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/ A chaperone cascade sorts proteins for posttranslational membrane insertion into the endoplasmic reticulum. A chaperone cascade sorts proteins for posttranslational membrane insertion into the endoplasmic reticulum. Molecular cell 2010 imex curation Wang F., Brown EC., Mak G., Zhuang J., Denic V., Wang F., Brown EC., Mak G., Zhuang J., Denic V. vdenic@mcb.harvard.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Wang F., Brown EC., Mak G., Zhuang J., Denic V., Wang F., Brown EC., Mak G., Zhuang J., Denic V. I have only dealt with the competition expts as comments because 1. I'm not sure which constructs they are using (I suspect His tagged/Ecoli expressed), 2. I don't know whether the elution protein is binding to the tagged protein or the prey, 3. we haven't previously regarded elution proteins/peptides as prey, we've just ignored them and 4. when they start throwing 3 or 4 proteins in, it just gets far too complicated, so I decided consistent comments across the board was the simplest solution. vdenic@mcb.harvard.edu Accepted 2018-JUL-13 AT 08:18 BST AT 08:18 BST by PPORRAS Molecular cell 2010 Only protein-protein interactions imex curation imex curation Removed - though i should point out Livia was having me add this when I did these so we have entries out there with it on. A chaperone cascade sorts proteins for posttranslational membrane insertion into the endoplasmic reticulum. A chaperone cascade sorts proteins for posttranslational membrane insertion into the endoplasmic reticulum. Molecular cell 2010 imex curation Wang F., Brown EC., Mak G., Zhuang J., Denic V., Wang F., Brown EC., Mak G., Zhuang J., Denic V. vdenic@mcb.harvard.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip weight by sybr molecular weight estimation by sybr staining safe DNA gel stain safe DNA gel stain weight by sybr Wang F., Brown EC., Mak G., Zhuang J., Denic V., Wang F., Brown EC., Mak G., Zhuang J., Denic V. vdenic@mcb.harvard.edu Accepted 2018-JUL-13 AT 08:18 BST AT 08:18 BST by PPORRAS Molecular cell 2010 Only protein-protein interactions imex curation imex curation Removed - though i should point out Livia was having me add this when I did these so we have entries out there with it on. A chaperone cascade sorts proteins for posttranslational membrane insertion into the endoplasmic reticulum. A chaperone cascade sorts proteins for posttranslational membrane insertion into the endoplasmic reticulum. Molecular cell 2010 imex curation Wang F., Brown EC., Mak G., Zhuang J., Denic V., Wang F., Brown EC., Mak G., Zhuang J., Denic V. vdenic@mcb.harvard.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation ecoli Escherichia coli (strain K12) anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Wang F., Brown EC., Mak G., Zhuang J., Denic V., Wang F., Brown EC., Mak G., Zhuang J., Denic V. I would normally just add the SUMO as a PTM, but in this case, it sounds more like a coexpression. vdenic@mcb.harvard.edu Accepted 2018-JUL-13 AT 08:18 BST AT 08:18 BST by PPORRAS Molecular cell 2010 Only protein-protein interactions imex curation imex curation Removed - though i should point out Livia was having me add this when I did these so we have entries out there with it on. A chaperone cascade sorts proteins for posttranslational membrane insertion into the endoplasmic reticulum. A chaperone cascade sorts proteins for posttranslational membrane insertion into the endoplasmic reticulum. Molecular cell 2010 imex curation Wang F., Brown EC., Mak G., Zhuang J., Denic V., Wang F., Brown EC., Mak G., Zhuang J., Denic V. vdenic@mcb.harvard.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip sequence tag sequence tag identification sequence tag Wang F., Brown EC., Mak G., Zhuang J., Denic V., Wang F., Brown EC., Mak G., Zhuang J., Denic V. vdenic@mcb.harvard.edu Accepted 2018-JUL-13 AT 08:18 BST AT 08:18 BST by PPORRAS Molecular cell 2010 Only protein-protein interactions imex curation imex curation Removed - though i should point out Livia was having me add this when I did these so we have entries out there with it on. A chaperone cascade sorts proteins for posttranslational membrane insertion into the endoplasmic reticulum. A chaperone cascade sorts proteins for posttranslational membrane insertion into the endoplasmic reticulum. Molecular cell 2010 imex curation Wang F., Brown EC., Mak G., Zhuang J., Denic V., Wang F., Brown EC., Mak G., Zhuang J., Denic V. vdenic@mcb.harvard.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Wang F., Brown EC., Mak G., Zhuang J., Denic V., Wang F., Brown EC., Mak G., Zhuang J., Denic V. I would normally just add the SUMO as a PTM, but in this case, it sounds more like a coexpression. vdenic@mcb.harvard.edu Accepted 2018-JUL-13 AT 08:18 BST AT 08:18 BST by PPORRAS Molecular cell 2010 Only protein-protein interactions imex curation imex curation Removed - though i should point out Livia was having me add this when I did these so we have entries out there with it on. A chaperone cascade sorts proteins for posttranslational membrane insertion into the endoplasmic reticulum. A chaperone cascade sorts proteins for posttranslational membrane insertion into the endoplasmic reticulum. Molecular cell 2010 imex curation Wang F., Brown EC., Mak G., Zhuang J., Denic V., Wang F., Brown EC., Mak G., Zhuang J., Denic V. vdenic@mcb.harvard.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Wang F., Brown EC., Mak G., Zhuang J., Denic V., Wang F., Brown EC., Mak G., Zhuang J., Denic V. I have only dealt with the competition expts as comments because 1. I'm not sure which constructs they are using (I suspect His tagged/Ecoli expressed), 2. I don't know whether the elution protein is binding to the tagged protein or the prey, 3. we haven't previously regarded elution proteins/peptides as prey, we've just ignored them and 4. when they start throwing 3 or 4 proteins in, it just gets far too complicated, so I decided consistent comments across the board was the simplest solution. vdenic@mcb.harvard.edu Accepted 2018-JUL-13 AT 08:18 BST AT 08:18 BST by PPORRAS Molecular cell 2010 Only protein-protein interactions imex curation imex curation Removed - though i should point out Livia was having me add this when I did these so we have entries out there with it on. sec22_yeast Protein transport protein SEC22 SEC22 SLY2 YLR268W L8479.3 TSL26 Suppressor of loss of YPT1 protein 2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MIKSTLIYREDGLPLCTSVDNENDPSLFEQKQKVKIVVSRLTPQSATEATLESGSFEIHYLKKSMVYYFVICESGYPRNLAFSYLNDIAQEFEHSFANEYPKPTVRPYQFVNFDNFLQMTKKSYSDKKVQDNLDQLNQELVGVKQIMSKNIEDLLYRGDSLDKMSDMSSSLKETSKRYRKSAQKINFDLLISQYAPIVIVAFFFVFLFWWIFLK 9FA3E34EAAFB9536 9FA3E34EAAFB9536 sis1_yeast Protein SIS1 YNL007C SIS1 N2879 protein protein yeast Saccharomyces cerevisiae Baker's yeast MVKETKLYDLLGVSPSANEQELKKGYRKAALKYHPDKPTGDTEKFKEISEAFEILNDPQKREIYDQYGLEAARSGGPSFGPGGPGGAGGAGGFPGGAGGFSGGHAFSNEDAFNIFSQFFGGSSPFGGADDSGFSFSSYPSGGGAGMGGMPGGMGGMHGGMGGMPGGFRSASSSPTYPEEETVQVNLPVSLEDLFVGKKKSFKIGRKGPHGASEKTQIDIQLKPGWKAGTKITYKNQGDYNPQTGRRKTLQFVIQEKSHPNFKRDGDDLIYTLPLSFKESLLGFSKTIQTIDGRTLPLSRVQPVQPSQTSTYPGQGMPTPKNPSQRGNLIVKYKVDYPISLNDAQKRAIDENF 642D3709C0FD5682 642D3709C0FD5682 yby7_yeast UPF0303 protein YBR137W YBR1013 YBR137W protein protein yeast Saccharomyces cerevisiae Baker's yeast MVVLDKKLLERLTSRKVPLEELEDMEKRCFLSTFTYQDAFDLGTYIRNAVKENFPEKPVAIDISLPNGHCLFRTVTYGGSALDNDFWIQRKKKTALRFGHSSFYMGCKKGDKTPEEKFFVDSKEYAFHGGAVLIQSERSDYPYACLTISGLKQEEDHLMAVSSLIAFANESLEEDLNLD 8CBA454492E29273 8CBA454492E29273 hs104_yeast Heat shock protein 104 HSP104 YLL026W L0948 Protein aggregation-remodeling factor HSP104 protein protein yeast Saccharomyces cerevisiae Baker's yeast MNDQTQFTERALTILTLAQKLASDHQHPQLQPIHILAAFIETPEDGSVPYLQNLIEKGRYDYDLFKKVVNRNLVRIPQQQPAPAEITPSYALGKVLQDAAKIQKQQKDSFIAQDHILFALFNDSSIQQIFKEAQVDIEAIKQQALELRGNTRIDSRGADTNTPLEYLSKYAIDMTEQARQGKLDPVIGREEEIRSTIRVLARRIKSNPCLIGEPGIGKTAIIEGVAQRIIDDDVPTILQGAKLFSLDLAALTAGAKYKGDFEERFKGVLKEIEESKTLIVLFIDEIHMLMGNGKDDAANILKPALSRGQLKVIGATTNNEYRSIVEKDGAFERRFQKIEVAEPSVRQTVAILRGLQPKYEIHHGVRILDSALVTAAQLAKRYLPYRRLPDSALDLVDISCAGVAVARDSKPEELDSKERQLQLIQVEIKALERDEDADSTTKDRLKLARQKEASLQEELEPLRQRYNEEKHGHEELTQAKKKLDELENKALDAERRYDTATAADLRYFAIPDIKKQIEKLEDQVAEEERRAGANSMIQNVVDSDTISETAARLTGIPVKKLSESENEKLIHMERDLSSEVVGQMDAIKAVSNAVRLSRSGLANPRQPASFLFLGLSGSGKTELAKKVAGFLFNDEDMMIRVDCSELSEKYAVSKLLGTTAGYVGYDEGGFLTNQLQYKPYSVLLFDEVEKAHPDVLTVMLQMLDDGRITSGQGKTIDCSNCIVIMTSNLGAEFINSQQGSKIQESTKNLVMGAVRQHFRPEFLNRISSIVIFNKLSRKAIHKIVDIRLKEIEERFEQNDKHYKLNLTQEAKDFLAKYGYSDDMGARPLNRLIQNEILNKLALRILKNEIKDKETVNVVLKKGKSRDENVPEEAEECLEVLPNHEATIGADTLGDDDNEDSMEIDDDLD 4AD0E7E3AF98E318 4AD0E7E3AF98E318 get4_yeast Golgi to ER traffic protein 4 GET4 O3580 YOR164C Guided entry of tail-anchored proteins 4 protein protein yeast Saccharomyces cerevisiae Baker's yeast MVPAESNAVQAKLAKTLQRFENKIKAGDYYEAHQTLRTIANRYVRSKSYEHAIELISQGALSFLKAKQGGSGTDLIFYLLEVYDLAEVKVDDISVARLVRLIAELDPSEPNLKDVITGMNNWSIKFSEYKFGDPYLHNTIGSKLLEGDFVYEAERYFMLGTHDSMIKYVDLLWDWLCQVDDIEDSTVAEFFSRLVFNYLFISNISFAHESKDIFLERFIEKFHPKYEKIDKNGYEIVFFEDYSDLNFLQLLLITCQTKDKSYFLNLKNHYLDFSQAYKSELEFLGQEYFNIVAPKQTNFLQDMMSGFLGGSK 98AFFDB4EA51B40F 98AFFDB4EA51B40F sgt2_yeast Small glutamine-rich tetratricopeptide repeat-containing protein 2 Viral protein U-binding protein SGT/UBP SGT2 UNF346 YOR007C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSASKEEIAALIVNYFSSIVEKKEISEDGADSLNVAMDCISEAFGFEREAVSGILGKSEFKGQHLADILNSASRVPESNKKDDAENVEINIPEDDAETKAKAEDLKMQGNKAMANKDYELAINKYTEAIKVLPTNAIYYANRAAAHSSLKEYDQAVKDAESAISIDPSYFRGYSRLGFAKYAQGKPEEALEAYKKVLDIEGDNATEAMKRDYESAKKKVEQSLNLEKTVPEQSRDADVDASQGASAGGLPDLGSLLGGGLGGLMNNPQLMQAAQKMMSNPGAMQNIQKMMQDPSIRQMAEGFASGGGTPNLSDLMNNPALRNMAGNLFGGAGAQSTDETPDNENKQ 33539B74F742BD3C 33539B74F742BD3C fis1_yeast Mitochondrial fission 1 protein Mitochondrial division protein 2 FIS1 MDV2 YIL065C protein protein yeast Saccharomyces cerevisiae Baker's yeast MTKVDFWPTLKDAYEPLYPQQLEILRQQVVSEGGPTATIQSRFNYAWGLIKSTDVNDERLGVKILTDIYKEAESRRRECLYYLTIGCYKLGEYSMAKRYVDTLFEHERNNKQVGALKSMVEDKIQKETLKGVVVAGGVLAGAVAVASFFLRNKRR BD62D19524770536 BD62D19524770536 smt3_yeast Ubiquitin-like protein SMT3 SMT3 D9719.15 YDR510W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGGATY A2790DE7F315E1A7 A2790DE7F315E1A7 get3_yeast ATPase GET3 YDL100C GET3 ARR4 D2371 Arsenical pump-driving ATPase Arsenite-stimulated ATPase Guided entry of tail-anchored proteins 3 Golgi to ER traffic protein 3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MDLTVEPNLHSLITSTTHKWIFVGGKGGVGKTTSSCSIAIQMALSQPNKQFLLISTDPAHNLSDAFGEKFGKDARKVTGMNNLSCMEIDPSAALKDMNDMAVSRANNNGSDGQGDDLGSLLQGGALADLTGSIPGIDEALSFMEVMKHIKRQEQGEGETFDTVIFDTAPTGHTLRFLQLPNTLSKLLEKFGEITNKLGPMLNSFMGAGNVDISGKLNELKANVETIRQQFTDPDLTTFVCVCISEFLSLYETERLIQELISYDMDVNSIIVNQLLFAENDQEHNCKRCQARWKMQKKYLDQIDELYEDFHVVKMPLCAGEIRGLNNLTKFSQFLNKEYNPITDGKVIYELEDKE F795C359B5A4A461 F795C359B5A4A461 mdy2_yeast Ubiquitin-like protein MDY2 MDY2 TMA24 Mating-deficient protein 2 Translation machinery-associated protein 24 GET5 Golgi to ER traffic protein 5 YOL111C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSTSASGPEHEFVSKFLTLATLTEPKLPKSYTKPLKDVTNLGVPLPTLKYKYKQNRAKKLKLHQDQQGQDNAAVHLTLKKIQAPKFSIEHDFSPSDTILQIKQHLISEEKASHISEIKLLLKGKVLHDNLFLSDLKVTPANSTITVMIKPNPTISKEPEAEKSTNSPAPAPPQELTVPWDDIEALLKNNFENDQAAVRQVMERLQKGWSLAK CE6FC4282E0E472C CE6FC4282E0E472C mdy2-hsp104-1 4 16 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 11 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 12 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level association association Interaction observed in GET3-/- cells. Additional chaperones, only identified as HSP70s, were also seen to have bound. Fig. 2A, Supp Fig 2A get4-hsp104-1 4 16 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 11 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 12 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level association association Interaction observed in GET3-/- cells. Additional chaperones, only identified as HSP70s, were also seen to have bound. Fig. 2A, Supp. Fig. 2A mdy2-sgt2-1 4 16 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 12 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level association association Interaction observed in GET4-/- cells. Fig. 2A mdy2-get4-1 4 16 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 11 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Interaction observed in SGT2-/- cells. Fig. 2A get4-hsp104-2 4 16 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 11 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 10 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 12 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level association association Interaction observed in YBR137W-/- cells. Additional chaperones, only identified as HSP70s, were also seen to have bound. Fig. 2A get3-sec22-4 6 15 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 7 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined transmembrane region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 16 western blot western blot Immuno blot Immuno blot weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra unspecified role unspecified role ancillary ancillary physiological level physiological level endogenous level endogenous endogenous endogenous level yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 11 western blot western blot Immuno blot Immuno blot weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra unspecified role unspecified role ancillary ancillary physiological level physiological level endogenous level endogenous endogenous endogenous level yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast association association Fig. 1A, 1B get4-sec22-1 6 7 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined transmembrane region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 11 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Figs. 1C, 1D, Supp. Figs. 1B, 1D Interaction observed in get3-/- cells however incubation of recombinant GET3 with GET4FLAG-SEC22 bound to anti-FLAG resin resulted in SEC22 elution mdy2-sec22-1 6 7 unspecified role unspecified role ancillary ancillary in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast 16 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 12 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level in vitro In vitro association association Fig. 1C, 2C Interaction observed in get3-/- cells sgt2-sec22-1 6 7 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 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 in vitro In vitro 12 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed Q12118:p.[Arg171Ala;Arg175Ala] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain R A Wed May 02 18:00:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg171ala,arg175ala' Tue Jun 30 15:42:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg171Ala;Arg175Ala]' 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Fig. 2D, 6A, 6C, 6D Interaction observed in get3-/-, get5-/-, and get3/5 -/- cells. When GET3, GET4, and GET5 proteins were used to elute SEC22 from the column, elution was observed in the combined presence of GET3, GET4, and GET5. All other incubations containing GET3 resulted in only marginal SEC22 elution. The remaining elutions, which contained GET4 and/or GET5 without GET3, were indistinguishable from background. Increasing concentrations of GET3 stimulated SEC22 elution, increasing concentrations of GET5 progressively reduced the efficiency of SEC22 elution and even at saturating concentrations of GET3, the rate of SEC22 elution was further increased by the addition of GET4-GET5. sgt2-hsp104-1 4 8 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 12 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 Q12118:p.[Arg171Ala;Arg175Ala] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain R A Wed May 02 17:25:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg171ala,arg175ala' Tue Jun 30 19:48:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg171Ala;Arg175Ala]' 10 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level association association Interaction observed in GET3-/- cells. Additional chaperones, only identified as HSP70s, were also seen to have bound. Fig. 2E sgt2-sec22-2 6 7 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated 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 12 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain MKRDYESAKKKVEQSLNLEKTVPEQSRDADVDASQGASAGGLPDLGSLLGGGLGGLMNNPQLMQAAQKMMSNPGAMQNIQKMMQDPSIRQMAEGFASGGGTPNLSDLMNNPALRNMAGNLFGGAGAQSTDETPDNENKQ A 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Fig. 3B Interaction observed in get3-/- cells sgt2-fis1-1 6 13 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P40515:p.[Gly136_Gly137delinsLeuLeu;Gly141_Ala142delinsLeuLeu] mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain GA LL certain certain sequence position certain certain certain sequence position certain GG LL Mon Jan 22 17:14:06 GMT 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly136leu,gly137leu,gly141leu,ala142leu' Wed May 02 19:03:41 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly_gly136-137leu_leu,gly_ala141-142leu_leu' Tue Jun 30 19:38:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Gly136_Gly137delinsLeuLeu;Gly141_Ala142delinsLeuLeu]' c-terminal opsin tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range in vitro In vitro 12 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Fig. 4B Interaction observed in get5-/- cells sgt2-fis1-2 6 7 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated 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 13 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P40515:p.[Gly136_Gly137delinsLeuLeu;Gly141_Ala142delinsLeuLeu] mutation mutation certain certain sequence position certain certain certain sequence position certain GA LL certain certain sequence position certain certain certain sequence position certain GG LL Mon Jan 22 17:03:02 GMT 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly136leu,gly137leu,gly141leu,ala142leu' Wed May 02 15:13:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly_gly136-137leu_leu,gly_ala141-142leu_leu' Tue Jun 30 19:43:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Gly136_Gly137delinsLeuLeu;Gly141_Ala142delinsLeuLeu]' in vitro In vitro 12 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed Q12118:p.[Arg171Ala;Arg175Ala] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain R A Wed May 02 18:04:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg171ala,arg175ala' Tue Jun 30 17:05:51 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg171Ala;Arg175Ala]' 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast association association Fig. 4C Interaction observed in get5-/- and get3/5-/- cells. When GET3, GET4, and GEt5 proteins were used to elute SEC22 from the column, elution was observed in the combined presence of GET3, GET4, and GET5. (gly136leu,gly137leu,gly141leu,ala142leu)FIS1 was eluted with an efficiency comparable to that observed for SEC22, while FIS1 was not appreciably eluted under the same conditions sgt2-sec22-3 6 7 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated 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 12 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed Q12118:p.[Arg171Ala;Arg175Ala] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain R A Wed May 02 19:44:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg171ala,arg175ala' Tue Jun 30 18:43:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg171Ala;Arg175Ala]' 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Fig. 5A, 5B, 5C, 6A Supp. Fig. 4E Interaction observed in get3/5 -/- cells. When GET3, GET4, and GET5 proteins were used to elute SEC22 from the column, elution was observed in the combined presence of GET3, GET4, and GET5. All other incubations containing GET3 resulted in only marginal SEC22 elution. The remaining elutions, which contained GET4 and/or GET5 without GET3, were indistinguishable from background. SEC22 elution was abolished by three point mutations (GET3 L187E, F190E, and I212E) in GET3’s predicted TMD-binding site. Increasing concentrations of GET3 stimulated SEC22 elution. There was no no apparent decrease in SEC22 elution from SGT2R171A, R175A under any condition tested. get3-sgt2-1 2 15 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 12 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 16 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated in vitro In vitro 11 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated in vitro In vitro association association Fig. 6B get3-get4-5 2 15 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 16 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated in vitro In vitro 11 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated in vitro In vitro association association Fig. 6B mdy2-sec22-2 6 16 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 7 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined transmembrane region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro physical association physical association Supp. Figs. 1B, 1E Incubation of recombinant GET3 with GET4FLAG-SEC22 bound to anti-FLAG resin resulted in SEC22 elution This was abolished by three point mutations in GET3 (GET3 L187E, F190E, and I212E) sgt2-fis1-3 6 13 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined transmembrane region required to bind necessary binding region required to bind range range range range in vitro In vitro 12 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell Q12118:p.[Arg171Ala;Arg175Ala] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain R A Wed May 02 15:15:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg171ala,arg175ala' Tue Jun 30 18:58:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg171Ala;Arg175Ala]' 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Fig. 4D Interaction observed in get5-/- cells hsp104-mdy2-1 1 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell c-terminal region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 16 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell 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 Supp. Fig. 3B sgt2-hsp104-2 1 11 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell 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 Supp. Fig. 3B get4-sec22-2 6 7 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated 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 15 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 ecoli Escherichia coli (strain K12) physical association physical association Supp. Figs. 4C GET3 was first incubated with Flag-tagged SGT2 in get3/5-/- cell lysate, isolated by anti-flag tag immunoprecipitation, then by anti-His. The complex was eluted on addition of untagged GET£. sgt2-smt3-1 3 14 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell opsin tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain association association Fig. 3C get3-sec22-5 5 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast 14 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed opsin tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast 15 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 yeast Saccharomyces cerevisiae Baker's yeast association association Supp. Fig. 1A get4-sec22-3 5 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast 14 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed opsin tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast 11 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 yeast Saccharomyces cerevisiae Baker's yeast association association Supp. Fig. 1A mdy2-sec22-3 5 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 14 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed opsin tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 16 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 yeast Saccharomyces cerevisiae Baker's yeast association association Supp. Fig. 1A