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/ wang-2005c-1 Domain interactions within the Ski2/3/8 complex and between the Ski complex and Ski7p. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Wang L., Lewis MS., Johnson AW. arlen@mail.utexas.edu RNA (1355-8382) 2005 wang-2005c-2 Domain interactions within the Ski2/3/8 complex and between the Ski complex and Ski7p. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Wang L., Lewis MS., Johnson AW. arlen@mail.utexas.edu RNA (1355-8382) 2005 wang-2005c-3 Domain interactions within the Ski2/3/8 complex and between the Ski complex and Ski7p. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast pull down pull down western blot western blot Immuno blot Immuno blot Wang L., Lewis MS., Johnson AW. arlen@mail.utexas.edu RNA (1355-8382) 2005 wang-2005c-4 Domain interactions within the Ski2/3/8 complex and between the Ski complex and Ski7p. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Wang L., Lewis MS., Johnson AW. arlen@mail.utexas.edu RNA (1355-8382) 2005 wang-2005c-5 Domain interactions within the Ski2/3/8 complex and between the Ski complex and Ski7p. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast density sedimentation cosedimentation through density gradient density sedimentation western blot western blot Immuno blot Immuno blot Wang L., Lewis MS., Johnson AW. Complex partialy purified by GST-pulldown then by 10-20% glycerol gradient centrifugation arlen@mail.utexas.edu RNA (1355-8382) 2005 ski3_yeast Superkiller protein 3 SKI3 P9677.7 YPR189W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSDIKQLLKEAKQELTNRDYEETIEISEKVLKLDPDNYFAHIFLGKALSSLPASNNVSSNRNLERATNHYVSAAKLVPDNLLAWKGLFLLFRTTEVVPDILSYDEYFDLCGQYADALLKQEQSQVELINDIKLLKKTHPDCQKAFYQHLKPGSLMAETIGRHLSTPQDALLNLIKILSNIETTEIGKTLSQNRLKLKASDPDYQIKLNSFSWEIIKNSEIDQLYNQLVNILADDQKRSEIENQWLEYRIKVLKSMPLDVKKDFFTKVKEMVEDMVLVNHQSLLAWQKYFEWTDYEDLDNMDAPLIIKYFKKFPKDPLAMILYSWLSSKLSKYDIKSLESANKPPEGHKKTEKETDIKDVDETNEDEVKDRVEDEVKDRVEDEVKDQDEEAKEDEEEDLDDIEIGLLEEEVVTVLTENIVKCKNNILAHRILCQYYLLTKEYEAALPYIKNGISLIAYNIKDLGVHLPLTKREFSLDLATVYTYVDAPKDHNAALKLYDNILSGDFSNIQAKMGKGIIFIERKNWKDAMTLLTQVHEQSPNNLEVLSELSWSKAHMGYMDEALAGLDTVIKGIKGMDLRSIDFRALNLWRQAKVYIMKHASINDAKQENVKCAFKLLIQSIKILDTFAPGFSTLGDIYCHYYKDHLRAFKCYFKAFDLDAGDYTAAKYITETYASKPNWQAASSIASRLIKGEKAKAELRSNNWPFRVVGIAHLEKQEESDSIEWFQSALRVDPNDVESWVGLGQAYHACGRIEASIKVFDKAIQLRPSHTFAQYFKAISLCDVGEYLESLDILEKVCQEAATEESFQIGLVEVLMRCSLDLYSQGFLLKSVSIAKDTIERIKIIISELKCENQQVWIYLSQVLRLFIWIESKVDTLPVESLVSIFENSQFSGSEEIDSVDNIKIDTLLDSTTDDNVSIACKFLILASKYSVSDQKFTDIAGTVRASYWYNIGISELTAFITLKEPQYRDAAIFAFKKSIQLQSNTSETWIGLGIATMDINFRVSQHCFIKATALEPKATNTWFNLAMLGLKKKDTEFAQQVLNKLQSLAPQDSSPWLGMALILEEQGDIIGSSKLFAHSFILSNGRSKAAQFMYAKNVLENHINNGDDERDIETVEKLTTASIALEQFFKKSPDSQFALQCALLTLERLHHYENANELANRLIGILEKKFEKTQDERELFNFAIIKGQFARIHLGLGNFELSIENADLSQGIISESSDEKSMKTKISNHICLGLSYFFLNDFDQTLNQFQELLSISKDSKHLVVLIAKVLYDVGESDTKEIALQELTEYIATSGADLLVTLTIAAMSILDDKREDLSIILEELKALPLSKQIIDKHKDAPYLIEEITKRLYRNDTGKQVWQRSAYFFPNNLKVWERLDKNIQRRIASNGQNKVTAEEMSKLYCESKNLRSIQRGMFLCPWNVTAVKALNECF 18FFF78D7CA5A6E4 ski8_yeast Antiviral protein SKI8 YGL213C SKI8 REC103 Superkiller protein 8 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSKVFIATANAGKAHDADIFSVSACNSFTVSCSGDGYLKVWDNKLLDNENPKDKSYSHFVHKSGLHHVDVLQAIERDAFELCLVATTSFSGDLLFYRITREDETKKVIFEKLDLLDSDMKKHSFWALKWGASNDRLLSHRLVATDVKGTTYIWKFHPFADESNSLTLNWSPTLELQGTVESPMTPSQFATSVDISERGLIATGFNNGTVQISELSTLRPLYNFESQHSMINNSNSIRSVKFSPQGSLLAIAHDSNSFGCITLYETEFGERIGSLSVPTHSSQASLGEFAHSSWVMSLSFNDSGETLCSAGWDGKLRFWDVKTKERITTLNMHCDDIEIEEDILAVDEHGDSLAEPGVFDVKFLKKGWRSGMGADLNESLCCVCLDRSIRWFREAGGK C5E7B9D2268F9727 ski2_yeast Antiviral helicase SKI2 SKI2 YLR398C L8084.17 Superkiller protein 2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSEGFSSSSIQELYQSLKEITNNADVELFEDRITKLDFESTDEPKHANDIIKDRFLRPSNALPWSLLDMVQDVPHTSSPEDCSGKLDYKELLKVPDPINRTSYQFKRTGLEGKISGYKEEVDLKEVANANASNSLSITRSINHNQNSVRGSTAQLPFTPGGIPMKSVKTDSEQNGSSTMANATKLLHKDGQGLFDIPEGMNRGIKPMDSPAENEDQNGQFKELKQLNEIDNELDIRIEANEAKLKEEEKSAKSISEEIMEEATEETTADNADDAEIDELLPIGIDFGRTKPVSKSVPVKKEWAHVVDLNHKIENFDELIPNPARSWPFELDTFQKEAVYHLEQGDSVFVAAHTSAGKTVVAEYAIAMAHRNMTKTIYTSPIKALSNQKFRDFKETFDDVNIGLITGDVQINPDANCLIMTTEILRSMLYRGADLIRDVEFVIFDEVHYVNDQDRGVVWEEVIIMLPQHVKFILLSATVPNTYEFANWIGRTKQKNIYVISTPKRPVPLEINIWAKKELIPVINQNSEFLEANFRKHKEILNGESAKGAPSKTDNGRGGSTARGGRGGSNTRDGRGGRGNSTRGGANRGGSRGAGAIGSNKRKFFTQDGPSKKTWPEIVNYLRKRELLPMVVFVFSKKRCEEYADWLEGINFCNNKEKSQIHMFIEKSITRLKKEDRDLPQILKTRSLLERGIAVHHGGLLPIVKELIEILFSKGFIKVLFATETFAMGLNLPTRTVIFSSIRKHDGNGLRELTPGEFTQMAGRAGRRGLDSTGTVIVMAYNSPLSIATFKEVTMGVPTRLQSQFRLTYNMILNLLRIEALRVEEMIKYSFSENAKETLQPEHEKQIKVLQEELQTIEYKSCEICDNDIEKFLELMLAYKEATVNLMQEMVKSPSILHILKEGRLVAFRDPNDCLKLGFVFKVSLKDAVCVIMTFTKPYKLPNGEPNHLIYFPKADGYRRRNFPKFQKTDFYMEEVPVTAIEVITKRKFAAPLGKVIKKDVAALNEFNAETNNILDGKTLKEAINIEKQGLKIHQILLDRTNIRDEIFKLKSIKCPNLSQHIVPKFKAHVIKKKIEELYHLMSDQNLSLLPDYEKRLAVLKDTEFIDQNHNVLLKGRVACEINSGYELVLTELILDNFLGSFEPEEIVALLSVFVYEGKTREEEPPIVTPRLAKGKQRIEEIYKKMLCVFNTHQIPLTQDEAEFLDRKRFAMMNVVYEWARGLSFKEIMEMSPEAEGTVVRVITWLDEICREVKTASIIIGNSTLHMKMSRAQELIKRDIVFAASLYL 7CCD36CFC0DF8C32 ski7_yeast Superkiller protein 7 SKI7 YOR29-27 YOR076C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSLLEQLARKRIEKSKGLLSADQSHSTSKSASLLERLHKNRETKDNNAETKRKDLKTLLAKDKVKRSDFTPNQHSVSLSLKLSALKKSNSDLEKQGKSVTLDSKENELPTKRKSPDDKLNLEESWKAIKEMNHYCFLKNDPCINQTDDFAFTNFIIKDKKNSLSTSIPLSSQNSSFLSLKKHNNELLGIFVPCNLPKTTRKVAIENFNRPSPDDIIQSAQLNAFNEKLENLNIKSVPKAEKKEPINLQTPPTESIDIHSFIATHPLNLTCLFLGDTNAGKSTLLGHLLYDLNEISMSSMRELQKKSSNLDPSSSNSFKVILDNTKTERENGFSMFKKVIQVENDLLPPSSTLTLIDTPGSIKYFNKETLNSILTFDPEVYVLVIDCNYDSWEKSLDGPNNQIYEILKVISYLNKNSACKKHLIILLNKADLISWDKHRLEMIQSELNYVLKENFQWTDAEFQFIPCSGLLGSNLNKTENITKSKYKSEFDSINYVPEWYEGPTFFSQLYLLVEHNMNKIETTLEEPFVGTILQSSVLQPIAEINYVSLKVLINSGYIQSGQTIEIHTQYEDFHYYGIVSRMKNSKQILETNTKNNISVGLNPDILEVLVKIHNTEDFTKKQFHIRKGDIIIHSRKTNTLSPNLPNTLKLLALRLIKLSIQTHALSDPVDLGSELLLYHNLTHNAVKLVKILGTNDISINPNQSLIVEVEIIEPDFALNVIDSKYITNNIVLTSIDHKVIAVGRIACQ D93B956D02050BFA ski8-ski3-1 Interactions within the SKI complex identified by Y2H 653831 N/A 653836 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 653837 unspecified role unspecified role bait bait physical association physical association false false false Figure 1 ski8-ski2-1 Interactions within the SKI complex identified by Y2H 653831 N/A 653837 unspecified role unspecified role bait bait N/A 653838 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 1 ski3-ski2-1 Interactions within the SKI complex identified by Y2H 653831 N/A 653836 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 653838 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false 653852 653850 physical association physical association false false false Figure 1 ski3-ski2-2 Interactions within the SKI complex identified by coimmunoprecipitation 653832 N/A 653838 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 653836 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2A SKI2 with aa 444-447 mutated (DEVH -> AEVA) was equally capeable of coprecipitating SKI3 ski8-ski7-1 Interaction between Ski8 and Ski7 identified by pulldown 653832 N/A 653837 unspecified role unspecified role prey prey region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 653839 unspecified role unspecified role bait bait region 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 false physical association physical association false false false Figure 5B ski3-ski7-1 Interaction between Ski3 and Ski7 identified by pulldown 653832 N/A 653836 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false N/A 653839 unspecified role unspecified role bait bait region 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 false physical association physical association false false false Figure 5C & 6 ski3-ski8-1 Interactions within the SKI complex identified by coimmunoprecipitation 653833 N/A 653836 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c-terminal region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false N/A 653837 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 3 ski3-ski8-2 Interactions within the SKI complex identified by pulldown 653833 N/A 653836 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c-terminal region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false N/A 653837 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 653838 unspecified role unspecified role prey prey n-terminal region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false association association false false false Figure 4 Ski2 and Ski8 interaction was totally abolished in the absence of Ski3. These results suggest that the Ski2 and Ski8 interaction is bridged by Ski3, and that the N terminus (amino acids 1-279) of Ski2 is responsible for Ski3 interaction and thus required for Ski2 and Ski8 interaction. ski2-ski7-1 Interaction between Ski2 and Ski7 identified by pulldown 653834 N/A 653838 unspecified role unspecified role prey prey n-terminal region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 653839 unspecified role unspecified role bait bait region 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 false physical association physical association false false false Figure 5 Ski2 and Ski7 interaction was totally abolished in the absence of Ski3 and Ski8. ski3-ski8-3 Interactions within the SKI complex identified by glycerol gradient centrifugation 653835 N/A 653836 unspecified role unspecified role neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Stoichiometry: 1.0 N/A 653837 unspecified role unspecified role neutral component neutral component region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Stoichiometry: 1.0 N/A 653838 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Stoichiometry: 1.0 physical association physical association false false false Supp. Figure 1 MW = 380kDa (inc. tags)