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 Arf1p, Chs5p and the ChAPs are required for export of specialized cargo from the Golgi. Arf1p, Chs5p and the ChAPs are required for export of specialized cargo from the Golgi. EMBO J. (0261-4189) 2006 imex curation Trautwein M., Schindler C., Gauss R., Dengjel J., Hartmann E., Spang A. 2009-01-08 Only protein-protein interactions in vitro In vitro pull down pull down predetermined predetermined participant predetermined Trautwein M., Schindler C., Gauss R., Dengjel J., Hartmann E., Spang A. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Arf1p, Chs5p and the ChAPs are required for export of specialized cargo from the Golgi. Arf1p, Chs5p and the ChAPs are required for export of specialized cargo from the Golgi. EMBO J. (0261-4189) 2006 imex curation Trautwein M., Schindler C., Gauss R., Dengjel J., Hartmann E., Spang A. 2009-01-08 Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Trautwein M., Schindler C., Gauss R., Dengjel J., Hartmann E., Spang A. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Arf1p, Chs5p and the ChAPs are required for export of specialized cargo from the Golgi. Arf1p, Chs5p and the ChAPs are required for export of specialized cargo from the Golgi. EMBO J. (0261-4189) 2006 imex curation Trautwein M., Schindler C., Gauss R., Dengjel J., Hartmann E., Spang A. 2009-01-08 Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Trautwein M., Schindler C., Gauss R., Dengjel J., Hartmann E., Spang A. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Arf1p, Chs5p and the ChAPs are required for export of specialized cargo from the Golgi. Arf1p, Chs5p and the ChAPs are required for export of specialized cargo from the Golgi. EMBO J. (0261-4189) 2006 imex curation Trautwein M., Schindler C., Gauss R., Dengjel J., Hartmann E., Spang A. 2009-01-08 Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast crosslink cross-linking study crosslink predetermined predetermined participant predetermined Trautwein M., Schindler C., Gauss R., Dengjel J., Hartmann E., Spang A. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation chs6_yeast Chitin biosynthesis protein CHS6 CHS6 CSD3 YJL099W J0838 Protein CSD3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MNLFWPSETKKQNEIPGGDYTPGNSPSVQKGYQFLNRDIFKSCPRIMERQFGECLHNRTHLIKDLISSGNVGLGPIEIVHMSYLNKHEKEEFGEYFYVTGIEVSGPAMPVEFLEVLKSSKRISKNISNNIILTYCCFNFFSNLDIRIRYDADDTFQTTAIDCNKETTDLTMTEKMWEETFASSVIRAIITNTNPELKPPGLVECPFYVGKDTISSCKKIIELLCRFLPRSLNCGWDSTKSMQATIVNNYLMYSLKSFIAITPSLVDFTIDYLKGLTKKDPIHDIYYKTAMITILDHIETKELDMITILNETLDPLLSLLNDLPPRDADSARLMNCMSDLLNIQTNFLLNRGDYELALGVSNTSTELALDSFESWYNLARCHIKKEEYEKALFAINSMPRLRKNDGHLETMYSRFLTSNYYKKPLNGTREHYDLTAMEFTNLSGTLRNWKEDELKRQIFGRIAMINEKKIGYTKEIWDDIAIKLGPICGPQSVNLINYVSPQEVKNIKNINLIARNTIGKQLGWFSGKIYGLLMEIVNKIGWNGLLNIRTEAFMMETEFYQASNNIIDENGHIPMESRKKRFCEGWLDDLFLDLYQDLKLSKISLSNKDEKHSGLEWELLGLIMLRTWHWEDAVACLRTSIVARFDPVSCQQLLKIYLQPPKNIQEVTLLDTDTIISLLIKKISYDCRYYNYCQIFNLQLLEKLCNELGTHILRNKILLQPSIGDEIMVMIDAMLAWIADLDHTVQP BD92DADA3D5396BC BD92DADA3D5396BC bch1_yeast Protein BCH1 YMR237W YM9959.19 BCH1 BUD7 and CHS6 homolog 1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MLSQTSIPEVKEDVIGYALHQRRARVGQFQDLGPPDLITLIKSLPSSSSTTTATASANDNGATSNINGQDPTTIVTELHSHDKLKGQIGTFFYCMGIDTSDPTSITIFAKKITDLFLDTPQIWFGKKKHFHVSKISISSWNAFRKYDVNIIVHIPGTVQTYIINSDGEQSQLPSVAEASSGRNSQDLNVNMIWAETFMSGIVRDIMIMKDNRADGESQNLVETLIFNPFTSGELEDVANNFIKLFPLVYEKGVYLDAPTHVLNPSLTNNYLVETLVEIVRLTKSLEACRKMLKKLIEIHPEAVIILIRVYFACDLEIDAVDLINEQLNSPSSFLADDSKTSHIQLIFKSELLSIQSEFLLDVKRDYKLAKEVAMEAVNCAPNEFKTWYLLTRIYIKLNDMSNALLSLNACPMSQVKEKYVLRRIAPITSDENLHLPLPLDASIEEISSLNPMDVQLEQKSADPNLVNLSASSLKSTFQLAYKLLTEIVQITGWEQLLKYRSKIFVMEDEYQGSTSSIDEAEVRGNDISKMRSKRLCERWLDNLFMLLYEDLKTYTDWQSEQLYFDAQNSKYHKLTVEWELFGLCAKRLGHLPEAAKAFQIGLSQRFSPVCAKNLLQFYIDEHKRIRRDSVSANSELTSSQILSSINDIDSSIIDLVVKICCWNHRWYIEFSIILIDALSVAVQDMGITKVHNEIASRFSDPVAQLIDDNILNFLKNFTNDTFDN 3D9B80FDE9CF202B 3D9B80FDE9CF202B bch2_yeast Protein BCH2 YKR027W BUD7 and CHS6 homolog 2 BCH2 FMP50 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSFLWGSTKSKKGKNKKAAGSLPSGVVPQQRVKPTRKNVPIDYPRTLEKVHGESLIFRTSLLSELVSTGKSGIGPPDLIHCTELDKFHDEKIGEFFYITGIDASSVSMPIAFLKLIKWNDGKKLKSASLKNDDITTYCTFNIFQKLDIRLRYESEDVYQVNIVDCLNGNNEIPLSDLIWEETFVSCCIRSVIINSDFERKIPGLVELPFVFENRCASDYKRVIDSLCKFLPRFLECGWDSTKSVYATILNNYLTESLLVFLSITPEFITDYAIQVLDNLMTNDPSNSRYYAIVIISIMERSNDRDVEMIKRIHEILDLLLPVLYGLPSDEPYISDLINCITDVLSIQARFLLNNNDYELSLSISTLATNLSSDNFESWYLLSKGYIFSQQYDKALLSINSMPCLAEYDIVKQAQINAFKFYMNYYKAPLCHSREHCTMTSHELNHLMNIMHYENELELKTIIFGRTVMPNESKYGCIEEIWNKSCLELGPICGPQSDNLINFVSQQEVNTVGDMLLLKRSKETRQESWFIKQVRLLLMELVARIGWNALLQLRSDVFVMESKFKMIESSDKLSTELRQKRLCQRWFDAMFLDVYEDLSISTSSQENKATAKYSGLEWELLGLTLLRVSDLPDAVACLRTSILARFDPISCHHLLNFYLTMDFNDEFMRRFDVDIILDLLVKLISFRIRFYDRFQIFSLQVLRKLEGQLGSEIIKNKIINSPYGQAGITSVIDYMLECLSKNRNEACLAYERPLPDLPSTIKPLAD 53A4A700682287E8 chs3_yeast Chitin synthase 3 CHS3 CAL1 DIT101 KIT2 YBR023C CSD2 YBR0305 Chitin-UDP acetyl-glucosaminyl transferase 3 Class-IV chitin synthase 3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MTGLNGDDPDDYYLNLNQDEESLLRSRHSVGSGAPHRQGSLVRPERSRLNNPDNPHFYYAQKTQEQMNHLDVLPSSTGVNPNATRRSGSLRSKGSVRSKFSGRETDSYLLQDMNTTDKKASVKISDEGVAEDEFDKDGDVDNFEESSTQPINKSIKPLRKETNDTLSFWQMYCYFITFWAPAPILAFCGMPKKERQMAWREKVALISVILYIGAIVAFLTFGFTKTVCSSSKLRLKNNEVSTEFVVINGKAYELDTSSRSGIQDVEVDSDTLYGPWSDAGKDASFLFQNVNGNCHNLITPKSNSSIPHDDDNNLAWYFPCKLKNQDGSSKPNFTVENYAGWNCHTSKEDRDAFYGLKSKADVYFTWDGIKNSSRNLIVYNGDVLDLDLLDWLEKDDVDYPVVFDDLKTSNLQGYDLSLVLSNGHERKIARCLSEIIKVGEVDSKTVGCIASDVVLYVSLVFILSVVIIKFIIACYFRWTVARKQGAYIVDNKTMDKHTNDIEDWSNNIQTKAPLKEVDPHLRPKKYSKKSLGHKRASTFDLLKKHSSKMFQFNESVIDLDTSMSSSLQSSGSYRGMTTMTTQNAWKLSNENKAVHSRNPSTLLPTSSMFWNKATSSPVPGSSLIQSLDSTIIHPDIVQQPPLDFMPYGFPLIHTICFVTCYSEDEEGLRTTLDSLSTTDYPNSHKLLMVVCDGLIKGSGNDKTTPEIALGMMDDFVTPPDEVKPYSYVAVASGSKRHNMAKIYAGFYKYDDSTIPPENQQRVPIITIVKCGTPAEQGAAKPGNRGKRDSQIILMSFLEKITFDERMTQLEFQLLKNIWQITGLMADFYETVLMVDADTKVFPDALTHMVAEMVKDPLIMGLCGETKIANKAQSWVTAIQVFEYYISHHQAKAFESVFGSVTCLPGCFSMYRIKSPKGSDGYWVPVLANPDIVERYSDNVTNTLHKKNLLLLGEDRFLSSLMLKTFPKRKQVFVPKAACKTIAPDKFKVLLSQRRRWINSTVHNLFELVLIRDLCGTFCFSMQFVIGIELIGTMVLPLAICFTIYVIIFAIVSKPTPVITLVLLAIILGLPGLIVVITATRWSYLWWMCVYICALPIWNFVLPSYAYWKFDDFSWGDTRTIAGGNKKAQDENEGEFDHSKIKMRTWREFEREDILNRKEESDSFVA EBF9227DC30D3EA4 chs5_yeast Chitin biosynthesis protein CHS5 CHS5 CAL3 YLR330W L8543.18 Protein CAL3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSSVDVLLTVGKLDASLALLTTQDHHVIEFPTVLLPENVKAGSIIKMQVSQNLEEEKKQRNHFKSIQAKILEKYGTHKPESPVLKIVNVTQTSCVLAWDPLKLGSAKLKSLILYRKGIRSMVIPNPFKVTTTKISGLSVDTPYEFQLKLITTSGTLWSEKVILRTHKMTDMSGITVCLGPLDPLKEISDLQISQCLSHIGARPLQRHVAIDTTHFVCNDLDNEESNEELIRAKHNNIPIVRPEWVRACEVEKRIVGVRGFYLDADQSILKNYTFPPVNEEELSYSKENEPVAEVADENKMPEDTTDVEQVASPNDNESNPSEAKEQGEKSGHETAPVSPVEDPLHASTALENETTIETVNPSVRSLKSEPVGTPNIEENKADSSAEAVVEEPNEAVAESSPNEEATGQKSEDTDTHSNEQADNGFVQTEEVAENNITTESAGENNEPADDAAMEFGRPEAEIETPEVNESIEDANEPAEDSNEPVEDSNKPVKDSNKPVEDSNKPVEDSNKPVEDSNKPVEDANEPVEDTSEPVEDAGEPVQETNEFTTDIASPRHQEEDIELEAEPKDATESVAVEPSNEDVKPEEKGSEAEDDINNVSKEAASGESTTHQKTEASASLESSAVTEEQETTEAEVNTDDVLSTKEAKKNTGNSNSNKKKNKKNKKKGKKK FA92741B862814C2 FA92741B862814C2 glo3_yeast ADP-ribosylation factor GTPase-activating protein GLO3 YER122C GLO3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSNDEGETFATEQTTQQVFQKLGSNMENRVCFDCGNKNPTWTSVPFGVMLCIQCSAVHRNMGVHITFVKSSTLDKWTINNLRRFKLGGNHKARDFFLKNNGKQLLNTANVDAKTKYTSPVAKKYKIHLDKKVQKDMELYPSELVLNGQDSSDSPLDTDSDASRSTSKENSVDDFFSNWQKPSSNSSSKLNVNTGSLAPKNNTTGSTPKTTVTKTRSSILTASRKKPVLNSQDKKKHSILSSSRKPTRLTAKKVDKSQAEDLFDQFKKEAQQEKEDEFTNSSSSTKIRQNDYDSQFMNNSKGNNNNSIDDINTQPDEFNDFLNDTSNSFDTTRKEQQDTLTPKFAKLGFGMTMNDANDLAKQQKESQKIAQGPRYTGRIAERYGTQKAISSDQLFGRGSFDEAANREAHDKLKTFDNATSISSSSYFGEDKEVDEFGNPINSSGSGAGNFDGRNSNNGFIDFNASADDELQMLRDVVEQGAEKLGSYLRDYLRK CDB588BF1351A042 bud7_yeast Bud site selection protein 7 BUD7 YOR299W protein protein yeast Saccharomyces cerevisiae Baker's yeast MITQNSIPEVKEDFIGYALHERRIRLPQFQDLGPADLVTLTKYLPTSSNTNAINSTSRNGAAIIQSPAAVVADDSAASMATNGDASDTAVTTNYTNASIYSSSRNANDGAPMVAELHPLDKLKDEVGTFFYSMGVDTSGPTSIAIFLKEISEVISEKPQVWFGRKKTFNVARISFSTWNAFRRCDINVVVHIPGSIQNFIVDCNGESQNIEMCADYDLIWAETFVSGVVRSIMLMKENAEEGELQNLVETLILNPFTAGQIDDVPEMFIDLFPIVYHKGPLLGAPYYITNVTNTNNYLVETLVEIVKLTRNVSRAEIMLKNLATDNPEAIIILIKIFLVCDQELDAIKLTYDMLSQDKIINNTNNRMDYKSELLCLQAQFLIDKRQDYSLAQNIAQEAVNCSPSEFRPWYLLSKVYVKLNDIENALLILNSCPMSPLKEKYVLKRVAPLPSNNSLHLPLPIDVVLDEVTSLNPQDVQNEHRSADPMLVNLAASNLKSTFQLAYRLLTEIVQITGWENLLKYRSNIFVMEEEYQKSSSSLPKDVNKQEEQPLRAKRLCERWLDNLFMLLYEDLKMYTLWQTEQLYMDAQNNNHNKLTFEWELFGLCARRLGHFPEAAKAFQNGLSQRFSSRCARKLLEYCINERQRVKNFINSPNSHDMVPEIVSSRIRELDNSIIDLCVKICCWNHRWYTEFSISLLDCLSVVIQDMSLTKVSNEISSRYPETVLNLVQENLLNFFTTCTIGCYDA 93E6DF36F5A5C1F2 93E6DF36F5A5C1F2 arf1_yeast ADP-ribosylation factor 1 ARF1 D1244 YDL192W protein protein yeast Saccharomyces cerevisiae Baker's yeast MGLFASKLFSNLFGNKEMRILMVGLDGAGKTTVLYKLKLGEVITTIPTIGFNVETVQYKNISFTVWDVGGQDRIRSLWRHYYRNTEGVIFVVDSNDRSRIGEAREVMQRMLNEDELRNAAWLVFANKQDLPEAMSAAEITEKLGLHSIRNRPWFIQATCATSGEGLYEGLEWLSNSLKNST 796859026AC63CC0 796859026AC63CC0 chs3-bud7 4 8 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 11 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F9B EMBO-Journal mint chs3-bch1 4 8 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F9B EMBO-Journal We could also detect an interaction of Chs3p with Bch1p-9myc and Bch2p-9myc (Figure 9B), indicating that they might also be involved in export of Chs3p from the TGN. mint chs3-bch2 4 8 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F9B EMBO-Journal We could also detect an interaction of Chs3p with Bch1p-9myc and Bch2p-9myc (Figure 9B), indicating that they might also be involved in export of Chs3p from the TGN. mint chs3-chs6 4 8 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F9A EMBO-Journal Chs6p-9myc was crosslinked to Chs3p (Figure 9A). mint arf1-glo3-3 1 10 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 12 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified P11076:p.Gln71Leu mutation increasing interaction mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Q L Tue Jul 12 10:53:16 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation increasing' Wed May 02 14:55:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln71leu' Tue Jun 30 17:23:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln71Leu' range MINT-1788005: original sequence: 71-71 : ISFTVWDVGGQDRIRSLWRH Stoichiometry: 1.0 physical association physical association F2A EMBO-Journal As expected from their biological functions, the coatomer complex and the ARF-GAP Glo3p bound preferentially to Arf1p-GTP, whereas the ARF-GEF Gea2p bound only to Arf1p-GDP (Figure 2A). mint arf1-chs5-2 1 9 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 12 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified P11076:p.Gln71Leu mutation increasing interaction mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Q L Tue Jul 12 10:10:43 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation increasing' Wed May 02 19:03:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln71leu' Tue Jun 30 16:38:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln71Leu' range MINT-1788020: original sequence: 71-71 : ISFTVWDVGGQDRIRSLWRH Stoichiometry: 1.0 physical association physical association F2B EMBO-Journal We found that Chs5p bound specifically to the activated form of Arf1p (Figure 2B) mint arf1-chs6 2 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 12 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 physical association physical association F2C F7A EMBO-Journal To test whether the other ChAPs can also interact with Arf1p, we performed co-immunoprecipitations with alpha-Arf1p antibodies on lysates from strains where the ChAPs were appended chromosomally with a 9myc-tag. The proteins were expressed from their endogenous promotor and were fully functional. A signal for Bch1p-9myc, Bud7p-9myc, Chs6p-9myc and Bch2p-9myc was obtained in the precipitate (Figures 2C and 7A), indicating that all ChAPs can associate with Arf1p. mint arf1-bch2 2 12 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F2C F7A EMBO-Journal To test whether the other ChAPs can also interact with Arf1p, we performed co-immunoprecipitations with alpha-Arf1p antibodies on lysates from strains where the ChAPs were appended chromosomally with a 9myc-tag. The proteins were expressed from their endogenous promotor and were fully functional. A signal for Bch1p-9myc, Bud7p-9myc, Chs6p-9myc and Bch2p-9myc was obtained in the precipitate (Figures 2C and 7A), indicating that all ChAPs can associate with Arf1p. mint arf1-bud7 2 12 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 11 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F2C F7A EMBO-Journal To test whether the other ChAPs can also interact with Arf1p, we performed co-immunoprecipitations with alpha-Arf1p antibodies on lysates from strains where the ChAPs were appended chromosomally with a 9myc-tag. The proteins were expressed from their endogenous promotor and were fully functional. A signal for Bch1p-9myc, Bud7p-9myc, Chs6p-9myc and Bch2p-9myc was obtained in the precipitate (Figures 2C and 7A), indicating that all ChAPs can associate with Arf1p. mint arf1-bch1 2 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 12 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 physical association physical association F2C F7A EMBO-Journal To test whether the other ChAPs can also interact with Arf1p, we performed co-immunoprecipitations with alpha-Arf1p antibodies on lysates from strains where the ChAPs were appended chromosomally with a 9myc-tag. The proteins were expressed from their endogenous promotor and were fully functional. A signal for Bch1p-9myc, Bud7p-9myc, Chs6p-9myc and Bch2p-9myc was obtained in the precipitate (Figures 2C and 7A), indicating that all ChAPs can associate with Arf1p. mint chs5-chs6-8 2 9 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F7B EMBO-Journal To test if the other ChAPs would also bind to Chs5p, we performed precipitations with Chs5p antiserum on the same lysates that were used to detect the Arf1p interaction (Figure 7). All ChAPs co-immunoprecipitated with Chs5p (Figure 7B, lane 5). mint chs5-bch1-20 2 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 9 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 physical association physical association F7B EMBO-Journal To test if the other ChAPs would also bind to Chs5p, we performed precipitations with Chs5p antiserum on the same lysates that were used to detect the Arf1p interaction (Figure 7). All ChAPs co-immunoprecipitated with Chs5p (Figure 7B, lane 5). mint chs5-bch2-7 2 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 9 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 physical association physical association F7B EMBO-Journal To test if the other ChAPs would also bind to Chs5p, we performed precipitations with Chs5p antiserum on the same lysates that were used to detect the Arf1p interaction (Figure 7). All ChAPs co-immunoprecipitated with Chs5p (Figure 7B, lane 5). mint chs5-bud7-3 2 9 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 11 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F7B EMBO-Journal To test if the other ChAPs would also bind to Chs5p, we performed precipitations with Chs5p antiserum on the same lysates that were used to detect the Arf1p interaction (Figure 7). All ChAPs co-immunoprecipitated with Chs5p (Figure 7B, lane 5). mint bch1-bch2 3 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level ha tagged ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4 EMBO-Journal When polyclonal AU5 antibodies were used for precipitation, all the three other ChAPs were detected (Figure 4). Co-precipitation was specific, because no ChAPs were detected from control strains lacking the AU5 tag. mint bch1-bud7 3 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 11 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4 EMBO-Journal When polyclonal AU5 antibodies were used for precipitation, all the three other ChAPs were detected (Figure 4). Co-precipitation was specific, because no ChAPs were detected from control strains lacking the AU5 tag. mint bch1-chs6 3 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level gfp tagged gfp tagged 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 Stoichiometry: 1.0 physical association physical association F4 EMBO-Journal When polyclonal AU5 antibodies were used for precipitation, all the three other ChAPs were detected (Figure 4). Co-precipitation was specific, because no ChAPs were detected from control strains lacking the AU5 tag. mint bud7-bch1 3 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 11 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4 EMBO-Journal A similar result was obtained for Bud7p-9myc that was precipitated with monoclonal myc antibodies. Again, Bch2p, Chs6p and Bch1p were detected when Bud7p-9myc was precipitated and were absent when control lysates lacking the myc epitope were used. mint bud7-bch2 3 11 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level ha tagged ha tagged ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4 EMBO-Journal A similar result was obtained for Bud7p-9myc that was precipitated with monoclonal myc antibodies. Again, Bch2p, Chs6p and Bch1p were detected when Bud7p-9myc was precipitated and were absent when control lysates lacking the myc epitope were used. mint bud7-chs6 3 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level gfp tagged gfp tagged 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 Stoichiometry: 1.0 11 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4 EMBO-Journal A similar result was obtained for Bud7p-9myc that was precipitated with monoclonal myc antibodies. Again, Bch2p, Chs6p and Bch1p were detected when Bud7p-9myc was precipitated and were absent when control lysates lacking the myc epitope were used. mint