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/ Nucleotide excision repair in yeast is mediated by sequential assembly of repair factors and not by a pre-assembled repairosome. Nucleotide excision repair in yeast is mediated by sequential assembly of repair factors and not by a pre-assembled repairosome. J. Biol. Chem. (0021-9258) 1996 imex curation Guzder SN., Sung P., Prakash L., Prakash S. davet Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast pull down pull down western blot western blot Immuno blot Immuno blot Guzder SN., Sung P., Prakash L., Prakash S. Accepted 2009-SEP-24 by DAVET. J. Biol. Chem. (0021-9258) 1996 davet Only protein-protein interactions imex curation Nucleotide excision repair in yeast is mediated by sequential assembly of repair factors and not by a pre-assembled repairosome. Nucleotide excision repair in yeast is mediated by sequential assembly of repair factors and not by a pre-assembled repairosome. J. Biol. Chem. (0021-9258) 1996 imex curation Guzder SN., Sung P., Prakash L., Prakash S. davet Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Guzder SN., Sung P., Prakash L., Prakash S. Proteins first bound, non-specifically, to Ni-Agarose, then eluted from a Sephacryl S-300 HR column. Accepted 2009-SEP-24 by DAVET. J. Biol. Chem. (0021-9258) 1996 davet Only protein-protein interactions imex curation Nucleotide excision repair in yeast is mediated by sequential assembly of repair factors and not by a pre-assembled repairosome. Nucleotide excision repair in yeast is mediated by sequential assembly of repair factors and not by a pre-assembled repairosome. J. Biol. Chem. (0021-9258) 1996 imex curation Guzder SN., Sung P., Prakash L., Prakash S. davet Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Guzder SN., Sung P., Prakash L., Prakash S. Proteins first bound, non-specifically, to Ni-Agarose, then eluted from a Sephacryl S-300 HR column. Eluted complex then immunoprecipitated. Accepted 2009-SEP-24 by DAVET. J. Biol. Chem. (0021-9258) 1996 davet Only protein-protein interactions imex curation Nucleotide excision repair in yeast is mediated by sequential assembly of repair factors and not by a pre-assembled repairosome. Nucleotide excision repair in yeast is mediated by sequential assembly of repair factors and not by a pre-assembled repairosome. J. Biol. Chem. (0021-9258) 1996 imex curation Guzder SN., Sung P., Prakash L., Prakash S. davet Only protein-protein interactions in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Guzder SN., Sung P., Prakash L., Prakash S. Accepted 2009-SEP-24 by DAVET. J. Biol. Chem. (0021-9258) 1996 davet Only protein-protein interactions imex curation ssl1_yeast General transcription and DNA repair factor IIH subunit SSL1 SSL1 RNA polymerase II transcription factor B subunit SSL1 Suppressor of stem-loop protein 1 YLR005W protein protein yeast Saccharomyces cerevisiae Baker's yeast MAPVVISESEEDEDRVAITRRTKRQVHFDGEGDDRVDQQQQQHSSSHRDRDKHVQRKKKKRLSNRNLQGSNGGYAWEDEIKRSWDLVKVDDEGDMASLVASIVEARKKRTAKKNITPYQRGIIRSLILTLDCSEAMLEKDLRPNRHAMIIQYAIDFVHEFFDQNPISQMGIIIMRNGLAQLVSQVSGNPQDHIDALKSIRKQEPKGNPSLQNALEMARGLLLPVPAHCTREVLIVFGSLSTTDPGDIHQTIDSLVSEKIRVKVLGLSAQVAICKELCKATNYGDESFYKILLDETHLKELFNEAVTPLPVNKINKGFTLVKMGFPTRIFEDTPTFCSCHSKLVYGGYFCPNCHSKVCSLPTVCPCCDLMLILSTHLARSYHHLMPLKTFAEVPTTEKFRSEDCFSCQSRFPILKNHKNGKLLTSSRYRCEDCKQEFCVDCDVFIHEILHNCPGCESKPVIT 57ADCB630B790B4F 57ADCB630B790B4F rad25_yeast General transcription and DNA repair factor IIH helicase subunit XPB YIL143C SSL2 RAD25 UVS112 LOM3 Suppressor of stem-loop mutation 2 DNA repair helicase RAD25 RNA polymerase II transcription factor B subunit SSL2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MTDVEGYQPKSKGKIFPDMGESFFSSDEDSPATDAEIDENYDDNRETSEGRGERDTGAMVTGLKKPRKKTKSSRHTAADSSMNQMDAKDKALLQDTNSDIPADFVPDSVSGMFRSHDFSYLRLRPDHASRPLWISPSDGRIILESFSPLAEQAQDFLVTIAEPISRPSHIHEYKITAYSLYAAVSVGLETDDIISVLDRLSKVPVAESIINFIKGATISYGKVKLVIKHNRYFVETTQADILQMLLNDSVIGPLRIDSDHQVQPPEDVLQQQLQQTAGKPATNVNPNDVEAVFSAVIGGDNEREEEDDDIDAVHSFEIANESVEVVKKRCQEIDYPVLEEYDFRNDHRNPDLDIDLKPSTQIRPYQEKSLSKMFGNGRARSGIIVLPCGAGKTLVGITAACTIKKSVIVLCTSSVSVMQWRQQFLQWCTLQPENCAVFTSDNKEMFQTESGLVVSTYSMVANTRNRSHDSQKVMDFLTGREWGFIILDEVHVVPAAMFRRVVSTIAAHAKLGLTATLVREDDKIGDLNFLIGPKLYEANWMELSQKGHIANVQCAEVWCPMTAEFYQEYLRETARKRMLLYIMNPTKFQACQFLIQYHERRGDKIIVFSDNVYALQEYALKMGKPFIYGSTPQQERMNILQNFQYNDQINTIFLSKVGDTSIDLPEATCLIQISSHYGSRRQEAQRLGRILRAKRRNDEGFNAFFYSLVSKDTQEMYYSTKRQAFLVDQGYAFKVITHLHGMENIPNLAYASPRERRELLQEVLLKNEEAAGIEVGDDADNSVGRGSNGHKRFKSKAVRGEGSLSGLAGGEDMAYMEYSTNKNKELKEHHPLIRKMYYKNLKK FA4013E8156FE1C5 FA4013E8156FE1C5 rad3_yeast General transcription and DNA repair factor IIH helicase subunit XPD RAD3 REM1 DNA repair helicase RAD3 RNA polymerase II transcription factor B subunit RAD3 YER171W protein protein yeast Saccharomyces cerevisiae Baker's yeast MKFYIDDLPVLFPYPKIYPEQYNYMCDIKKTLDVGGNSILEMPSGTGKTVSLLSLTIAYQMHYPEHRKIIYCSRTMSEIEKALVELENLMDYRTKELGYQEDFRGLGLTSRKNLCLHPEVSKERKGTVVDEKCRRMTNGQAKRKLEEDPEANVELCEYHENLYNIEVEDYLPKGVFSFEKLLKYCEEKTLCPYFIVRRMISLCNIIIYSYHYLLDPKIAERVSNEVSKDSIVIFDEAHNIDNVCIESLSLDLTTDALRRATRGANALDERISEVRKVDSQKLQDEYEKLVQGLHSADILTDQEEPFVETPVLPQDLLTEAIPGNIRRAEHFVSFLKRLIEYLKTRMKVLHVISETPKSFLQHLKQLTFIERKPLRFCSERLSLLVRTLEVTEVEDFTALKDIATFATLISTYEEGFLLIIEPYEIENAAVPNPIMRFTCLDASIAIKPVFERFSSVIITSGTISPLDMYPRMLNFKTVLQKSYAMTLAKKSFLPMIITKGSDQVAISSRFEIRNDPSIVRNYGSMLVEFAKITPDGMVVFFPSYLYMESIVSMWQTMGILDEVWKHKLILVETPDAQETSLALETYRKACSNGRGAILLSVARGKVSEGIDFDHQYGRTVLMIGIPFQYTESRILKARLEFMRENYRIRENDFLSFDAMRHAAQCLGRVLRGKDDYGVMVLADRRFSRKRSQLPKWIAQGLSDADLNLSTDMAISNTKQFLRTMAQPTDPKDQEGVSVWSYEDLIKHQNSRKDQGGFIENENKEGEQDEDEDEDIEMQ 978BCA01751949B4 978BCA01751949B4 rad14_yeast DNA repair protein RAD14 YM8325.02C YMR201C RAD14 protein protein yeast Saccharomyces cerevisiae Baker's yeast MTPEQKAKLEANRKLAIERLRKRGILSSDQLNRIESRNEPLKTRPLAVTSGSNRDDNAAAAVHVPNHNGQPSALANTNTNTTSLYGSGVVDGSKRDASVLDKRPTDRIRPSIRKQDYIEYDFATMQNLNGGYINPKDKLPNSDFTDDQEFESEFGSKKQKTLQDWKKEQLERKMLYENAPPPEHISKAPKCIECHINIEMDPVLHDVFKLQVCKQCSKEHPEKYALLTKTECKEDYFLTDPELNDEDLFHRLEKPNPHSGTFARMQLFVRCEVEAFAFKKWGGEEGLDEEWQRREEGKAHRREKKYEKKIKEMRLKTRAQEYTNRLREKKHGKAHIHHFSDPVDGGIDEDGYQIQRRRCTDCGLETEEIDI A90FA9CAA6F3EA52 A90FA9CAA6F3EA52 rad10_yeast DNA repair protein RAD10 YML095C RAD10 protein protein yeast Saccharomyces cerevisiae Baker's yeast MNNTDPTSFESILAGVAKLRKEKSGADTTGSQSLEIDASKLQQQEPQTSRRINSNQVINAFNQQKPEEWTDSKATDDYNRKRPFRSTRPGKTVLVNTTQKENPLLNHLKSTNWRYVSSTGINMIYYDYLVRGRSVLFLTLTYHKLYVDYISRRMQPLSRNENNILIFIVDDNNSEDTLNDITKLCMFNGFTLLLAFNFEQAAKYIEYLNL 26891AD9CA1E3E2B 26891AD9CA1E3E2B rad1_yeast DNA repair protein RAD1 YPL022W RAD1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSQLFYQGDSDDELQEELTRQTTQASQSSKIKNEDEPDDSNHLNEVENEDSKVLDDDAVLYPLIPNEPDDIETSKPNINDIRPVDIQLTLPLPFQQKVVENSLITEDALIIMGKGLGLLDIVANLLHVLATPTSINGQLKRALVLVLNAKPIDNVRIKEALEELSWFSNTGKDDDDTAVESDDELFERPFNVVTADSLSIEKRRKLYISGGILSITSRILIVDLLSGIVHPNRVTGMLVLNADSLRHNSNESFILEIYRSKNTWGFIKAFSEAPETFVMEFSPLRTKMKELRLKNVLLWPRFRVEVSSCLNATNKTSHNKVIEVKVSLTNSMSQIQFGLMECLKKCIAELSRKNPELALDWWNMENVLDINFIRSIDSVMVPNWHRISYESKQLVKDIRFLRHLLKMLVTSDAVDFFGEIQLSLDANKPSVSRKYSESPWLLVDEAQLVISYAKKRIFYKNEYTLEENPKWEQLIHILHDISHERMTNHLQGPTLVACSDNLTCLELAKVLNASNKKRGVRQVLLNKLKWYRKQREETKKLVKEVQSQDTFPENATLNVSSTFSKEQVTTKRRRTRGASQVAAVEKLRNAGTNVDMEVVFEDHKLSEEIKKGSGDDLDDGQEENAANDSKIFEIQEQENEILIDDGDAEFDNGELEYVGDLPQHITTHFNKDLWAEHCNEYEYVDRQDEILISTFKSLNDNCSLQEMMPSYIIMFEPDISFIRQIEVYKAIVKDLQPKVYFMYYGESIEEQSHLTAIKREKDAFTKLIRENANLSHHFETNEDLSHYKNLAERKLKLSKLRKSNTRNAGGQQGFHNLTQDVVIVDTREFNASLPGLLYRYGIRVIPCMLTVGDYVITPDICLERKSISDLIGSLQNNRLANQCKKMLKYYAYPTLLIEFDEGQSFSLEPFSERRNYKNKDISTVHPISSKLSQDEIQLKLAKLVLRFPTLKIIWSSSPLQTVNIILELKLGREQPDPSNAVILGTNKVRSDFNSTAKGLKDGDNESKFKRLLNVPGVSKIDYFNLRKKIKSFNKLQKLSWNEINELINDEDLTDRIYYFLRTEKEEQEQESTDENLESPGKTTDDNALHDHHNDVPEAPV 06FDA601F3F59B10 06FDA601F3F59B10 tfb1_yeast General transcription and DNA repair factor IIH subunit TFB1 TFB1 RNA polymerase II transcription factor B p73 subunit RNA polymerase II transcription factor B 73 kDa subunit RNA polymerase II transcription factor B subunit 1 D9740.3 YDR311W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSHSGAAIFEKVSGIIAINEDVSPAELTWRSTDGDKVHTVVLSTIDKLQATPASSEKMMLRLIGKVDESKKRKDNEGNEVVPKPQRHMFSFNNRTVMDNIKMTLQQIISRYKDADIYEEKRRREESAQHTETPMSSSSVTAGTPTPHLDTPQLNNGAPLINTAKLDDSLSKEKLLTNLKLQQSLLKGNKVLMKVFQETVINAGLPPSEFWSTRIPLLRAFALSTSQKVGPYNVLSTIKPVASSENKVNVNLSREKILNIFENYPIVKKAYTDNVPKNFKEPEFWARFFSSKLFRKLRGEKIMQNDRGDVIIDRYLTLDQEFDRKDDDMLLHPVKKIIDLDGNIQDDPVVRGNRPDFTMQPGVDINGNSDGTVDILKGMNRLSEKMIMALKNEYSRTNLQNKSNITNDEEDEDNDERNELKIDDLNESYKTNYAIIHLKRNAHEKTTDNDAKSSADSIKNADLKVSNQQMLQQLSLVMDNLINKLDLNQVVPNNEVSNKINKRVITAIKINAKQAKHNNVNSALGSFVDNTSQANELEVKSTLPIDLLESCRMLHTTCCEFLKHFYIHFQSGEQKQASTVKKLYNHLKDCIEKLNELFQDVLNGDGESMSNTCTAYLKPVLNSITLATHKYDEYFNEYNNNSN DAE369FE90A224F8 DAE369FE90A224F8 tfb1-rad25-1 1 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association The Rad3, Rad25, and TFB1 subunits of TFIIH in the 100 mM imidazole wash are marked in B. Fig. 1 tfb1-ssl1-1 2 5 unspecified role unspecified role neutral component neutral component 11 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component physical association physical association Fig. 2B rad1-rad10-1 2 10 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component physical association physical association It was of particular interest, however, that Rad14 protein (Mr 43,000) co-eluted with the Rad1-Rad10 complex (combined Mr 150,700) from the S-300 molecular siz-ing step (Fig. 2B), Fig. 2B rad1-rad10-2 3 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait association association When the S-300 fractions containing the peak of the Rad1, Rad10, and Rad14 proteins were subjected to immunoprecipitation with protein A-agarose beads bearing anti-Rad1, anti-Rad10, and anti-Rad14 antibodies, we observed the co-precipitation of the three proteins (Fig. 3). Fig. 3 rad1-rad10-3 3 8 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait association association When the S-300 fractions containing the peak of the Rad1, Rad10, and Rad14 proteins were subjected to immunoprecipitation with protein A-agarose beads bearing anti-Rad1, anti-Rad10, and anti-Rad14 antibodies, we observed the co-precipitation of the three proteins (Fig. 3). Fig. 3 rad1-rad10-4 3 10 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait association association When the S-300 fractions containing the peak of the Rad1, Rad10, and Rad14 proteins were subjected to immunoprecipitation with protein A-agarose beads bearing anti-Rad1, anti-Rad10, and anti-Rad14 antibodies, we observed the co-precipitation of the three proteins (Fig. 3). Fig. 3 rad1-rad14-1 4 10 unspecified role unspecified role bait bait yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 8 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro association association As shown in Fig. 5, Rad1 protein alone interacts with Rad14 protein, Fig. 5 Levels of RAD14 coprecipitated with RAD1 increases when RAD10 is present, suggesting that a Rad1-Rad10 complex recruits Rad14. rad1-rad14-2 4 8 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 10 unspecified role unspecified role ancillary ancillary yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 9 unspecified role unspecified role bait bait yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast association association The Rad1-Rad10 complex is the molecular entity for Rad14 recruitment. Fig. 5