MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Severe acute respiratory syndrome coronavirus ORF6 antagonizes STAT1 function by sequestering nuclear import factors on the rough endoplasmic reticulum/Golgi membrane. Severe acute respiratory syndrome coronavirus ORF6 antagonizes STAT1 function by sequestering nuclear import factors on the rough endoplasmic reticulum/Golgi membrane. Journal of virology 2007 imex curation Frieman M., Yount B., Heise M., Kopecky-Bromberg SA., Palese P., Baric RS. Coronavirus - Interactions investigated in the context of Coronavirus ude.cnu.liame@cirabr 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Frieman M., Yount B., Heise M., Kopecky-Bromberg SA., Palese P., Baric RS. Coronavirus - Interactions investigated in the context of Coronavirus ude.cnu.liame@cirabr Accepted 2020-APR-20 AT 15:04 BST AT 15:04 BST by LUANA Journal of virology 2007 Only protein-protein interactions imex curation imex curation Severe acute respiratory syndrome coronavirus ORF6 antagonizes STAT1 function by sequestering nuclear import factors on the rough endoplasmic reticulum/Golgi membrane. Severe acute respiratory syndrome coronavirus ORF6 antagonizes STAT1 function by sequestering nuclear import factors on the rough endoplasmic reticulum/Golgi membrane. Journal of virology 2007 imex curation Frieman M., Yount B., Heise M., Kopecky-Bromberg SA., Palese P., Baric RS. Coronavirus - Interactions investigated in the context of Coronavirus ude.cnu.liame@cirabr 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation chlae-vero Chlorocebus (Cercopithecus) aethiops kidney cells Grivet vero Chlorocebus (Cercopithecus) aethiops kidney cells similar to cos-7 cells but these are not trasnformed bifc bimolecular fluorescence complementation bifc tag fluorescence tag visualisation by fluorescence tag fluorescence Frieman M., Yount B., Heise M., Kopecky-Bromberg SA., Palese P., Baric RS. Coronavirus - Interactions investigated in the context of Coronavirus ude.cnu.liame@cirabr Accepted 2020-APR-20 AT 15:04 BST AT 15:04 BST by LUANA Journal of virology 2007 Only protein-protein interactions imex curation imex curation Severe acute respiratory syndrome coronavirus ORF6 antagonizes STAT1 function by sequestering nuclear import factors on the rough endoplasmic reticulum/Golgi membrane. Severe acute respiratory syndrome coronavirus ORF6 antagonizes STAT1 function by sequestering nuclear import factors on the rough endoplasmic reticulum/Golgi membrane. Journal of virology 2007 imex curation Frieman M., Yount B., Heise M., Kopecky-Bromberg SA., Palese P., Baric RS. Coronavirus - Interactions investigated in the context of Coronavirus ude.cnu.liame@cirabr 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation chlae-vero Chlorocebus (Cercopithecus) aethiops kidney cells Grivet vero Chlorocebus (Cercopithecus) aethiops kidney cells similar to cos-7 cells but these are not trasnformed fluorescence imaging fluorescence microscopy fluorescence imaging tag visualisation tag visualisation Frieman M., Yount B., Heise M., Kopecky-Bromberg SA., Palese P., Baric RS. Coronavirus - Interactions investigated in the context of Coronavirus ude.cnu.liame@cirabr Accepted 2020-APR-20 AT 15:04 BST AT 15:04 BST by LUANA Journal of virology 2007 Only protein-protein interactions imex curation imex curation a0a3g5biz0_sars Accessory protein 6 protein protein SARS coronavirus Urbani SARS coronavirus Urbani MFHLVDFQVTIAEILIIIMRTFRIAIWNLDVIISSIVRQLFKPLTKKNYSELDDEEPMELDYP 70C9150941C759BF stat2_human Signal transducer and activator of transcription 2 p113 STAT2 protein protein human Homo sapiens Human MAQWEMLQNLDSPFQDQLHQLYSHSLLPVDIRQYLAVWIEDQNWQEAALGSDDSKATMLFFHFLDQLNYECGRCSQDPESLLLQHNLRKFCRDIQPFSQDPTQLAEMIFNLLLEEKRILIQAQRAQLEQGEPVLETPVESQQHEIESRILDLRAMMEKLVKSISQLKDQQDVFCFRYKIQAKGKTPSLDPHQTKEQKILQETLNELDKRRKEVLDASKALLGRLTTLIELLLPKLEEWKAQQQKACIRAPIDHGLEQLETWFTAGAKLLFHLRQLLKELKGLSCLVSYQDDPLTKGVDLRNAQVTELLQRLLHRAFVVETQPCMPQTPHRPLILKTGSKFTVRTRLLVRLQEGNESLTVEVSIDRNPPQLQGFRKFNILTSNQKTLTPEKGQSQGLIWDFGYLTLVEQRSGGSGKGSNKGPLGVTEELHIISFTVKYTYQGLKQELKTDTLPVVIISNMNQLSIAWASVLWFNLLSPNLQNQQFFSNPPKAPWSLLGPALSWQFSSYVGRGLNSDQLSMLRNKLFGQNCRTEDPLLSWADFTKRESPPGKLPFWTWLDKILELVHDHLKDLWNDGRIMGFVSRSQERRLLKKTMSGTFLLRFSESSEGGITCSWVEHQDDDKVLIYSVQPYTKEVLQSLPLTEIIRHYQLLTEENIPENPLRFLYPRIPRDEAFGCYYQEKVNLQERRKYLKHRLIVVSNRQVDELQQPLELKPEPELESLELELGLVPEPELSLDLEPLLKAGLDLGPELESVLESTLEPVIEPTLCMVSQTVPEPDQGPVSQPVPEPDLPCDLRHLNTEPMEIFRNCVKIEEIMPNGDPLLAGQNTVDEVYVSRPSHFYTDGPLMPSDF DIP protein P52630 original sequence version: 116 E4C74674CB7A3215 ima5_human Importin subunit alpha-5 KPNA1 RCH2 Karyopherin subunit alpha-1 SRP1-beta RAG cohort protein 2 Nucleoprotein interactor 1 protein protein human Homo sapiens Human MTTPGKENFRLKSYKNKSLNPDEMRRRREEEGLQLRKQKREEQLFKRRNVATAEEETEEEVMSDGGFHEAQISNMEMAPGGVITSDMIEMIFSKSPEQQLSATQKFRKLLSKEPNPPIDEVISTPGVVARFVEFLKRKENCTLQFESAWVLTNIASGNSLQTRIVIQAGAVPIFIELLSSEFEDVQEQAVWALGNIAGDSTMCRDYVLDCNILPPLLQLFSKQNRLTMTRNAVWALSNLCRGKSPPPEFAKVSPCLNVLSWLLFVSDTDVLADACWALSYLSDGPNDKIQAVIDAGVCRRLVELLMHNDYKVVSPALRAVGNIVTGDDIQTQVILNCSALQSLLHLLSSPKESIKKEACWTISNITAGNRAQIQTVIDANIFPALISILQTAEFRTRKEAAWAITNATSGGSAEQIKYLVELGCIKPLCDLLTVMDSKIVQVALNGLENILRLGEQEAKRNGTGINPYCALIEEAYGLDKIEFLQSHENQEIYQKAFDLIEHYFGTEDEDSSIAPQVDLNQQQYIFQQCEAPMEGFQL DIP protein P52294 original sequence version: 72 DIP protein P52294 original sequence version: 90 DIP protein P52294 original sequence version: 112 DIP protein P52294 original sequence version: 160 E8407A3352D6051C stat1_human Signal transducer and activator of transcription 1-alpha/beta STAT1 Transcription factor ISGF-3 components p91/p84 protein protein human Homo sapiens Human MSQWYELQQLDSKFLEQVHQLYDDSFPMEIRQYLAQWLEKQDWEHAANDVSFATIRFHDLLSQLDDQYSRFSLENNFLLQHNIRKSKRNLQDNFQEDPIQMSMIIYSCLKEERKILENAQRFNQAQSGNIQSTVMLDKQKELDSKVRNVKDKVMCIEHEIKSLEDLQDEYDFKCKTLQNREHETNGVAKSDQKQEQLLLKKMYLMLDNKRKEVVHKIIELLNVTELTQNALINDELVEWKRRQQSACIGGPPNACLDQLQNWFTIVAESLQQVRQQLKKLEELEQKYTYEHDPITKNKQVLWDRTFSLFQQLIQSSFVVERQPCMPTHPQRPLVLKTGVQFTVKLRLLVKLQELNYNLKVKVLFDKDVNERNTVKGFRKFNILGTHTKVMNMEESTNGSLAAEFRHLQLKEQKNAGTRTNEGPLIVTEELHSLSFETQLCQPGLVIDLETTSLPVVVISNVSQLPSGWASILWYNMLVAEPRNLSFFLTPPCARWAQLSEVLSWQFSSVTKRGLNVDQLNMLGEKLLGPNASPDGLIPWTRFCKENINDKNFPFWLWIESILELIKKHLLPLWNDGCIMGFISKERERALLKDQQPGTFLLRFSESSREGAITFTWVERSQNGGEPDFHAVEPYTKKELSAVTFPDIIRNYKVMAAENIPENPLKYLYPNIDKDHAFGKYYSRPKEAPEPMELDGPKGTGYIKTELISVSEVHPSRLQTTDNLLPMSPEEFDEVSRIVGSVEFDSMMNTV 054A813522364BA6 imb1_human Importin subunit beta-1 Karyopherin subunit beta-1 Nuclear factor p97 Importin-90 KPNB1 NTF97 Pore targeting complex 97 kDa subunit protein protein human Homo sapiens Human MELITILEKTVSPDRLELEAAQKFLERAAVENLPTFLVELSRVLANPGNSQVARVAAGLQIKNSLTSKDPDIKAQYQQRWLAIDANARREVKNYVLQTLGTETYRPSSASQCVAGIACAEIPVNQWPELIPQLVANVTNPNSTEHMKESTLEAIGYICQDIDPEQLQDKSNEILTAIIQGMRKEEPSNNVKLAATNALLNSLEFTKANFDKESERHFIMQVVCEATQCPDTRVRVAALQNLVKIMSLYYQYMETYMGPALFAITIEAMKSDIDEVALQGIEFWSNVCDEEMDLAIEASEAAEQGRPPEHTSKFYAKGALQYLVPILTQTLTKQDENDDDDDWNPCKAAGVCLMLLATCCEDDIVPHVLPFIKEHIKNPDWRYRDAAVMAFGCILEGPEPSQLKPLVIQAMPTLIELMKDPSVVVRDTAAWTVGRICELLPEAAINDVYLAPLLQCLIEGLSAEPRVASNVCWAFSSLAEAAYEAADVADDQEEPATYCLSSSFELIVQKLLETTDRPDGHQNNLRSSAYESLMEIVKNSAKDCYPAVQKTTLVIMERLQQVLQMESHIQSTSDRIQFNDLQSLLCATLQNVLRKVQHQDALQISDVVMASLLRMFQSTAGSGGVQEDALMAVSTLVEVLGGEFLKYMEAFKPFLGIGLKNYAEYQVCLAAVGLVGDLCRALQSNIIPFCDEVMQLLLENLGNENVHRSVKPQILSVFGDIALAIGGEFKKYLEVVLNTLQQASQAQVDKSDYDMVDYLNELRESCLEAYTGIVQGLKGDQENVHPDVMLVQPRVEFILSFIDHIAGDEDHTDGVVACAAGLIGDLCTAFGKDVLKLVEARPMIHELLTEGRRSKTNKAKTLATWATKELRKLKNQA F3BB8B73E7E51639 ima1_human Importin subunit alpha-1 KPNA2 RCH1 SRP1 Karyopherin subunit alpha-2 SRP1-alpha RAG cohort protein 1 protein protein human Homo sapiens Human MSTNENANTPAARLHRFKNKGKDSTEMRRRRIEVNVELRKAKKDDQMLKRRNVSSFPDDATSPLQENRNNQGTVNWSVDDIVKGINSSNVENQLQATQAARKLLSREKQPPIDNIIRAGLIPKFVSFLGRTDCSPIQFESAWALTNIASGTSEQTKAVVDGGAIPAFISLLASPHAHISEQAVWALGNIAGDGSVFRDLVIKYGAVDPLLALLAVPDMSSLACGYLRNLTWTLSNLCRNKNPAPPIDAVEQILPTLVRLLHHDDPEVLADTCWAISYLTDGPNERIGMVVKTGVVPQLVKLLGASELPIVTPALRAIGNIVTGTDEQTQVVIDAGALAVFPSLLTNPKTNIQKEATWTMSNITAGRQDQIQQVVNHGLVPFLVSVLSKADFKTQKEAVWAVTNYTSGGTVEQIVYLVHCGIIEPLMNLLTAKDTKIILVILDAISNIFQAAEKLGETEKLSIMIEECGGLDKIEALQNHENESVYKASLSLIEKYFSVEEEEDQNVVPETTSEGYTFQVQDGAPGTFNF DIP protein P52292 original sequence version: 180 DIP protein P52292 original sequence version: 128 B0F94A0475B80EED kpna1-stat1-1 1 5 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 6 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association fig 3a IFN-beta kpna2-a0a3g5biz0_cvhsa-1 1 8 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association fig. 3b 1-139 3 9 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization fig. 3d a0a3g5biz0_cvhsa-kpna2-1 2 9 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed living cell living cell nYFP yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed living cell living cell cYFP yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association fig. 4 a-e 1-140 3 4 anti tag immunost anti tag immunostaining anti tag immunost tag visualisation tag visualisation unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 anti tag immunost anti tag immunostaining anti tag immunost tag visualisation tag visualisation unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization fig. 5a kpna2-a0a3g5biz0_cvhsa-2 1 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed Flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association fig. 5b 1-141 1 6 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 8 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level association association fig. 6a lane 1-2 INF-beta kpna1-kpnb1-3 1 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed binding region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed physical association physical association fig. 6a lane 3-6 INF-beta kpna1-kpnb1-4 1 6 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed Flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association fig. 6a lane 9-10 INF-beta As shown in Fig.3A, KPNA1 binds to STAT1 only after treatment with IFN-β (Fig.6A, lane 2). Consistent with previous findings, this interaction is independent of ORF6 expression (Fig.6A, lanes 9 and 10), since in the presence of ORF6, KPNA1 still binds STAT1 after IFN-β treatment. kpna2-kpnb1-7 1 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed binding region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain Flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association fig. 6a lane 11-12 INF-beta kpna2-kpnb1-8 1 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed binding region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain Flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed association association fig. 6a lane 11-14 INF-beta Moreover, Western blotting of the Flag immunoprecipitates for KPNB1 shows that in the absence of ORF6, KPNB1 associates with KPNA1 and KPNA2 but not KPNA2ΔN, confirming that the importin beta binding site of KPNA2 is necessary for KPNB1 binding. In the presence of ORF6, KPNA1 no longer binds KPNB1; however, under identical conditions, KPNA2 binds KPNB1 efficiently (Fig.6A, lanes 9 and 11). As predicted, KPNA2 binds ORF6 but no longer interacts with KPNB1 (Fig.6A, lane 13). Together, these results indicate that ORF6 binds KPNA2 as well as KPNB1; however, this binding does not reduce the KPNA1:STAT1 complex in the cell, and it reduces only the KPNA1:STAT1:KPNB1 complex. a0a3g5biz0_cvhsa-kpna2-2 3 9 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell A0A3G5BIZ0:p.Asp54_Met58delinsAlaAlaAlaAlaAla p.asp54_met58delinsalaalaalaalaala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain DEEPM AAAAA Tue Jun 30 18:12:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp54_Met58delinsAlaAlaAlaAlaAla' ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined A0A3G5BIZ0:p.Glu59_Pro63delinsAlaAlaAlaAlaAla p.glu59_pro63delinsalaalaalaalaala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain ELDYP AAAAA Tue Jun 30 18:00:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu59_Pro63delinsAlaAlaAlaAlaAla' colocalization colocalization fig. 8 We further characterized the alanine mutants by determining whether KPNA2 would be retained in the ER/Golgi membrane. Under conditions in which ORF654-58Ala and ORF659-63Ala did not retain KPNA2, the ORF649-53Ala product still retained KPNA2 at the ER/Golgi membrane by confocal microscopy (Fig.8).