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/ Structural and functional studies of Bud23-Trm112 reveal 18S rRNA N7-G1575 methylation occurs on late 40S precursor ribosomes. Structural and functional studies of Bud23-Trm112 reveal 18S rRNA N7-G1575 methylation occurs on late 40S precursor ribosomes. Proceedings of the National Academy of Sciences of the United States of America 2014 imex curation Létoquart J., Huvelle E., Wacheul L., Bourgeois G., Zorbas C., Graille M., Heurgué-Hamard V., Lafontaine DL. denis.lafontaine@ulb.ac.be, marc.graille@polytechnique.edu, valerie.heurgue@ibpc.fr. 18-Apr-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation ecoli Escherichia coli (strain K12) pull down pull down western blot western blot Immuno blot Immuno blot Létoquart J., Huvelle E., Wacheul L., Bourgeois G., Zorbas C., Graille M., Heurgué-Hamard V., Lafontaine DL. Abs were raised against the complex, then used to identify the two proteins when separated on SDS-PAGE. denis.lafontaine@ulb.ac.be, marc.graille@polytechnique.edu, valerie.heurgue@ibpc.fr. Accepted 2017-MAR-13 AT 11:34 GMT AT 11:34 GMT by MGT Proceedings of the National Academy of Sciences of the United States of America 2014 Only protein-protein interactions imex curation imex curation No, its Supp Fig 2B - the figure is there to show expression levels, but the complex was first purified by pulldown before they ran the gel so it also works as a proof of interaction. M=As they appear to be arguing that the mutants affect expression levels I've not included them ,though in fact they could equally well be affecting the interaction. Structural and functional studies of Bud23-Trm112 reveal 18S rRNA N7-G1575 methylation occurs on late 40S precursor ribosomes. Structural and functional studies of Bud23-Trm112 reveal 18S rRNA N7-G1575 methylation occurs on late 40S precursor ribosomes. Proceedings of the National Academy of Sciences of the United States of America 2014 imex curation Létoquart J., Huvelle E., Wacheul L., Bourgeois G., Zorbas C., Graille M., Heurgué-Hamard V., Lafontaine DL. denis.lafontaine@ulb.ac.be, marc.graille@polytechnique.edu, valerie.heurgue@ibpc.fr. 18-Apr-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Létoquart J., Huvelle E., Wacheul L., Bourgeois G., Zorbas C., Graille M., Heurgué-Hamard V., Lafontaine DL. denis.lafontaine@ulb.ac.be, marc.graille@polytechnique.edu, valerie.heurgue@ibpc.fr. Accepted 2017-MAR-13 AT 11:34 GMT AT 11:34 GMT by MGT Proceedings of the National Academy of Sciences of the United States of America 2014 Only protein-protein interactions imex curation imex curation No, its Supp Fig 2B - the figure is there to show expression levels, but the complex was first purified by pulldown before they ran the gel so it also works as a proof of interaction. M=As they appear to be arguing that the mutants affect expression levels I've not included them ,though in fact they could equally well be affecting the interaction. Structural and functional studies of Bud23-Trm112 reveal 18S rRNA N7-G1575 methylation occurs on late 40S precursor ribosomes. Structural and functional studies of Bud23-Trm112 reveal 18S rRNA N7-G1575 methylation occurs on late 40S precursor ribosomes. Proceedings of the National Academy of Sciences of the United States of America 2014 imex curation Létoquart J., Huvelle E., Wacheul L., Bourgeois G., Zorbas C., Graille M., Heurgué-Hamard V., Lafontaine DL. denis.lafontaine@ulb.ac.be, marc.graille@polytechnique.edu, valerie.heurgue@ibpc.fr. 18-Apr-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Létoquart J., Huvelle E., Wacheul L., Bourgeois G., Zorbas C., Graille M., Heurgué-Hamard V., Lafontaine DL. denis.lafontaine@ulb.ac.be, marc.graille@polytechnique.edu, valerie.heurgue@ibpc.fr. Accepted 2017-MAR-13 AT 11:34 GMT AT 11:34 GMT by MGT Proceedings of the National Academy of Sciences of the United States of America 2014 Only protein-protein interactions imex curation imex curation No, its Supp Fig 2B - the figure is there to show expression levels, but the complex was first purified by pulldown before they ran the gel so it also works as a proof of interaction. M=As they appear to be arguing that the mutants affect expression levels I've not included them ,though in fact they could equally well be affecting the interaction. bud23_yeast 18S rRNA (guanine(1575)-N(7))-methyltransferase Bud site selection protein 23 YCR047C YCR47C BUD23 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSRPEELAPPEIFYNDSEAHKYTGSTRVQHIQAKMTLRALELLNLQPCSFILDIGCGSGLSGEILTQEGDHVWCGLDISPSMLATGLSRELEGDLMLQDMGTGIPFRAGSFDAAISISAIQWLCNADTSYNDPKQRLMRFFNTLYAALKKGGKFVAQFYPKNDDQVDDILQSAKVAGFSGGLVVDDPESKKNKKYYLVLSSGAPPQGEEQVNLDGVTMDEENVNLKKQLRQRLKGGKDKESAKSFILRKKELMKRRGRKVAKDSKFTGRKRRHRF A8011A097C461900 A8011A097C461900 s-adenosyl-l-methionine S adenosyl Methionine small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) dhr1_yeast Probable ATP-dependent RNA helicase DHR1 ECM16 DHR1 YMR128W YM9553.04 DEAH box RNA helicase DHR1 Extracellular mutant protein 16 protein protein yeast Saccharomyces cerevisiae Baker's yeast MGTYRKRFNEKARSGHMAKLKELKRIRNKQFTRQDENDERVENPDSAPAESSTTEPNANAEILEPLTEEEKKMKKRKLQELFTPKESKVSRLKKKRLDKFIEHQLKREERKTIIGKLQDYKIDTSLLTSSKRLGEGRQTKKEEFKEALSLERQGRGNEQTNEILYEEYEPKVWDEYGEGGSSEDDDGEDDFEASFGSMPKPTDNEEKKSSGFIDHRPAKFGGSGLSFGFSNIKVINKESKTPKKKYNWRQRVEMEELKKHGKEDEMDFDTTSEDDDEEEDQEEEDKMHPSENPLEEVESADSETGSEKFDQNDVANEFKDWANQEIKKLEGRDQELVTPTLNIDYKPIIRKEDLDDGLQEAYVPINENSTRKAFYVEVSRSDEIQKARIQLPVFGEEHKIMEAIHHNDVVIICGETGSGKTTQVPQFLYEAGFGAEDSPDYPGMVGITQPRRVAAVSMAERVANELGDHGHKVGYQIRFDSTAKEDTKVKFMTDGVLLREMMHDFKLTKYSSIIIDEAHERNINTDILIGMLSRCVRLRAKLHKENPIEHKKLKLIIMSATLRVSDFSENKTLFPIAPPVLQVDARQFPVSIHFNRRTAFNYTDEAFRKTCKIHQKLPPGAILVFLTGQQEITHMVKRLRKEFPFKKNSKYNKDLETPVSKMGINSKTTDLEAEDIDFSVQVIDQDKFKSAIRYEEDEGNSGNGEDEEDEEEEGFEEVLTEGQTANDPLYVLPLYSLLPTKEQMRVFQKPPQGSRLCIVATNVAETSLTIPGVRYVVDSGRSKERKYNESNGVQSFEVGWVSKASANQRSGRAGRTGPGHCYRLYSSAVFEHDFEQFSKPEILRMPVESIVLQMKSMAIHNIINFPFPTPPDRVALSKAIQLLQYLGALDNKEMITEDGKKMSLFPLSPRFSKMLLVSDEKACLPYIVAIVSALSVGDPFINEFELGINEISRKPNPDENLDDKIREHDESTPGMDPELKKELRSKFYKSRSQFSKLDKFSDVFRLLSVVSAMDYVPKEQKEIFMKKNFLRGKLMEEIVKLRKQLMYIIKSNTSKENIAVVIRNEDLKSDIPSVIQIKLLKQMICAGFVDHVAVRADVLFPDDAKITNRTSIINIPYIPVLATRTPNIEDCFVYIHPTSILNNLGEMPPKYMLYYSLHLGGNNKTRMNTLCDIASTPLANIARKGLLLTYSKPLTGQGLKTVNLSPTERYCYVVPRFGSTVDNDLKIGWDLNPIAVHQKKQKGQWTVIKFITRKGFQTITGEEKEKK 576DAAE6D934CC77 tr112_yeast Multifunctional methyltransferase subunit TRM112 YNR046W N3445 TRM112 tRNA methyltransferase 112 eRF1 methyltransferase subunit TRM112 protein protein yeast Saccharomyces cerevisiae Baker's yeast MKFLTTNFLKCSVKACDTSNDNFPLQYDGSKCQLVQDESIEFNPEFLLNIVDRVDWPAVLTVAAELGNNALPPTKPSFPSSIQELTDDDMAILNDLHTLLLQTSIAEGEMKCRNCGHIYYIKNGIPNLLLPPHLV 3CFB2ABDDFB0E38A bud23-trm112-1 3 7 unspecified role unspecified role neutral component neutral component bud23 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role neutral component neutral component 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 trm112 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 13 10 direct interaction direct interaction Supp Table 6 2A Rworking 0.191 Rfree 0.242 bud23-trm112-2 1 7 unspecified role unspecified role prey prey bud23 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined trm112 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction Supp Fig. 2B bud23-trm112-3 3 5 cofactor cofactor coenzyme neutral component neutral component 7 unspecified role unspecified role neutral component neutral component bud23 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role neutral component neutral component 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 trm112 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Supp Table 6 2.12A Rworking 0.178 Rfree 0.229 bud23-trm112-4 2 7 unspecified role unspecified role neutral component neutral component bud23 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role neutral component neutral component 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 trm112 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 3A bud23-ecm16-1 2 6 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component bud23 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role neutral component neutral component trm112 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 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 Fig. 3A