYVFYISCRELNRSHTQCSVQDLISSCWPERGISLEDLMQAPDRLLFIIDGFDKLHPSFHDAQGPWCLCWEEKQPTEVLLGSLIRRLLLPQVSLLITTRPCALEKLHGLLEHPRHVEILGFSEEARKEYFYRYFHNTGQASRVLSFLMDYEPLFTMCFVPMVSWVVCTCLKQQLESGELLRQTPRTTTAVYMFYLLSLMQPKPGTPTFKVPANQRGLVSLAAEGLWNQKILFDEQDLGKHGLDGADVSTFLNVNIFQKGIKCEKFYSFIHLSFQEFFAAMYCALNGREAVRRALAEYGFSERNFLALTVHFLFGLLNEEMRCYLERNLGWSISPQVKEEVLAWIQNKAGSEGSTLQHGSLELLSCLYEVQEEDFIQQALSHFQVVVVRSISTKMEHMVCSFCARYCRSTEVLHLHGSAYSTGMEDDPPEPSGVQTQSTYLQERNMLPDVYSAYLSAAVCTNSNLIELALYRNALGSQGVRLLCQGLRHASCKLQNLRLKRCQISGSACQDLAAAVIANRNLIRLDLSDNSIGVPGLELLCEGLQHPRCRLQMIQLRKCLLEAAAGRSLASVLSNNSYLVELDLTGNPLEDSGLKLLCQGLRHPVCRLRTLWLKICHLGQASCEDLASTLKMNQSLLELDLGLNDLGDSGVLLLCEGLSHPDCKLQTLRLGICRLGSVACVGIASVLQVNTCLQELDLSFNDLGDRGLQLLGEGLRHQTCRLQKLWLDNCGLTSKACEDLSSILGISQTLHELYLTNNALGDTGVCLLCKRLRHPGCKLRVLWLFGMDLNKKTHRRMAALRVTKPYLDIGC 3F17BEA78BA38621 p0dtd1-pro_0000449623 3C-like proteinase nsp5 rep ORF1ab polyprotein 1a-1b protein protein SARS-CoV-2 SARS-CoV-2 SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 2019-nCoV SGFRKMAFPSGKVEGCMVQVTCGTTTLNGLWLDDVVYCPRHVICTSEDMLNPNYEDLLIRKSNHNFLVQAGNVQLRVIGHSMQNCVLKLKVDTANPKTPKYKFVRIQPGQTFSVLACYNGSPSGVYQCAMRPNFTIKGSFLNGSCGSVGFNIDYDCVSFCYMHHMELPTGVHAGTDLEGNFYGPFVDRQTAQAAGTDTTITVNVLAWLYAAVINGDRWFLNRFTTTLNDFNLVAMKYNYEPLTQDHVDILGPLSAQTGIAVLDMCASLKELLQNGMNGRTILGSALLEDEFTPFDVVRQCSGVTFQ 3264 3569 p0dtd1-pro_0000449621 Papain-like protease nsp3 rep ORF1ab polyprotein 1a-1b protein protein SARS-CoV-2 SARS-CoV-2 SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 2019-nCoV APTKVTFGDDTVIEVQGYKSVNITFELDERIDKVLNEKCSAYTVELGTEVNEFACVVADAVIKTLQPVSELLTPLGIDLDEWSMATYYLFDESGEFKLASHMYCSFYPPDEDEEEGDCEEEEFEPSTQYEYGTEDDYQGKPLEFGATSAALQPEEEQEEDWLDDDSQQTVGQQDGSEDNQTTTIQTIVEVQPQLEMELTPVVQTIEVNSFSGYLKLTDNVYIKNADIVEEAKKVKPTVVVNAANVYLKHGGGVAGALNKATNNAMQVESDDYIATNGPLKVGGSCVLSGHNLAKHCLHVVGPNVNKGEDIQLLKSAYENFNQHEVLLAPLLSAGIFGADPIHSLRVCVDTVRTNVYLAVFDKNLYDKLVSSFLEMKSEKQVEQKIAEIPKEEVKPFITESKPSVEQRKQDDKKIKACVEEVTTTLEETKFLTENLLLYIDINGNLHPDSATLVSDIDITFLKKDAPYIVGDVVQEGVLTAVVIPTKKAGGTTEMLAKALRKVPTDNYITTYPGQGLNGYTVEEAKTVLKKCKSAFYILPSIISNEKQEILGTVSWNLREMLAHAEETRKLMPVCVETKAIVSTIQRKYKGIKIQEGVVDYGARFYFYTSKTTVASLINTLNDLNETLVTMPLGYVTHGLNLEEAARYMRSLKVPATVSVSSPDAVTAYNGYLTSSSKTPEEHFIETISLAGSYKDWSYSGQSTQLGIEFLKRGDKSVYYTSNPTTFHLDGEVITFDNLKTLLSLREVRTIKVFTTVDNINLHTQVVDMSMTYGQQFGPTYLDGADVTKIKPHNSHEGKTFYVLPNDDTLRVEAFEYYHTTDPSFLGRYMSALNHTKKWKYPQVNGLTSIKWADNNCYLATALLTLQQIELKFNPPALQDAYYRARAGEAANFCALILAYCNKTVGELGDVRETMSYLFQHANLDSCKRVLNVVCKTCGQQQTTLKGVEAVMYMGTLSYEQFKKGVQIPCTCGKQATKYLVQQESPFVMMSAPPAQYELKHGTFTCASEYTGNYQCGHYKHITSKETLYCIDGALLTKSSEYKGPITDVFYKENSYTTTIKPVTYKLDGVVCTEIDPKLDNYYKKDNSYFTEQPIDLVPNQPYPNASFDNFKFVCDNIKFADDLNQLTGYKKPASRELKVTFFPDLNGDVVAIDYKHYTPSFKKGAKLLHKPIVWHVNNATNKATYKPNTWCIRCLWSTKPVETSNSFDVLKSEDAQGMDNLACEDLKPVSEEVVENPTIQKDVLECNVKTTEVVGDIILKPANNSLKITEEVGHTDLMAAYVDNSSLTIKKPNELSRVLGLKTLATHGLAAVNSVPWDTIANYAKPFLNKVVSTTTNIVTRCLNRVCTNYMPYFFTLLLQLCTFTRSTNSRIKASMPTTIAKNTVKSVGKFCLEASFNYLKSPNFSKLINIIIWFLLLSVCLGSLIYSTAALGVLMSNLGMPSYCTGYREGYLNSTNVTIATYCTGSIPCSVCLSGLDSLDTYPSLETIQITISSFKWDLTAFGLVAEWFLAYILFTRFFYVLGLAAIMQLFFSYFAVHFISNSWLMWLIINLVQMAPISAMVRMYIFFASFYYVWKSYVHVVDGCNSSTCMMCYKRNRATRVECTTIVNGVRRSFYVYANGGKGFCKLHNWNCVNCDTFCAGSTFISDEVARDLSLQFKRPINPTDQSSYIVDSVTVKNGSIHLYFDKAGQKTYERHSLSHFVNLDNLRANNTKGSLPINVIVFDGKSKCEESSAKSASVYYSQLMCQPILLLDQALVSDVGDSAEVAVKMFDAYVNTFSSTFNVPMEKLKTLVATAEAELAKNVSLDNVLSTFISAARQGFVDSDVETKDVVECLKLSHQSDIEVTGDSCNNYMLTYNKVENMTPRDLGACIDCSARHINAQVAKSHNIALIWNVKDFMSLSEQLRKQIRSAAKKNNLPFKLTCATTRQVVNVVTTKIALKGG 819 2763 tab1_human TGF-beta-activated kinase 1 and MAP3K7-binding protein 1 TAB1 MAP3K7IP1 TGF-beta-activated kinase 1-binding protein 1 Mitogen-activated protein kinase kinase kinase 7-interacting protein 1 protein protein human Homo sapiens Human MAAQRRSLLQSEQQPSWTDDLPLCHLSGVGSASNRSYSADGKGTESHPPEDSWLKFRSENNCFLYGVFNGYDGNRVTNFVAQRLSAELLLGQLNAEHAEADVRRVLLQAFDVVERSFLESIDDALAEKASLQSQLPEGVPQHQLPPQYQKILERLKTLEREISGGAMAVVAVLLNNKLYVANVGTNRALLCKSTVDGLQVTQLNVDHTTENEDELFRLSQLGLDAGKIKQVGIICGQESTRRIGDYKVKYGYTDIDLLSAAKSKPIIAEPEIHGAQPLDGVTGFLVLMSEGLYKALEAAHGPGQANQEIAAMIDTEFAKQTSLDAVAQAVVDRVKRIHSDTFASGGERARFCPRHEDMTLLVRNFGYPLGEMSQPTPSPAPAAGGRVYPVSVPYSSAQSTSKTSVTLSLVMPSQGQMVNGAHSASTLDEATPTLTNQSPTLTLQSTNTHTQSSSSSSDGGLFRSRPAHSLPPGEDGRVEPYVDFAEFYRLWSVDHGEQSVVTAP DIP protein Q15750 original sequence version: 74 DIP protein Q15750 original sequence version: 110 DIP protein Q15750 original sequence version: 111 A45743288718983A nal12_human NACHT, LRR and PYD domains-containing protein 12 NLRP12 NALP12 PYPAF7 RNO Monarch-1 PYRIN-containing APAF1-like protein 7 Regulated by nitric oxide protein protein human Homo sapiens Human MLRTAGRDGLCRLSTYLEELEAVELKKFKLYLGTATELGEGKIPWGSMEKAGPLEMAQLLITHFGPEEAWRLALSTFERINRKDLWERGQREDLVRDTPPGGPSSLGNQSTCLLEVSLVTPRKDPQETYRDYVRRKFRLMEDRNARLGECVNLSHRYTRLLLVKEHSNPMQVQQQLLDTGRGHARTVGHQASPIKIETLFEPDEERPEPPRTVVMQGAAGIGKSMLAHKVMLDWADGKLFQGRFDYLFYINCREMNQSATECSMQDLIFSCWPEPSAPLQELIRVPERLLFIIDGFDELKPSFHDPQGPWCLCWEEKRPTELLLNSLIRKKLLPELSLLITTRPTALEKLHRLLEHPRHVEILGFSEAERKEYFYKYFHNAEQAGQVFNYVRDNEPLFTMCFVPLVCWVVCTCLQQQLEGGGLLRQTSRTTTAVYMLYLLSLMQPKPGAPRLQPPPNQRGLCSLAADGLWNQKILFEEQDLRKHGLDGEDVSAFLNMNIFQKDINCERYYSFIHLSFQEFFAAMYYILDEGEGGAGPDQDVTRLLTEYAFSERSFLALTSRFLFGLLNEETRSHLEKSLCWKVSPHIKMDLLQWIQSKAQSDGSTLQQGSLEFFSCLYEIQEEEFIQQALSHFQVIVVSNIASKMEHMVSSFCLKRCRSAQVLHLYGATYSADGEDRARCSAGAHTLLVQLPERTVLLDAYSEHLAAALCTNPNLIELSLYRNALGSRGVKLLCQGLRHPNCKLQNLRLKRCRISSSACEDLSAALIANKNLTRMDLSGNGVGFPGMMLLCEGLRHPQCRLQMIQLRKCQLESGACQEMASVLGTNPHLVELDLTGNALEDLGLRLLCQGLRHPVCRLRTLWLKICRLTAAACDELASTLSVNQSLRELDLSLNELGDLGVLLLCEGLRHPTCKLQTLRLGICRLGSAACEGLSVVLQANHNLRELDLSFNDLGDWGLWLLAEGLQHPACRLQKLWLDSCGLTAKACENLYFTLGINQTLTDLYLTNNALGDTGVRLLCKRLSHPGCKLRVLWLFGMDLNKMTHSRLAALRVTKPYLDIGC 8C10AFE4907C131B irf3_human Interferon regulatory factor 3 IRF3 protein protein human Homo sapiens Human MGTPKPRILPWLVSQLDLGQLEGVAWVNKSRTRFRIPWKHGLRQDAQQEDFGIFQAWAEATGAYVPGRDKPDLPTWKRNFRSALNRKEGLRLAEDRSKDPHDPHKIYEFVNSGVGDFSQPDTSPDTNGGGSTSDTQEDILDELLGNMVLAPLPDPGPPSLAVAPEPCPQPLRSPSLDNPTPFPNLGPSENPLKRLLVPGEEWEFEVTAFYRGRQVFQQTISCPEGLRLVGSEVGDRTLPGWPVTLPDPGMSLTDRGVMSYVRHVLSCLGGGLALWRAGQWLWAQRLGHCHTYWAVSEELLPNSGHGPDGEVPKDKEGGVFDLGPFIVDLITFTEGSGRSPRYALWFCVGESWPQDQPWTKRLVMVKVVPTCLRALVEMARVGGASSLENTVDLHISNSHPLSLTSDQYKAYLQDLVEGMDFQGPGES DIP protein Q14653 original sequence version: 96 DIP protein Q14653 original sequence version: 75 DIP protein Q14653 original sequence version: 95 DIP protein Q14653-1 original sequence version: 128 DIP protein Q14653 original sequence version: 119 F536676FA78B0110 irf3-rep-7 1 7 enzyme target enzyme target substrate substrate neutral component neutral component cell lysate cell lysate n-terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range leita Leishmania tarentolae Sauroleishmania tarentolae 4 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) protein cleavage protein cleavage 1B,2, Supp. figure 3 The predicted cleavage site is after residue 271, after the sequence CLGGG. I do not regard the M.wt of the 2 fragments produced after cleavage, sufficient to enter aa271 as a cleavage site, (as a ptm feature). irf3-rep-8 1 5 enzyme target enzyme target substrate substrate neutral component neutral component cell lysate cell lysate n-terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range leita Leishmania tarentolae Sauroleishmania tarentolae 3 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) protein cleavage protein cleavage 3B The gel shows two additional bands upon cleavage. rep-tab1-1 1 5 enzyme target enzyme target substrate substrate neutral component neutral component cell lysate cell lysate C-terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range leita Leishmania tarentolae Sauroleishmania tarentolae 3 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) protein cleavage protein cleavage 3A The gel shows two additional bands upon cleavage. rep-tab1-2 1 6 enzyme target enzyme target substrate substrate neutral component neutral component cell lysate cell lysate C-terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range leita Leishmania tarentolae Sauroleishmania tarentolae 3 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) protein cleavage protein cleavage 4A The gel shows two additional bands upon cleavage. nlrp12-rep-1 1 6 enzyme target enzyme target substrate substrate neutral component neutral component cell lysate cell lysate C-terminus mcherry mcherry fluorescent protein tag mcherry c-term range C-terminal range c-term range c-term range C-terminal range c-term range leita Leishmania tarentolae Sauroleishmania tarentolae 3 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) protein cleavage protein cleavage 4E The gel shows two additional bands upon cleavage. nlrp12-rep-2 1 2 enzyme target enzyme target substrate substrate neutral component neutral component cell lysate cell lysate C-terminus mcherry mcherry fluorescent protein tag mcherry c-term range C-terminal range c-term range c-term range C-terminal range c-term range leita Leishmania tarentolae Sauroleishmania tarentolae 3 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) protein cleavage protein cleavage 4E mouse NLRP12 is only cleaved once. nlrp12-rep-3 1 3 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 6 enzyme target enzyme target substrate substrate neutral component neutral component cell lysate cell lysate n-terminus mcherry mcherry fluorescent protein tag mcherry n-term range n-terminal range n-term range n-term range n-terminal range n-term range leita Leishmania tarentolae Sauroleishmania tarentolae protein cleavage protein cleavage 4E The gel shows two additional bands upon cleavage. nlrp12-rep-4 1 2 enzyme target enzyme target substrate substrate neutral component neutral component cell lysate cell lysate n-terminus mcherry mcherry fluorescent protein tag mcherry n-term range n-terminal range n-term range n-term range n-terminal range n-term range leita Leishmania tarentolae Sauroleishmania tarentolae 3 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) protein cleavage protein cleavage 4E mouse NLRP12 is only cleaved once.