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/ Retrotranslocation of a Misfolded Luminal ER Protein by the Ubiquitin-Ligase Hrd1p. Retrotranslocation of a Misfolded Luminal ER Protein by the Ubiquitin-Ligase Hrd1p. Cell (0092-8674) 2010 imex curation Carvalho P., Stanley AM., Rapoport TA. Crosslinking agents 0.2 mM disuccinimidyl suberate (DSS) or 0.2 mM ethylene glycol bis[succinimidylsuccinate] (EGS) - proteins were then immunoprecipitated. pedro.carvalho@crg.es, tom_rapoport@hms.harvard.edu 06-April-2011: Contacted by JYOTI. mgt Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Carvalho P., Stanley AM., Rapoport TA. Crosslinking agents 0.2 mM disuccinimidyl suberate (DSS) or 0.2 mM ethylene glycol bis[succinimidylsuccinate] (EGS) - proteins were then immunoprecipitated. pedro.carvalho@crg.es, tom_rapoport@hms.harvard.edu Accepted 2010-DEC-14 by MGT 06-April-2011: Contacted by JYOTI. Cell (0092-8674) 2010 mgt Only protein-protein interactions imex curation Retrotranslocation of a Misfolded Luminal ER Protein by the Ubiquitin-Ligase Hrd1p. Retrotranslocation of a Misfolded Luminal ER Protein by the Ubiquitin-Ligase Hrd1p. Cell (0092-8674) 2010 imex curation Carvalho P., Stanley AM., Rapoport TA. Crosslinking agents 0.2 mM disuccinimidyl suberate (DSS) or 0.2 mM ethylene glycol bis[succinimidylsuccinate] (EGS) - proteins were then immunoprecipitated. pedro.carvalho@crg.es, tom_rapoport@hms.harvard.edu 06-April-2011: Contacted by JYOTI. mgt Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast tap tandem affinity purification TAP TAP tap ms participant Identification by mass spectrometry ms participant Carvalho P., Stanley AM., Rapoport TA. Crosslinking agents 0.2 mM disuccinimidyl suberate (DSS) or 0.2 mM ethylene glycol bis[succinimidylsuccinate] (EGS) - proteins were then immunoprecipitated. pedro.carvalho@crg.es, tom_rapoport@hms.harvard.edu Accepted 2010-DEC-14 by MGT 06-April-2011: Contacted by JYOTI. Cell (0092-8674) 2010 mgt Only protein-protein interactions imex curation Retrotranslocation of a Misfolded Luminal ER Protein by the Ubiquitin-Ligase Hrd1p. Retrotranslocation of a Misfolded Luminal ER Protein by the Ubiquitin-Ligase Hrd1p. Cell (0092-8674) 2010 imex curation Carvalho P., Stanley AM., Rapoport TA. Crosslinking agents 0.2 mM disuccinimidyl suberate (DSS) or 0.2 mM ethylene glycol bis[succinimidylsuccinate] (EGS) - proteins were then immunoprecipitated. pedro.carvalho@crg.es, tom_rapoport@hms.harvard.edu 06-April-2011: Contacted by JYOTI. mgt Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast crosslink cross-linking study crosslink anti tag western anti tag western blot anti tag western Carvalho P., Stanley AM., Rapoport TA. Crosslinking agents 0.2 mM disuccinimidyl suberate (DSS) or 0.2 mM ethylene glycol bis[succinimidylsuccinate] (EGS) - proteins were then immunoprecipitated. pedro.carvalho@crg.es, tom_rapoport@hms.harvard.edu Accepted 2010-DEC-14 by MGT 06-April-2011: Contacted by JYOTI. Cell (0092-8674) 2010 mgt Only protein-protein interactions imex curation Retrotranslocation of a Misfolded Luminal ER Protein by the Ubiquitin-Ligase Hrd1p. Retrotranslocation of a Misfolded Luminal ER Protein by the Ubiquitin-Ligase Hrd1p. Cell (0092-8674) 2010 imex curation Carvalho P., Stanley AM., Rapoport TA. Crosslinking agents 0.2 mM disuccinimidyl suberate (DSS) or 0.2 mM ethylene glycol bis[succinimidylsuccinate] (EGS) - proteins were then immunoprecipitated. pedro.carvalho@crg.es, tom_rapoport@hms.harvard.edu 06-April-2011: Contacted by JYOTI. mgt Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast crosslink cross-linking study crosslink anti tag western anti tag western blot anti tag western Carvalho P., Stanley AM., Rapoport TA. Crosslinking agents 0.2 mM disuccinimidyl suberate (DSS) or 0.2 mM ethylene glycol bis[succinimidylsuccinate] (EGS) - proteins were then immunoprecipitated. pedro.carvalho@crg.es, tom_rapoport@hms.harvard.edu Accepted 2010-DEC-14 by MGT 06-April-2011: Contacted by JYOTI. Cell (0092-8674) 2010 mgt Only protein-protein interactions imex curation hrd1_yeast ERAD-associated E3 ubiquitin-protein ligase HRD1 HMG-CoA reductase degradation protein 1 HRD1 DER3 RING-type E3 ubiquitin transferase HRD1 YOL013C protein protein yeast Saccharomyces cerevisiae Baker's yeast MVPENRRKQLAIFVVVTYLLTFYCVYSATKTSVSFLQVTLKLNEGFNLMVLSIFILLNSTLLWQLLTKLLFGELRLIEHEHIFERLPFTIINTLFMSSLFHERYFFTVAFFGLLLLYLKVFHWILKDRLEALLQSINDSTTMKTLIFSRFSFNLVLLAVVDYQIITRCISSIYTNQKSDIESTSLYLIQVMEFTMLLIDLLNLFLQTCLNFWEFYRSQQSLSNENNHIVHGDPTDENTVESDQSQPVLNDDDDDDDDDRQFTGLEGKFMYEKAIDVFTRFLKTALHLSMLIPFRMPMMLLKDVVWDILALYQSGTSLWKIWRNNKQLDDTLVTVTVEQLQNSANDDNICIICMDELIHSPNQQTWKNKNKKPKRLPCGHILHLSCLKNWMERSQTCPICRLPVFDEKGNVVQTTFTSNSDITTQTTVTDSTGIATDQQGFANEVDLLPTRTTSPDIRIVPTQNIDTLAMRTRSTSTPSPTWYTFPLHKTGDNSVGSSRSAYEFLITNSDEKENGIPVKLTIENHEVNSLHGDGGEQIAKKIVIPDKFIQHI CAA6341E7A94DB0B usa1_yeast U1 SNP1-associating protein 1 USA1 YML029W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSEYLAQTPCKFTIWSSEIDLIRTNLLVNAHPLSTVGRLLQYIHYQIYKQLRAIYQPEEQCTNSEIPHTPLNSINTYFLSYEGRELSATCLLKDITSSSHPDSNHFIRLQLEKRTSPSGSAFDLEYDMEGEFNSMNIQFEINTLSSQRIFNSMEPNLPIGTTLARLEKLALERIKDFEKSAGNLCGIKEDHSVSDLQGFIIKGKQTPMFLNYGSDSDYYKDLNLVDLIGIDFAPAHNSFFTFLFKMNHEQNSHIANDEERFVLEFISDATLSITQMNVKPDTTVKQVKDFICSVYTHSLNLRRNDIKLIYKGQLLHENNFAGNSSKISEYIKEPHEVKVHVQINQEYTESGPGFWNEVFNNPNIFQFMPPDTRSQSPVSFAPTQGRSPAAIRGEERGIPYVTESGNDIVPTDELYRKCIINGDEVVFIPVSELNPQSSYLSVIKGDYGEIKIPISSNDYRINGDNILLSPSAIEQLESALNFKIERPRDSTLLHPSGEHVRAADNTSSANDNNTVENDESAWNRRVVRPLRNSFPLLLVLIRTFYLIGYNSLVPFFIILEFGSFLPWKYIILLSLLFIFRTVWNTQEVWNLWRDYLHLNEIDEVKFSQIKEFINSNSLTLNFYKKCKDTQSAIDLLMIPNLHEQRLSVYSKYDIEYDTNTPDVGQLNLLFIKVLSGEIPKDALDELFKEFFELYETTRNMNTLYPQDSLNELLLMIWKESQKKDINTLPKYRRWFQTLCSQIAEHNVLDVVLRYIIPDPVNDRVITAVIKNFVLFWVTLLPYVKEKLDDIVAQRARDREQPAPSAQQQENEDEALIIPDEEEPTATGAQPHLYIPDED 9B93ECA6C5421FD6 9B93ECA6C5421FD6 cbpy_yeast Carboxypeptidase Y PRC1 YMR297W Carboxypeptidase YSCY protein protein yeast Saccharomyces cerevisiae Baker's yeast MKAFTSLLCGLGLSTTLAKAISLQRPLGLDKDVLLQAAEKFGLDLDLDHLLKELDSNVLDAWAQIEHLYPNQVMSLETSTKPKFPEAIKTKKDWDFVVKNDAIENYQLRVNKIKDPKILGIDPNVTQYTGYLDVEDEDKHFFFWTFESRNDPAKDPVILWLNGGPGCSSLTGLFFELGPSSIGPDLKPIGNPYSWNSNATVIFLDQPVNVGFSYSGSSGVSNTVAAGKDVYNFLELFFDQFPEYVNKGQDFHIAGESYAGHYIPVFASEILSHKDRNFNLTSVLIGNGLTDPLTQYNYYEPMACGEGGEPSVLPSEECSAMEDSLERCLGLIESCYDSQSVWSCVPATIYCNNAQLAPYQRTGRNVYDIRKDCEGGNLCYPTLQDIDDYLNQDYVKEAVGAEVDHYESCNFDINRNFLFAGDWMKPYHTAVTDLLNQDLPILVYAGDKDFICNWLGNKAWTDVLPWKYDEEFASQKVRNWTASITDEVAGEVKSYKHFTYLRVFNGGHMVPFDVPENALSMVNEWIHGGFSL 7227F3489CBDD952 7227F3489CBDD952 der1_yeast Degradation in the endoplasmic reticulum protein 1 DER1 YBR1413 YBR201W protein protein yeast Saccharomyces cerevisiae Baker's yeast MDAVILNLLGDIPLVTRLWTIGCLVLSGLTSLRIVDPGKVVYSYDLVFKKGQYGRLLYSIFDYGAFNWISMINIFVSANHLSTLENSFNLRRKFCWIIFLLLVILVKMTSIEQPAASLGVLLHENLVYYELKKNGNQMNVRFFGAIDVSPSIFPIYMNAVMYFVYKRSWLEIAMNFMPGHVIYYMDDIIGKIYGIDLCKSPYDWFRNTETP 41510999E92C691D os9_yeast Protein OS-9 homolog YOS9 D4222 YDR057W protein protein yeast Saccharomyces cerevisiae Baker's yeast MQAKIIYALSAISALIPLGSSLLAPIEDPIVSNKYLISYIDEDDWSDRILQNQSVMNSGYIVNMGDDLECFIQNASTQLNDVLEDSNEHSNSEKTALLTKTLNQGVKTIFDKLNERCIFYQAGFWIYEYCPGIEFVQFHGRVNTKTGEIVNRDESLVYRLGKPKANVEEREFELLYDDVGYYISEIIGSGDICDVTGAERMVEIQYVCGGSNSGPSTIQWVRETKICVYEAQVTIPELCNLELLAKNEDQKNASPILCRMPAKSKIGSNSIDLITKYEPIFLGSGIYFLRPFNTDERDKLMVTDNAMSNWDEITETYYQKFGNAINKMLSLRLVSLPNGHILQPGDSCVWLAEVVDMKDRFQTTLSLNILNSQRAEIFFNKTFTFNEDNGNFLSYKIGDHGESTELGQITHSNKADINTAEIRSDEYLINTDNELFLRISKEIAEVKELLNEIVSPHEMEVIFENMRNQPNNDFELALMNKLKSSLNDDNKVEQINNARMDDDESTSHTTRDIGEAGSQTTGNTESEVTNVAAGVFIEHDEL BF353A0278703BE8 hrd3_yeast ERAD-associated E3 ubiquitin-protein ligase component HRD3 HMG-CoA reductase degradation protein 3 HRD3 YLR207W protein protein yeast Saccharomyces cerevisiae Baker's yeast MITLLLYLCVICNAIVLIRADSIADPWPEARHLLNTIAKSRDPMKEAAMEPNADEFVGFYVPMDYSPRNEEKNYQSIWQNEITDSQRHIYELLVQSSEQFNNSEATYTLSQIHLWSQYNFPHNMTLAHKYLEKFNDLTHFTNHSAIFDLAVMYATGGCASGNDQTVIPQDSAKALLYYQRAAQLGNLKAKQVLAYKYYSGFNVPRNFHKSLVLYRDIAEQLRKSYSRDEWDIVFPYWESYNVRISDFESGLLGKGLNSVPSSTVRKRTTRPDIGSPFIAQVNGVQMTLQIEPMGRFAFNGNDGNINGDEDDEDASERRIIRIYYAALNDYKGTYSQSRNCERAKNLLELTYKEFQPHVDNLDPLQVFYYVRCLQLLGHMYFTGEGSSKPNIHMAEEILTTSLEISRRAQGPIGRACIDLGLINQYITNNISQAISYYMKAMKTQANNGIVEFQLSKLATSFPEEKIGDPFNLMETAYLNGFIPAIYEFAVMIESGMNSKSSVENTAYLFKTFVDKNEAIMAPKLRTAFAALINDRSEVALWAYSQLAEQGYETAQVSAAYLMYQLPYEFEDPPRTTDQRKTLAISYYTRAFKQGNIDAGVVAGDIYFQMQNYSKAMALYQGAALKYSIQAIWNLGYMHEHGLGVNRDFHLAKRYYDQVSEHDHRFYLASKLSVLKLHLKSWLTWITREKVNYWKPSSPLNPNEDTQHSKTSWYKQLTKILQRMRHKEDSDKAAEDSHKHRTVVQNGANHRGDDQEEASEILGFQMEDLVTMGCILGIFLLSILMSTLAARRGWNVRFNGAQLNANGNRQQEQQQQQQAQGPPGWDFNVQIFAI E7DB29FBA16D9BFD ubx2_yeast UBX domain-containing protein 2 SEL1 YM9571.05 YML013W UBX2 Secretion lowering protein 1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MPVVNHEDSEFHLSHTEEDKLNEFQVITNFPPEDLPDVVRLLRNHGWQLEPALSRYFDGEWKGEPDQMGEPTQTSTPMAETLVPPALGPRPLLFTASLPVVRPLPANFRNDFRTIGLNGRSNTVWSMFESFSYDGNPFLFILLLIPRIINRLSATIFTFFCTLLSLHSISGGGNSGKPKISKVPKAPTRETHIPLAEILGDTKDKDAFCELKSFKPDISFNEALRIAKEEFKFMLLILVGDTYDTDTDTVDVNSKLLLEKILLNKKTLQYLRKIDNDLIIYLKCVHELEPWLVARQLGVRNTPEIFLIANVANKASHSETLPSQRLSILGKLKVNSLNRFLQSLTNVVEKYTPELVVNKTEMHELRMSREIKKLQEDAYKKSLEMDRIKAIEKEKSLKHAQDLKLNSTARQLKWLKACIDEIQPFETTGKQATLQFRTSSGKRFVKKFPSMTTLYQIYQSIGCHIYLAVYSSDPAEWSNALQDKIRQLSADDDMLCFKEGQLETATATTIEELGHIINNELTSFDLERGKLEFDFELVSPFPKYTVHPNEHMSVDQVPQLWPNGSLLVEALDEEDEEDEENEEQ DA4DDD3D624C17B8 DA4DDD3D624C17B8 cdc48_yeast Cell division control protein 48 CDC48 Cell division cycle protein 48 Transitional endoplasmic reticulum ATPase homolog YDL126C protein protein yeast Saccharomyces cerevisiae Baker's yeast MGEEHKPLLDASGVDPREEDKTATAILRRKKKDNMLLVDDAINDDNSVIAINSNTMDKLELFRGDTVLVKGKKRKDTVLIVLIDDELEDGACRINRVVRNNLRIRLGDLVTIHPCPDIKYATRISVLPIADTIEGITGNLFDVFLKPYFVEAYRPVRKGDHFVVRGGMRQVEFKVVDVEPEEYAVVAQDTIIHWEGEPINREDEENNMNEVGYDDIGGCRKQMAQIREMVELPLRHPQLFKAIGIKPPRGVLMYGPPGTGKTLMARAVANETGAFFFLINGPEVMSKMAGESESNLRKAFEEAEKNAPAIIFIDEIDSIAPKRDKTNGEVERRVVSQLLTLMDGMKARSNVVVIAATNRPNSIDPALRRFGRFDREVDIGIPDATGRLEVLRIHTKNMKLADDVDLEALAAETHGYVGADIASLCSEAAMQQIREKMDLIDLDEDEIDAEVLDSLGVTMDNFRFALGNSNPSALRETVVESVNVTWDDVGGLDEIKEELKETVEYPVLHPDQYTKFGLSPSKGVLFYGPPGTGKTLLAKAVATEVSANFISVKGPELLSMWYGESESNIRDIFDKARAAAPTVVFLDELDSIAKARGGSLGDAGGASDRVVNQLLTEMDGMNAKKNVFVIGATNRPDQIDPAILRPGRLDQLIYVPLPDENARLSILNAQLRKTPLEPGLELTAIAKATQGFSGADLLYIVQRAAKYAIKDSIEAHRQHEAEKEVKVEGEDVEMTDEGAKAEQEPEVDPVPYITKEHFAEAMKTAKRSVSDAELRRYEAYSQQMKASRGQFSNFNFNDAPLGTTATDNANSNNSAPSGAGAAFGSNAEEDDDLYS 02ADDB9A227614D8 02ADDB9A227614D8 hrd1-1 4 5 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Interaction greatly reduced in usa-/- cells When proteins are crosslinked, oligomers form. Fig. 2A hrd1-2 1 5 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Interaction greatly reduced in usa-/- cells. Hrd1 (cys399ser) oligomerized even in the absence of Usa1. Figs. 2A, 2D, Supp. Fig. 3A hrd1-usa1-1 1 6 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 5 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain association association Fig. 2B, Supp. Fig 3A Q03714 deletion 437-490 abolished both the binding of Usa1 to Hrd1 and the oligomerization of Hrd1, the region from 371 to 418 also appears to be important for Hrd1p oligomerization. usa1-1 1 6 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 6 unspecified role unspecified role prey prey 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 physical association physical association Fig. 2C hrd1-usa1-4g 4 5 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig. 2A The arrowhead indicates the position of a crosslinked species containing both Hrd1p and Usa1p. prc1-hrd1-1 3 5 unspecified role unspecified role prey prey c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Q08109:p.Cys399Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 20:13:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys399ser' Tue Jun 30 18:40:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys399Ser' 7 unspecified role unspecified role bait bait c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminal dihydrofolate reductase fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 52 47 physical association physical association Figs. 4A, 4B, 5A, 5B, 5C, 5D, 6A, 6B, Supp. Fig. 7A Mutation of a glycosylation site (position not given) of Hrd1 reduced binding to PRC1. Interaction was also reduced in a OS9-/- cell line and absent in cells expressing a fragment (residues 357 833) of the luminal domain of Hrd3p. Binding was also reduced in Der1-/- and Usa1-/- cells but retained in Usa1 deletion mutant clss provided region 437 490 of Usa1 was present. prc1-hrd3-1 3 10 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 c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 7 unspecified role unspecified role bait bait c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminal dihydrofolate reductase fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Fig. 4C prc1-der1-1 3 7 unspecified role unspecified role bait bait c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminal dihydrofolate reductase fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role prey prey c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Fig. 4D hrd1-hrd3-1 2 11 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 5 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 certain certain sequence position certain certain certain sequence position certain region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Supp. Fig. 5C prc1-hrd1-2 3 5 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 c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 7 unspecified role unspecified role bait bait c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminal dihydrofolate reductase fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 79 84 physical association physical association Figs. 4F Mini-Hrd1p no longer gave photocrosslinks to sCPY*-DHFR-HA (Figure 4F)