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/ The Caenorhabditis elegans maternal-effect sterile proteins, MES-2, MES-3, and MES-6, are associated in a complex in embryos. The Caenorhabditis elegans maternal-effect sterile proteins, MES-2, MES-3, and MES-6, are associated in a complex in embryos. Proceedings of the National Academy of Sciences of the United States of America 2001 imex curation Xu L., Fong Y., Strome S., Xu L., Fong Y., Strome S. sstrome@bio.indiana.edu 05-Sep-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Xu L., Fong Y., Strome S., Xu L., Fong Y., Strome S. sstrome@bio.indiana.edu Accepted 2017-JUL-31 AT 14:18 BST AT 14:18 BST by MGT Proceedings of the National Academy of Sciences of the United States of America 2001 imex curation The Caenorhabditis elegans maternal-effect sterile proteins, MES-2, MES-3, and MES-6, are associated in a complex in embryos. The Caenorhabditis elegans maternal-effect sterile proteins, MES-2, MES-3, and MES-6, are associated in a complex in embryos. Proceedings of the National Academy of Sciences of the United States of America 2001 imex curation Xu L., Fong Y., Strome S., Xu L., Fong Y., Strome S. sstrome@bio.indiana.edu 05-Sep-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation caeel-embryo Caenorhabditis elegans embryo embryo Embryo Defined as the development stage prior to the initiation of tissue differentiation. molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Xu L., Fong Y., Strome S., Xu L., Fong Y., Strome S. no I've not added the density gradient as from whole cell extract sstrome@bio.indiana.edu Accepted 2017-JUL-31 AT 14:18 BST AT 14:18 BST by MGT Proceedings of the National Academy of Sciences of the United States of America 2001 imex curation The Caenorhabditis elegans maternal-effect sterile proteins, MES-2, MES-3, and MES-6, are associated in a complex in embryos. The Caenorhabditis elegans maternal-effect sterile proteins, MES-2, MES-3, and MES-6, are associated in a complex in embryos. Proceedings of the National Academy of Sciences of the United States of America 2001 imex curation Xu L., Fong Y., Strome S., Xu L., Fong Y., Strome S. sstrome@bio.indiana.edu 05-Sep-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation caeel-embryo Caenorhabditis elegans embryo embryo Embryo Defined as the development stage prior to the initiation of tissue differentiation. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Xu L., Fong Y., Strome S., Xu L., Fong Y., Strome S. sstrome@bio.indiana.edu Accepted 2017-JUL-31 AT 14:18 BST AT 14:18 BST by MGT Proceedings of the National Academy of Sciences of the United States of America 2001 imex curation mes2_caeel Histone-lysine N-methyltransferase mes-2 E(z) homolog Maternal-effect sterile protein 2 R06A4.7 mes-2 protein protein caeel Caenorhabditis elegans MSNSEPSTSTPSGKTKKRGKKCETSMGKSKKSKNLPRFVKIQPIFSSEKIKETVCEQGIEECKRMLKGHFNAIKDDYDIRVKDELDTDIKDWLKDASSSVNEYRRRLQENLGEGRTIAKFSFKNCEKYEENDYKVSDSTVTWIKPDRTEEGDLMKKFRAPCSRIEVGDISPPMIYWVPIEQSVATPDQLRLTHMPYFGDGIDDGNIYEHLIDMFPDGIHGFSDNWSYVNDWILYKLCRAALKDYQGSPDVFYYTLYRLWPNKSSQREFSSAFPVLCENFAEKGFDPSSLEPWKKTKIAEGAQNLRNPTCYACLAYTCAIHGFKAEIPIEFPNGEFYNAMLPLPNNPENDGKMCSGNCWKSVTMKEVSEVLVPDSEEILQKEVKIYFMKSRIAKMPIEDGALIVNIYVFNTYIPFCEFVKKYVDEDDEESKIRSCRDAYHLMMSMAENVSARRLKMGQPSNRLSIKDRVNNFRRNQLSQEKAKRKLRHDSLRIQALRDGLDAEKLIREDDMRDSQRNSEKVRMTAVTPITACRHAGPCNATAENCACRENGVCSYMCKCDINCSQRFPGCNCAAGQCYTKACQCYRANWECNPMTCNMCKCDAIDSNIIKCRNFGMTRMIQKRTYCGPSKIAGNGLFLLEPAEKDEFITEYTGERISDDEAERRGAIYDRYQCSYIFNIETGGAIDSYKIGNLARFANHDSKNPTCYARTMVVAGEHRIGFYAKRRLEISEELTFDYSYSGEHQIAFRMVQTKERSEKPSRPKSQKLSKPMTSE 91ABEBAD94A1D51E mes3_caeel Polycomb protein mes-3 mes-3 F54C1.3 Maternal-effect sterile protein 3 protein protein caeel Caenorhabditis elegans MTPATAEVKVGGRKRKNDEPVFIKTKQSRSARREEEKENLLNKSLPSTPTSSEAGSSRESSNPVTSSSRRKNPPTKLENIQKTLPTCSDGLEIRNYVKKYGLPEDNKFLVRNVFDKQLLFGKKYVCRRRVIKSIDEFFPRLKTDAHRENGLYNFCTLQFMKISSWGSSMDEKIYVTSAAIVQSYVIIVDDNDSVTCIENGISVIPMANRTLPSISYNSSNVKNITLTGPELKKAKNVYLAVIVKRKTPKLGENRVGTRGKSTRASFETAASSENFQQLVRFGCTLMYDKDAEINCLNDGVQRIILLDREEKPAYLKIFGKADEPELENAWMNNVKHLEFISDTNYNKEDSHLARLIFSSNTHNYFDRPDISKFSPWQPRSRRTTLKSGISVEPGTSCDEDCGSQYEDTITLKQLLLGTKKLCGRRKRHLTRLTSRWPCFSVDPTKINSYFNETGVTLLENSCSYRPGTQRSIPIKPTRVIRVREETPVSVPIEVITIEDDSDDSPCFSARNRAPGGTRKYGLDFIQSSGHMPYIRYVYMNRANDLVPQLNGEENGKIEGLMEAYPMNNAHMYSDIEFLRKEKLNLPKNAQKYGEMTILDYPLTKTETDEYNKKMSVMNVKDFAVKPRKPIPTVGNTTLSLDLEGCPISTFSEIFFPSGKTELFNILCRHFYSEAGQRPGCFSAQWKSRVLTGFINKYHQYIFDMKLNSLFEKQINLIAMVSPDWKLEDFELPMVRYKQLKKEWKNRQRDIPSSN B7B3EFCFAC0FA512 mes6_caeel Polycomb protein mes-6 mes-6 C09G4.5 Maternal-effect sterile protein 6 ESC homolog protein protein caeel Caenorhabditis elegans MEHTKKFKSLNLHGFDRSTEDYGKRPFVLTAKLLEDQKKAIYGCAFNQYAGIDEEQAVATVGGSFLHMYSVPIDINNIELQWSCNFPTDKSSKVEREESLFTVTWCYDTYEAENDRNPFKVVTGGTLGHIYVIDYVSRKLSNRLRSVGWEINDIRTCPANSNLIVCASSDQSIRIHHIRNEACLIVIGGLECHAGTILSVDWSTDGDFILSCGFDHQLMEWDLSVKQVKEHLERACKALHQDKINVLTQSQDIPYVSKGTMRKSAVSRNIPDKEEDQLLELHRELIPRPSCLLPIYTPSSVSTDMHSDYVDCIRFLIGTNYALSKGCGNEKAIHFWRFGPPKGEVENRIHGNVLRPKSCTTKFRTMNVPSGSAWFIKFAVDPRRRWLVCGGAGGSVMFFDLRNNEETNPTHTCSVGSRTVRQASFSTCGRFLVLVTDEGFVCRFDRVSASVDAKDLAKF 0524A674ABAB2671 mes_2-mes_3-1 3 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait association association Fig 1a, 1B The anti-MES-2 antibodies were crosslinked to Protein A-agarose beads. mes_6-mes_2-1 3 6 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey association association Fig 1C mes_2-mes_6-1 1 6 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 4 unspecified role unspecified role bait bait 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 direct interaction direct interaction Fig. 2A Radiolabeled MES-6 was also precipitated by anti-MES-2 in the presence of cold MES-2 mes_6-mes_2-2 1 6 unspecified role unspecified role bait bait 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 4 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro direct interaction direct interaction Fig. 3B mes_2-mes_6-2 3 6 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association The interaction between MES-2 and MES-6 is stable under high salt conditions. Participants were identified by a mix of 2 antibodies, and by molecular weight separation. Fig. 4 mes_2-mes_6-3 2 5 unspecified role unspecified role neutral component neutral component 4 unspecified role unspecified role neutral component neutral component physical association physical association Mes-6 is assumed, but not demonstrated, to be present. Fig. 5