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 Repression of ergosterol level during oxidative stress by fission yeast F-box protein Pof14 independently of SCF. Repression of ergosterol level during oxidative stress by fission yeast F-box protein Pof14 independently of SCF. EMBO J. (0261-4189) 2006 imex curation Tafforeau L., Le Blastier S., Bamps S., Dewez M., Vandenhaute J., Hermand D. damien.hermand@fundp.ac.be 27-Aug-2008: Contacted by JYOTI. A fission yeast cDNA library was used davet Only protein-protein interactions schpm Schizosaccharomyces pombe (fission yeast) Fission yeast fluorescence imaging fluorescence microscopy fluorescence imaging predetermined predetermined participant predetermined Tafforeau L., Le Blastier S., Bamps S., Dewez M., Vandenhaute J., Hermand D. damien.hermand@fundp.ac.be Accepted 2008-AUG-30 by DAVET. 27-Aug-2008: Contacted by JYOTI. A fission yeast cDNA library was used EMBO J. (0261-4189) 2006 davet Only protein-protein interactions imex curation Repression of ergosterol level during oxidative stress by fission yeast F-box protein Pof14 independently of SCF. Repression of ergosterol level during oxidative stress by fission yeast F-box protein Pof14 independently of SCF. EMBO J. (0261-4189) 2006 imex curation Tafforeau L., Le Blastier S., Bamps S., Dewez M., Vandenhaute J., Hermand D. damien.hermand@fundp.ac.be 27-Aug-2008: Contacted by JYOTI. A fission yeast cDNA library was used davet Only protein-protein interactions 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Tafforeau L., Le Blastier S., Bamps S., Dewez M., Vandenhaute J., Hermand D. damien.hermand@fundp.ac.be Accepted 2008-AUG-30 by DAVET. A fission yeast cDNA library was used EMBO J. (0261-4189) 2006 davet Only protein-protein interactions imex curation Repression of ergosterol level during oxidative stress by fission yeast F-box protein Pof14 independently of SCF. Repression of ergosterol level during oxidative stress by fission yeast F-box protein Pof14 independently of SCF. EMBO J. (0261-4189) 2006 imex curation Tafforeau L., Le Blastier S., Bamps S., Dewez M., Vandenhaute J., Hermand D. damien.hermand@fundp.ac.be 27-Aug-2008: Contacted by JYOTI. A fission yeast cDNA library was used davet Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip predetermined predetermined participant predetermined Tafforeau L., Le Blastier S., Bamps S., Dewez M., Vandenhaute J., Hermand D. damien.hermand@fundp.ac.be Accepted 2008-AUG-30 by DAVET. 27-Aug-2008: Contacted by JYOTI. A fission yeast cDNA library was used EMBO J. (0261-4189) 2006 davet Only protein-protein interactions imex curation Repression of ergosterol level during oxidative stress by fission yeast F-box protein Pof14 independently of SCF. Repression of ergosterol level during oxidative stress by fission yeast F-box protein Pof14 independently of SCF. EMBO J. (0261-4189) 2006 imex curation Tafforeau L., Le Blastier S., Bamps S., Dewez M., Vandenhaute J., Hermand D. damien.hermand@fundp.ac.be 27-Aug-2008: Contacted by JYOTI. A fission yeast cDNA library was used davet Only protein-protein interactions 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 Tafforeau L., Le Blastier S., Bamps S., Dewez M., Vandenhaute J., Hermand D. damien.hermand@fundp.ac.be Accepted 2008-AUG-30 by DAVET. 27-Aug-2008: Contacted by JYOTI. A fission yeast cDNA library was used EMBO J. (0261-4189) 2006 davet Only protein-protein interactions imex curation mcm3_schpo DNA replication licensing factor mcm3 mcm3 SPCC1682.02c Minichromosome maintenance protein 3 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MTELLADEVFKDRVRIFQEYLEHDTDDANVTLYQEAILRMLNMGQRRLIVNIDELRDYNRELADGVLLKPLEYVEPFDEALRNVVSTLIDPVVHKDLKDKLFYVGFRGSFGDHHVNPRTLRAMHLNKMISLEGIVTRCSFVRPKVIKSVHYCEATKRHHFKQYADATMNGGLSFQSTVYPTQDENGNPLSIEFGFSTFRDHQSISLQEMPERAPPGQLPRSIDILLDDDLVDTVKPGDRVNIVGQYRSMGSKTSGNTSATFRTVLLANNVVLLGNKPGLGNVGGGALDITDADIRNINKLARKKNVFELLSTSLAPSIYGYEYVKQAILLLLLGGTEKNLTNGTHIRGDINILMVGDPSTAKSQLLRFVLNTAPLAIATTGRGSSGVGLTAAVTTDKETGERRLEAGAMVLADRGVVCIDEFDKMSDIDRVAIHEVMEQQTVTIAKAGIHTSLNARCSVIAAANPIYGQYDIRKDPHQNIALPDSMLSRFDLLFIVTDDIDDKKDRALSEHVLRMHRYLPPGVEPGTPVRDSLNSVLNVGATNAAGVSTENVEQEVETPVWETFSSLLHANARTKKKELLNINFVRKYIQYAKSRIHPILNQATAEYITNIYCGLRNDDLQGNQRRTSPLTARTLETLIRLSTAHAKARLSSVVEVKDAKAAEKILRYALFREVVKPKRKKHKKQRLEAGEEFDSEDDNSDDMDIEESEEEMDTNMVIDSGSRRVTRSQNATSQSQESGSEIGSSIAGTAGSYNVGTSNTQLSWPSTHSTLPATSRELASSDRNINTGTSVASEVSASVSEQSTVSLPREKMSVFMARLASLTKSELFSEECASLEDVLESINNIEDDVGFSREEAIVALKEMDAQNKIMFSDNVVYRI 876051CC7DE2504A ybx1_schpo Uncharacterized protein C17D1.01 SPBC17D1.01 SPBC17D11.09 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MKAVYLDYLLRIRYTYVFNLKVSYQSERRTYGATEHFVGCLVPKAWNCLADDYTDIKTLLDLLFGVTAVIIFACTFKCFTMTEDGLTKLNLEAFLGVSGNTQELLESLGISAIPDLSNLGASNNNNSGLTFDPNDPSAAAFYIAQQVAAIEHPPPRKMTSSEKTRSENRERKKRWREQNEERNKDNDLRCRVNKKAKKLFGSEPSIEKTNWIEAEFTRRHTKRKEKERACLSQNQSSNASRANSQSLLPDLDLLATNSSANAVLQGSAINNALNALAKDPNLIHSLLTQIDFDPSKSKDGLNLGSLTDVVPPQPPQPEHELAALQEHNEAHDAAMRQYELERQQNAMLPGLASIDGLQALPHLDFSDSAVTNQSHSQSQNSLHPLISQSETHSIFSALSETPTPVSGNGNVADAPPDFSLSQVMPLDLIAPSMQPSAVSSPMSESHVQISSEMPRTGMSALPMRPRSQNVDKHRKFPYYNKRNAVTTPSSPYDGAQSGSPQFPPFELPPHNYSQAQSPNLATPSPSFSSLPDVSLPPIVKPNLMSEPTPTNSDEKSHALGFPPPPGQKIEFQRSSSKNGTAKSD 1F712A2982F59FF4 pof7_schpo F-box protein pof7 pof7 SPCC1827.08c SPCC70.11c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSTVEDQLSLELETVNKDILLKKALLHYKDAIKSEREGNLGESLNKYRLAHKVHEDVESIYRRLERLQLCKRNEEEEMLNSDASEAMLTVSSVPSPTLTENESVNESVVPNILKLPDEVLLVILENCIRDLHDLRYLSSIALTCKHFAKALRADSLYRSFCYCSCEQKEWQQSIKSIEEELVEKYQQSWKTLFLKKPRSRFDGCYISVCRYFRPGTSDTSWNQPIHLITYYRYLRLYPNSTCIVYQSSNEPNDVVRNFSVQNTSLFSPMSSSPMFSNGNVALTGSWSMTPSGEMLIVYPASQTYTYVQKLQVRGIRLKWISFYSIHNYTSFTNEMPLTHNRDYVFSRVYSYTADSESLKRG 0269BAE9AEDEB5B5 fdft_schpo Squalene synthase erg9 SPBC646.05c Farnesyl-diphosphate farnesyltransferase FPP:FPP farnesyltransferase protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSLANRIEEIRCLCQYKLWNDLPSYGEDENVPQNIRRCYQLLDMTSRSFAVVIKELPNGIREAVMIFYLVLRGLDTVEDDMTLPLDKKLPILRDFYKTIEVEGWTFNESGPNEKDRQLLVEFDVVIKEYLNLSEGYRNVISNITKEMGDGMAYYASLAEKNDGFSVETIEDFNKYCHYVAGLVGIGLSRLFAQSKLEDPDLAHSQAISNSLGLFLQKVNIIRDYREDFDDNRHFWPREIWSKYTSSFGDLCLPDNSEKALECLSDMTANALTHATDALVYLSQLKTQEIFNFCAIPQVMAIATLAAVFRNPDVFQTNVKIRKGQAVQIILHSVNLKNVCDLFLRYTRDIHYKNTPKDPNFLKISIECGKIEQVSESLFPRRFREMYEKAYVSKLSEQKKGNGTQKAILNDEQKELYRKDLQKLGISILFVFFIILVCLAVIFYVFNIRIHWSDFKELNLF BAEE7F24B6CE5D25 alp7_schpo Microtubule protein alp7 Altered polarity protein 7 Transforming acidic coiled-coil protein mia1 alp7 mia1 SPAC890.02c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSDIVSSSTDYSRRSPSSSSIGTNETDHTGFHEKRQGASSESLIPPAQRSSEESMPAPKLFPKLTSKPNPQLNLKDTLNKRVSDRLQALELNKSFDFSGTPRPMHPISHPLSQHKTPEFKHRKRNVESILTPKNPSLFSSSNAASQRGSLNTAPSNFAYSHSSSLQTSASSRPPVLSNGSFPRQTNTAPLNPPVHLKDNIRNSATPSTSQADIPTQYPINSTQKQQAKYEAEIEGYKAKLAGTYHEISVLQNTIVNVSGQLIAVNDQLQQLRSGKASTSPSTKDTNMRLVEGHNEETLALQRGKYTQEEVDKLIQERMEKVAEDLHAQYSAKHTQKINAFKANYARKYEATIQELQNQIGTAPNAPKISNSNWEEERRALKADNQTLQKQLEKAIQERQDMSDFLNNFKADMAKSDKLLMQQQSQQTGDLETLRLQLQALQEELRVEREERQQLIQMSEDLVIAMDQLNLEQKS 22B4216C196E310C 22B4216C196E310C pof14_schpo F-box protein pof14 pof14 SPAC13D6.01 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MLQFSFQNCKTHTCYMNTSVNEHLDQDESFLRILIDTRKKIRSSYFKPQTFDEFVHCLARVRAMKRLVSICSNFDEEDNWNGVWNTLVVDGDSHASAMIDYEGLLDKCSLSRNLLELINDYAEYTTQVLPFRKIPSSDNLDSSLNLTVASPKSNLYYRYSSESPKYAKILDCPDEILQLIFSYCYDASYIEKLPFAFSYRKQRHTLIHDLPNTCLRFKKILSPRNVSFWKRLLKVHKKNPNSAVTCSESINTSAVAKFTDIPTIIPIFLDPSYQSYAAKTIHSDTSSLASSIIQTGDGTQSCDESTYIELACNLVGRCVLCNRIPKLTKFKDCITSFYRPSVATNICDQCLEAIIDFYEPVSRYKFDVMKDRLGVGYISRPGQKFPSWLSEHVQLARDEEKAFKSIYHSQGEFARSLFRLFRAQLAELCEQ 132FB0919646AC9F pof6_schpo F-box protein pof6 pof6 SPCC18.04 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MKPSEAREHAIRKIKEYLERRHKQQFKALFGCLTINIYLKIFTLISTPDLCNCRLVCRKFQQLCDYNSIYVKKLLLMNAWDVTLSRKLYYESQDGMSQANMSSNTSKGFHLQSSDKKYADSDRPKLSSSSLLSLPDQKIPFPVDYSTVQGKQVVLTVLDCVSQRVEFARLDYIKVWRALAPIYRNLAYASNTIDPIAFSAFRTPEEQSKVLKYLIRFGNSYPYGTYRQAMQNAILDMASLFEQACLDEFELGLHSRNLDLLRKFSHVLHDFSGPNAYVSMYLAKQTDFVRSFFHFDPYSLFISNNLEEIHINWNILESVVNDTIKLLESESKFSEATLPEPELVQVPYAKDILGNSLKDYVISICEHIGEEETELFLVFISGFYGLCKKFFSIPNGPALVDTIFQPQIDIFISQELHYFKTVGWSLVDQWDQKLEEKEDATECFFYKNVSQNTAKNNFLETFKNVMLLPVSLFTIPSENNSASNLAEKAIEQKEEEDPELSKLDAARFVPANIYVSKDRLKHLPTTELAAQAAVLDSKLEGISTMFSLELALKIVHLCKVSLARAKVFMGTSVPQDDDIKGLSKDLFVQLLRELGQGHLKHGFDRAIEHLSSFDPRRDFSSNTVEPVVKFLELINVGDMIQQMMDSFFNEEMSPICVKDDMFDPAIAEKKKFEQLLDERAAFGLHKGINVLIEHADFLLETKTPMNLFSDQTIGSITNTIEPTAAAKNVVQFLGFHMRILVGRADHEILDVFYKEVGMRLFDSLTRYIKSHKFSVDGGLKLLSDCNLYYEFIHSLHQSSLLPYFKTLKEIAHLFIIDGKNAEEIGKLATDTSRFSSAFHTEEVYEILHSRIDWLNIKYEVDKVIHGLACCIM 0F92F9824F579D6C pmgy_schpo Phosphoglycerate mutase gpm1 SPAC26F1.06 Phosphoglyceromutase MPGM BPG-dependent PGAM protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MTTEAAPNLLVLTRHGESEWNKLNLFTGWKDPALSETGIKEAKLGGERLKSRGYKFDIAFTSALQRAQKTCQIILEEVGEPNLETIKSEKLNERYYGDLQGLNKDDARKKWGAEQVQIWRRSYDIAPPNGESLKDTAERVLPYYKSTIVPHILKGEKVLIAAHGNSLRALIMDLEGLTGDQIVKRELATGVPIVYHLDKDGKYVSKELIDN 4B937820E2FA5C15 lcf2_schpo Long-chain-fatty-acid--CoA ligase 2 lcf2 Fatty acid activator 2 Long-chain acyl-CoA synthetase 2 SPBP4H10.11c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MDIKDYYCVETKDSKQPGESYIYRSIHSAGKLLDTPEDGVSTVYDLLCTAAKNHGEKQAMGSRKLIKENREEREIIRKVDNEEVVEKKLWTTYELGPYEYISFNKVYEIALALGSGLVASGITSETTMLFFAATSAKWFTTAQGCSSQAIPIVTAYETLGEDGIYTSLDECKSRAIFTDPNLIPKLLGPLKQSTWVKLIVCSSTPSEDLVELVKSTAPDVEIITYDNLLSLGKEKPQPPHPPKADDICCYMYTSGSTGKPKGVVLLHRNIIAALGGINRILSQHINVKDYVLAYLPLAHIFEFIFEMCCLYWGGVLGYASPRTLTDASVVNCKGDLTEFRPTVLIGVPAVYELIKKGILAKVSSMPAHRQKVFSGSLSLKQYLIERNLPGSAALDAIVFNKVKAATGGRLRYCISGGAALAASTQAFLSSCICPVLPGYGLTETCAGSFVLSPEQWHLYANTVGFPIPSIEFKLVDIPDLGYYTDSSPPRGEVWIRGPAVCNGYLNRPEDNKAAFTEDGWFKTGDVGEIAKGNTLRLIDRKKNIVKSLNGEYIALEKIEAQFFTSPLVSNVCAYADVNHAKPVVIVNPDENGLRTYLTKNSGSSFNGNPNDTLTNLCKDSGVQHLILKELINIGKQQRLASIEIPEGVVLSDFEWTAENNFLTASRKVKRQVIVAHYSDEIQKAYSKKH 5F80C06F6EA92F87 pop2_schpo WD repeat-containing protein pop2 Proteolysis factor sud1 pop2 sud1 SPAC4D7.03 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSLSRCPTDNSSSRINSSVPLINSSSPATPPESFDPQVFPSSLIHGDNLLPQDDQIASDPRSESNSCNGNTSSSLPCTDSYQYPLKHSCTPSFLRKFNESIENVSYKCLDHSPPDSVPGDFSISLVPQRNFLYSHSSLPPKIISIDRNNRIKLDNSISSNSDNFPPSPKVDTSNTVSPGSKPISEDLEDLNLQSIVQTFEDLPEGIQSYAFFQLLRSCNRQSMRLLLNECEPLLKKDILSNLPFSIVQSILLNLDIHSFLSCRLVSPTWNRILDVHTSYWKHMFSLFGFQINENDWKYANPNLNRPPFLHNDQISDDYFPEIFKRHFLNRKRWLFPSIPPSHLSFPIHVPNFMITSLLLHKDRIITTSGSGTIQIHNAITGVLEARLEGHKEGVWAVKIHENTLVSGSIDKTVRVWNIEKAKCTHIFRGHISIIRCLEILVPSRLIRHGVEIVEPDQPYIVSGSRDHTLRVWKLPKNTDPPYLPDNTNSIDRWEKNPYFVHTLIGHTDSVRTISGYGDILVSGSYDSSIRIWRVSTGECLYHLRGHSLRIYSVLYEPERNICISGSMDKSIRVWDLSTGTCKYVLEGHDAFVTLLNVFQNRLISGSADSTIRIWDLNTGKPLMVLPSNSGYISSFVSDEHKIISGNDGSVKLWDVRTGKLLRFLLTDLTKIWHVDFDAMRCVAAVQRDDQAYLEVINFSGSRP 0336A0568C152396 pgk_schpo Phosphoglycerate kinase pgk1 SPBC14F5.04c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSLSTKLAITDVDLKGKNVLIRVDFNVPLDGDRITNNARIVGALPTIKYALEQQPKAVILMSHLGRPNGARVAKYSLKPVAAELSKLLGKPVKFLDDCVGPEVEKACKEAKGGEVILLENLRFHIEEEGSAKVDGKKVKADASAVEAFRKSLTSLGDIFVNDAFGTAHRAHSSMVGVDLPRVSGFLMKKELDYFSKALENPARPFLAILGGAKVADKIQLIDNLLDKVNRLIICGGMAFTFLKVLNGMKIGDSLFDEAGSKNVESMMAKAKKNNVEVFLPVDFVTADKFDKDAKVGSATAEEGIPDGWMGLDCGPKSSAKFAEVITTSKTIVWNGPAGVFEFDNFAKGTKSMLDACVKTCEAGNVVIVGGGDTATVAKKYGKEDALSHVSTGGGASLELLEGKALPGVVALSSK F40D706D5487EB4E cul1_schpo Cullin-1 cul1 pcu1 SPAC17G6.12 Cell division control 53 homolog protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MTTLNTNDKDLPIVKKYDSLNGTWDFLKTGVSQIFERLDEGMTITKYMELYTAIHNYCADASKTITVDNFNDQTANVLGEALYNNLVLYLEEYLARLRKECISQTNHEEQLAAYAKYWTRFTTSARFINHLFGYLNRYWVKLKNRFTETLVYDIYTLCLVSWHHHVFSHIRDSLLQNLLYMFTKKRLYEPTDMKYVEVCVDSITSLSFDKTDMTKPNLSSYKTFFETNFIENTKNFYAKESSEYLASHSITDYLKKAEIRLAEEEELVRLYLHESTLKPLLEATEDVLIAQHEEVLHNDFARMLDQNCSEDIIRMYRLMSRTPNGLQPLRQTFEEFVKRSGFAAVAKIVPQVGGEADVDPKEYMEMLLSTYKASKELVNTAFHGDTDFTKSLDTAFRELVNRNVVCQRSSSRSPELLAKYADSILRKSNKNVDIDDVEDCLSSIIIIFRYVEDKDVFQNFYTKLLAKRLVNGTSNSQDAESSMLSKLKEVCGFEYTSKLQRMFQDISLSQEITEAFWQLPQSRAGNIDFSALVLGTSFWPLSPNNVNFHLPEELVPLYEGFQNYYYSCHNGRKLSWLFHLSKGEIKARINPQTNVTYVFQVSTYQMGVLLLYNHRDSYTYEELAKITGLSTDFLTGILNIFLKAKVLLLGDNDKLGDPNSTYKINENFRMKKIRVQLNLPIRSEQKQESLETHKTIEEDRKLLLQSAIVRIMKARRTLKHVVLVKETIDQIKSRFTPKVSDIKQCIDMLIEKEYLERQGRDEYIYLA 458CA31B5D97FAA9 ecm33_schpo Cell wall protein ecm33 ecm33 SPAC1705.03c SPAC1F2.01 SPAC23H4.19 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MLFKSFALTLLFAAARVQAASNCSSGPYNISAQGTLDELNSCTVLNGDLYISDAGNSGITTLTVNGIESVQGDVVVSDGQYLTSLSFPSLKNVSGAFNVNNMIRMNNLATPELTSVGSLNLAVLPNLQELQFNAGLSDSDSVVIDDTQLQAIDGISLDSVTTFQVTNNRYIQEITMEGLESAQNIQISANSKGVSVNFSKLSNVTTATFDGISNVFIGNLKSAAGNLYFSNTTLDNISVPYLTEIGQSFAVLYSPELTSLNFPNLTTVGGGFVINDTGLTSIDGFPVISEIGGGLVLLGNFSSIDMPDLSDVKGALTVETKATNFTCPWSNDDSVIKGDDFTCQGSVATISATSSYDLSSTVSATSGSATSATGSATTTSYSSDSSASSSSSSSHESSAASNGFTAGALVLGSLLVAALAM A349C6D8501D8A29 ral2_schpo Protein ral2 ral2 SPBC21.05c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSEVKRNISLRNSTNIFTSTFSLSSNNVPKPLIGESVIKYGDEAFVYGGRDALNAQLVNDMYVVDLNTCSWKQVEYQGNQKPIPRYFHSGDLWNNKLIFFGGMGFNDDTKCLYVLNDIDIYDIETKQWSHIPGMITENQTNDDAKEVNGSDVDEKSKHLYPSARYGHLHCVLDHYLIIFCGQDLSNSYIEEINIFDLDSGKWVFKSLFNHHCGIYRSNCVVINKDSEFLQMCRPINTTQDSNEHSIGSLFFYLNYNFVNVKRQVIYLELFELDTAESEKKSAALAKDNNQSFRFLELDVTEKFLSSAMPPGLRFPAVNILGDNLILSGIYLTSSRQAFVLWVYSLDKELWLQLDMLGVLNHGSWFKCLVLDCTNRFVVFGNKTRKLTQDYNLRQSNYDHIVFIELEGYGVYRKPQMGRVTERSEQLGKLLLNGISDMEILTIERMHIPCLSRMLYKRWPAFQKIMDRAVEKNQEAFQAEVSQLGPQLTDLPFSSIHSTGSRALYMPYSFETCSAFLHYIYCGTLNGSYCTAKNLCNLLILCKGFEGLETFFAYIVHLLHGVLNRNNVKLIYETAALTGAKGLQLRALRRIARIEQGGTAISPTSPLPNLDD 1C424B2D31F00AAB eif3a_schpo Eukaryotic translation initiation factor 3 subunit A tif32 SPBC17D11.05 eif3a Translation initiation factor eIF3, p110 subunit Eukaryotic translation initiation factor 3 110 kDa subunit protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MAPPQGKPENVLRLADELIALDQHSSALQSLHETIVLKRSRNAQGFSLEPIMMRFIELCVHLRKGKIAKEGLYTYKNAVQNTSVTAIENVVKHFIELANKRVQEAQEKADKISVEYVDDLEATETPESIMMSLVSGDLSKSRTDRALVTPWLKFLWDAYRTVLDILRNNARLEVMYQLIANSAFQFCLKYQRKTEFRRLCELLRSHLGNASKFSNAPHSINLNDAETMQRHLDMRFSQLNVAVELELWQEAFRSIEDIHSLLTFSKRAPAAVMLGNYYRKLIKIFLVCDNYLLHAAAWNRYFTFTNVQKPATANFVILSALSIPIIDANKLSGPSIEAEDAKSKNARLALLLNLSKTPTRETLIKDAISRGVLSFCDQAIRDLYQILEVEFHPLSICKKLQPIIKRLAESNDTAQYIRPLQQVILTRLFQQLSQVYDSISLKYVMDLATFEEPYDFNPGQIEKFIMNGNKKGAFSIRLNHIENSISFSSDLFSNPIKSSDSVSLQSTPSELITSQLTRIAKSLSSVLMRFDTDFCLLRKQQAEAAYERAQAGVEQERKAVIAQRSLLELRRGQADTLATQREAELAAQRALKQKQESEAESLRVQEEINKRNAERIRREKEAIRINEAKKLAEELKAKGGLEVNAEDLEHLDADKLRAMQIEQVEKQNKSMNERLRVIGKRIDHLERAYRREAIPLWEEDAKQQAEHDREIFYEREKQRKEVQERKHEQAIKDKKAFAQFASYIHAYKQNIDDERDKAYQEAYAKAKNVIDAERERQRKEIFEQKLAEAIREAEEEAARAAEEEANRELHEQEEAQKRAIEERTRAAREAKEREQREMAEKLERQRRIQQERDEEISRKLAEKAAARRANIGASSPSPGAWRRGGASAGGVSRDSPRYSRGGYSRGSVPPRETLAPSKGAYVPPSRRNQQQQ 120EEDBC5C43DBC9 ei2ba_schpo Translation initiation factor eIF-2B subunit alpha tif221 SPCC11E10.07c eIF-2B GDP-GTP exchange factor subunit alpha protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MVESDSSGIVRHSQGEFDIVQVYKKFLQDDPEITMPVAAIEALVQLLSRSQAKTISEFMDILQNGSNTLKEGVQNNISLSAGCDIFQRFVTRSLHDVGDFEQCKRHLVENGKLFIQRARACRQRIAHLGYPLIRDGSVILTHGFSRGVAAVLLAAAKRHVRFKVFVTESRPSGSGCLMTRTLKNACIPTCMVLDSAVSFTMNRVDLVLVGAEGVVENGGLINQIGTFQLAVFAKHAHKPFYAVAESHKFVRMFPLSQYDIPFSRPILEFDDPSPETVHPEPEPIPTPSDAIHNELIMNEEQIRNNPTLDVTPPEFVSGLITDLGIIDSKSGVSEELIKLYL 6FAACEBEBD50995A 6FAACEBEBD50995A ydri_schpo Uncharacterized protein C16E8.18 SPAC16E8.18 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MEVTSFILNATFKEFACFGNNYLIILPGIMLERNVFRHLNYSTNSICSHYQFFGGHYESFELLVVIVYYFSHVGSFSLAEIYRITWDKRIVLYGTTTTLVYCSEGSD C47FC7F6DF29B4A6 ap1_schpo AP-1-like transcription factor pap1 caf3 SPAC1783.07c Caffeine resistance protein 3 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSGQTETLSSTSNIPIAKAEPEQSADFSASHKKRGPVSDRSSRRTSSEEVDLMPNVDDEVDGDVKPKKIGRKNSDQEPSSKRKAQNRAAQRAFRKRKEDHLKALETQVVTLKELHSSTTLENDQLRQKVRQLEEELRILKDGSFTFEMSLPHRNPSLSSLPTTGFSSNFAHMKDGISPQSNLHLSPNSIEKPNMHQNVLHNDRSADNLNHRYQVPPTLVDSNSAQGTLSPETPSSSDSPSNLYLNYPKRKSITHLHHDCSALSNGENGEDVADGKQFCQKLSTACGSIACSMLTKTTPHRASVDILSNLHESTVSPPMADESVQRSSEVSKSIPNVELSLNVNQQFVSPFGGTDSFPLPTDTGLDSLFEPDSAIENSHLKNVVMEPELFQAWREPAESLDKEFFNDEGEIDDVFHNYFHNSNENGDLITNSLHGLDFLENANESFPEQMYPFIKHNKDYISNHPDEVPPDGLPQKGKHDTSSQMPSENEIVPAKERAYLSCPKVWSKIINHPRFESFDIDDLCSKLKNKAKCSSSGVLLDERDVEAALNQFN D9E7FEA0A286B4BE skp1_schpo Suppressor of kinetochore protein 1 skp1 psh1 sph1 SPBC409.05 P19/Skp1 homolog protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSKIKLISSDNEEFVVDQLIAERSMLIKNMLEDVGEINVPIPLPNVSSNVLRKVLEWCEHHKNDLYSGTEEESDIRLKKSTDIDEWDRKFMAVDQEMLFEIVLASNYLDIKPLLDTGCKTVANMIRGKSPEDIRKTFNIPNDFTPEEEEQIRKENEWAEDR 07EA1E97A3556E78 g6pd_schpo Glucose-6-phosphate 1-dehydrogenase zwf1 SPAC3A12.18 SPAC9.01 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSSANLSIKENGAMVVFGASGDLSKKKTFPALFSLFSEGRLPKDIRIVGYARSKIEHEDFLDRITQNIKIDEEDSQAKEKLEEFKKRCSYYRGSYDKPEDFEGLNSHLCEREGDRSTHNRIFYLALPPDVFVSVATNLKKKCVPEKGIARLVIEKPFGVDLKSAQELQSQLAPLFDEKEIYRIDHYLGKEMVQNLVHLRFCNPVISHLWDKNSISSVQITFKEPIGTEGRGGYFDSSTIVRDIVQNHLVQILTLLTMETPTTFSADDLRDEKVKVLRRTRLGDLKDIVLGQYVKSKDGKKPGYLDDETVPKGSRCPTYSAIPCFIDTERWRGVPFLLKAGKAMDIGKVEIRVQFKAAANGLFKDAYHNELVIRVQPDEAIYFKMNIKQPGLSEAPLLTDLDLTYSRRFKNMKLHEAYEALFLDAFAGDQSRFARIDELECAWSLVDPLLKYMEEEKPVPEPYEYGSDGPECLYSFLKKFGYIYDSPDYYDYPVMSVPSDH 6706DD0847762A22 mug64_schpo Meiotically up-regulated gene 64 protein mug64 SPAC56F8.05c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast METFKTHITGQFNATVSSITPFAKKTTHYLRTKIGSRVEELSLPEDYVELEQQVDSLKEAYNLVLPIVETVEVDGYDYPTNFRDSITDFGKTVSGKVRNLGNLTPLEQTPLASVGKNLEEKEAAAKPSRTLYNAISRAASEATTKMGAGNPLSSAFGQISVLEEKVGNLQGERDSAISKNFCEQVRAVLYPRFAEAHRVKADVQDKRLQLEMAKLDVESAKPENLEHCKSAVRSAEDELNGAIEHAKILYEQILNKDYNTDLLRSIIKNQLKFHQDAAAALSDITI 2EC915AC7CED234B tfb3_schpo RNA polymerase II transcription factor B subunit 3 RNA polymerase II transcription factor B 38 kDa subunit RNA polymerase II transcription factor B p38 subunit CDK-activating kinase assembly factor MAT1 homolog RING finger protein pmh1 pmh1 mcr1 tfb3 SPBC776.18c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MDDEGARKVDEKCPLCQADRYLNPNMKLLINPECYHKMCESCVDRIFTTGPAQCPTPGCNKILRKAKFREQTFEDAQIEREVDVRKRISRIFNKGQQEFDSLQAYNDYLEEVEILTFNLIYKIDVEETEEKVKQYEKQNRDSIAANSARAAAEARILAQNEILLKRQKQEAREAAIREHQKEKERREQVEQQIIFDLATSGKDPNKIIQLSDSLKKQQENIASSVSNISRSSSILLSDVQQVAEDTTPFSPLAGEKDGSKYFSYSKNTYQDLYLEKVSHEPGRKCGFRIEDCHLRCLYEAFSGIDYDLESLKKLEVAS FF8472C61268565B pof10_schpo F-box/WD repeat-containing protein pof10 pof10 sbp2 SPBC1703.06 Skp1-binding protein 2 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MKSEPTSLDFTSSNLRRMNRDHSSNNTNRTVLNLPKEILIIIFSFLDPRSLLSAQCTCKYWKKLLSDDLSWRTAFFHHFAGDQSQIFSPLGNGTWRQEYLLRSTITRAYEKGKGQTVQYDCRVGQLTNLYYDFSSGRLYSGNWLTGTISVSDPTTGKVERSLLHASTDGSFTHGLSTMTLGKQIFGFGFMDGRVGVILMSRQAETPRKFRYCLDSHADSVTCIDALTGDLPPTGEIGMVTGSDDGSVHCWDVKTGVSLQSFQFRSSQILSLCFRPKYKMLLVDTFNYELNSYQLYLIPGYARSRKNEQPILLSSRKCVLTDEEEPPCLMTADCCAGVAFLSRGAPKNCICRVSFKEFLEKNDNVGVQTSSIPLNGKPTSISLDTNDRVLSKSTPGRGARLLAVGDENGLVYVVNTRTEDPNKAILRTITAYSNFPITDIYLNEVAMVVGSASGYCGVYDTVTGNFLKKIASARNAARREPINCILLDSNPLSLKGVITMSKHVKSWSYTIPKPFVNKRSKVLPLRPSVTHDNLSKSSDYSKNEVEREIMLGLDQIAQERREKMEARQKFEQHFGEGLVGLSEEEIIAYVTMLSQEEEAKRMVQLSMDVDKIEEDFKENDEQATSSLNALSSNHEPPQEQANVAELNEQEQIELAMRLSLMEM 44061C20A61F602C rm15_schpo 54S ribosomal protein L15, mitochondrial mrp15 mrpl15 SPBC1539.01c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MLTRHCNRLGLQIENKFVFRSSSWNCVRRIGKIACNENKYRYEMTSTEEDIDSFFSRVFGEKNYISDKSLREQIVTHKSYKHGLKRYNEPFVSVGNAVIDVCLLNHYLESGNFSTIPSYKDLVNLRSPARLAEVAKKWNPETATQCILPLKGPNSPSSSSFEKIYSSIVSGLVAVVYFQKGGVEAMNFCKKSIISDLTKDFVRPNTVHNQ FCF2BB08A626D927 yfy2_schpo Uncharacterized N-acetyltransferase C9.02c SPAC9.02c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MVNIRNIERSEVEEVSKLEAICFPEAERASFARIKDRVKRAPEIQLGLFAPTQHTSTLIGHVLTSRTGSETVTHESMSKHDSKGENIAIHSLSIHPAHRKNGYAKKLIEELQFRCQHTITPKPSRMILISHKPLVPFYESLGFKNLGPSECKFGDETWYDMTHKI 3E0BA839B887855D skp1-pof10-4 2 23 unspecified role unspecified role bait bait 27 unspecified role unspecified role prey prey physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Pof10 was found to interact with Skp1. skp1-pof14 2 10 unspecified role unspecified role prey prey 23 unspecified role unspecified role bait bait physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Pof14 was found to interact with Skp1. skp1-zwf1 2 24 unspecified role unspecified role prey prey 23 unspecified role unspecified role bait bait physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Zwf1 was found to interact with Skp1. skp1-pap1 2 22 unspecified role unspecified role prey prey 23 unspecified role unspecified role bait bait physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Pap1 was found to interact with Skp1. skp1-yrdi 2 21 unspecified role unspecified role prey prey 23 unspecified role unspecified role bait bait physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Yrdi (SPAC16E8.18) was found to interact with Skp1. skp1-pof6-3 2 23 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Pof6 was found to interact with Skp1. skp1-pof7-1 2 23 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Pof7 was found to interact with Skp1. skp1-mcm3 2 23 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Mcm3 was found to interact with Skp1. skp1-pop2-1 2 23 unspecified role unspecified role bait bait 14 unspecified role unspecified role prey prey physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Pop2 was found to interact with Skp1. skp1-mrpl15 2 23 unspecified role unspecified role bait bait 28 unspecified role unspecified role prey prey physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Mrpl15 (SPBC1539.01c) was found to interact with Skp1. skp1-ral2 2 18 unspecified role unspecified role prey prey 23 unspecified role unspecified role bait bait physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Ral2 was found to interact with Skp1. skp1-alp7 2 9 unspecified role unspecified role prey prey 23 unspecified role unspecified role bait bait physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Alp7 was found to interact with Skp1. skp1-ybx1 2 23 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Ybx1 (SPBC17D11.09) was found to interact with Skp1. skp1-pgk1 2 23 unspecified role unspecified role bait bait 15 unspecified role unspecified role prey prey physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Pgk1 was found to interact with Skp1. skp1-lcf2 2 13 unspecified role unspecified role prey prey 23 unspecified role unspecified role bait bait physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Lcf2 (SPBP4H10.11c) was found to interact with Skp1. skp1-yin3 2 23 unspecified role unspecified role bait bait 17 unspecified role unspecified role prey prey physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Yin3 (SPAC1705.03c) was found to interact with Skp1. skp1-ei2ba 2 20 unspecified role unspecified role prey prey 23 unspecified role unspecified role bait bait physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Ei2ba/tif221 was found to interact with Skp1. skp1-yfy2 2 23 unspecified role unspecified role bait bait 29 unspecified role unspecified role prey prey physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Yfy2 (SPAC9.02c) was found to interact with Skp1. skp1-pmh1-2 2 23 unspecified role unspecified role bait bait 26 unspecified role unspecified role prey prey physical association physical association Supplementary Table 1 We screened 107 clones from a fission yeast two-hybrid cDNA library using Skp1 as bait. Ninety-four positive candidates representing 19 proteins (Supplementary Table I) were isolated. Pmh1 was found to interact with Skp1. pof14-skp1 2 23 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association Supplementary Table II, Figure 2A To gain insight into the function of Pof14, a second twohybrid screen with Pof14 as bait was performed and 61 candidates representing five proteins were isolated (Supplementary Table II). Two clones were recovered at similar high frequency: Skp1, as expected, and Erg9 (Figure 2A and B). Erg9 is the squalene synthase, a membrane-bound enzyme that plays a pivotal role in sterol metabolism. Analysis of overlapping interacting clones revealed that the C-terminal part of Erg9 was sufficient for interaction with Pof14 (Figure 2B). Skp1 was found to interact with Pof14. pof14-erg9 2 8 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association Supplementary Table II, Figure 2A, 2B. To gain insight into the function of Pof14, a second twohybrid screen with Pof14 as bait was performed and 61 candidates representing five proteins were isolated (Supplementary Table II). Two clones were recovered at similar high frequency: Skp1, as expected, and Erg9 (Figure 2A and B). Erg9 is the squalene synthase, a membrane-bound enzyme that plays a pivotal role in sterol metabolism. Analysis of overlapping interacting clones revealed that the C-terminal part of Erg9 was sufficient for interaction with Pof14 (Figure 2B). Erg9 was found to interact with Pof14. pof14-gpm1 2 12 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association Supplementary Table II To gain insight into the function of Pof14, a second twohybrid screen with Pof14 as bait was performed and 61 candidates representing five proteins were isolated (Supplementary Table II). Gpm1 was found to interact with Pof14. pof14-mug64 2 10 unspecified role unspecified role bait bait 25 unspecified role unspecified role prey prey physical association physical association Supplementary Table II To gain insight into the function of Pof14, a second twohybrid screen with Pof14 as bait was performed and 61 candidates representing five proteins were isolated (Supplementary Table II). Mug64 was found to interact with Pof14. pof14-tif32 2 10 unspecified role unspecified role bait bait 19 unspecified role unspecified role prey prey physical association physical association Supplementary Table II To gain insight into the function of Pof14, a second twohybrid screen with Pof14 as bait was performed and 61 candidates representing five proteins were isolated (Supplementary Table II). Tif32 was found to interact with Pof14. skp1-pof14-1 4 10 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 23 unspecified role unspecified role bait bait physical association physical association Figure 1A, 1B However, dissection of the protein sequence with the help of the Blocks software (Henikoff et al, 2000) revealed the presence of a putative F-box between residues 172 and 214. To test this possibility, a Gal4-AD (Gal4 activation domain) fusion protein bearing deletion of the first 14 residues (172�186) of the predicted F-box was constructed and tested against Skp1 in the two-hybrid system. Figure 1A� C shows that in spite of similar expression levels, only the full-length protein retained ability to interact with Skp1. Only the full-length Pof14 protein retained ability to interact with Skp1, indicating that an intact putative F-box is required for this binding. pof14-skp1-2 3 10 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 23 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association Figure 1D. We checked the ability of Pof14 to form an SCF complex in vivo. Co-immunoprecipitations from fission yeast extract revealed that Pof14 forms a complex with the SCF core components Skp1 and Pcu1 (Figure 1D). Pof14 protein interacts with Skp1. pof14-cul1 3 16 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 10 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association Figure 1D. We checked the ability of Pof14 to form an SCF complex in vivo. Co-immunoprecipitations from fission yeast extract revealed that Pof14 forms a complex with the SCF core components Skp1 and Pcu1 (Figure 1D). Pof14 protein interacts with Cul1/Pcu1. pof14-erg9-1 4 10 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Figure 2A, 2B. To gain insight into the function of Pof14, a second twohybrid screen with Pof14 as bait was performed and 61 candidates representing five proteins were isolated (Supplementary Table II). Two clones were recovered at similar high frequency: Skp1, as expected, and Erg9 (Figure 2A and B). Erg9 is the squalene synthase, a membrane-bound enzyme that plays a pivotal role in sterol metabolism. Analysis of overlapping interacting clones revealed that the C-terminal part of Erg9 was sufficient for interaction with Pof14 (Figure 2B). The C-terminal part of Erg9 was sufficient for interaction with Pof14. pof14-erg9-2 3 10 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 8 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association Figure 2C Pof14 and Erg9 could be co-immunoprecipitated from fission yeast extracts (Figure 2C), supporting the fact that the two-hybrid interaction is physiologically relevant. However, neither Skp1 (Figure 2C) nor Pcu1 (data not shown) could be co-immunoprecipitated with Erg9, raising the possibility that two Pof14-containing complexes might coexist in vivo, one of them being a canonical SCF and the other one containing Erg9. Erg9 was able to interact with Pof14 but not with Skp1. skp1-erg9 3 23 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 8 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association Figure 2C Pof14 and Erg9 could be co-immunoprecipitated from fission yeast extracts (Figure 2C), supporting the fact that the two-hybrid interaction is physiologically relevant. However, neither Skp1 (Figure 2C) nor Pcu1 (data not shown) could be co-immunoprecipitated with Erg9, raising the possibility that two Pof14-containing complexes might coexist in vivo, one of them being a canonical SCF and the other one containing Erg9. Erg9 did not interact with Skp1. pof14-erg9-3 1 10 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 8 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association Figure 2D As reported for the budding yeast homologue of Erg9 (Kumar et al, 2002), SpErg9 associated with vesicle-like and ER (including a nuclear rim) structures (Figure 2D, left panel), consistent with the presence of a conserved transmembrane domain in Erg9 (Figure 2B; Robinson et al, 1993). Pof14 displayed a very similar localisation (Figure 2D, central panel), and colocalisation was confirmed (Figure 2D, right panel). Erg9 and Pof14 colocalized at vesicle-like and ER structures. pof14-skp1-3 3 23 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 10 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association Figure 3D. However, it is not likely to be the case because when immunoprecipitating Skp1 in the presence or absence of stress (40 min, 0.2mM H2O2), a similar amount of Pof14 was co-precipitated (Figure 3D), although the total Pof14 concentration was elevated as shown above (Figure 3B). This rather suggests that the SCF might be saturated when Pof14 is induced, therefore leading to a stabilisation of the Pof14 pool present in the cell. Pof14 could then only reach its normal level after transcriptional induction has been turned off. Pof14 protein interacts with Skp1 even under stress conditions (being exposed to 0.2mM H2O2/40mins). pof14-erg9-4 3 8 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 10 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association Supplementary Figure 1 The induction of pof14 transcription by hydrogen peroxide prompted us to analyse the Pof14�Erg9 interaction under oxidative stress conditions (H2O2 0.2mM for 40 min). An increased amount of Pof14, proportional to its elevated level in total cell extract, was co-precipitated by Erg9, suggesting that after induction, the newly synthesised Pof14 proteins bind to Erg9 (Supplementary Figure 1). Erg9 was able to interact with Pof14 under stress conditions (0.2mM H2O2, 40mins).