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/ http://www.ebi.ac.uk/ chang-2005-1 Interaction of checkpoint proteins Hus1/Rad1/Rad9 with DNA base excision repair enzyme MutY homolog in fission yeast, Schizosaccharomyces pombe. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Chang DY., Lu AL. An S.pombe strain expressing a recombinant Hemagglutin (HA) tagged Hus1 and Rad1 proteins was used in this study. Figure 1. Figure 2. alchang@umaryland.edu J. Biol. Chem. (0021-9258) 2005 chang-2005-2 Interaction of checkpoint proteins Hus1/Rad1/Rad9 with DNA base excision repair enzyme MutY homolog in fission yeast, Schizosaccharomyces pombe. schpm Schizosaccharomyces pombe (fission yeast) Fission yeast anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Chang DY., Lu AL. An S.pombe strain expressing a recombinant Hemagglutin (HA) tagged Hus1 and Rad1 proteins was used in this study. Figure 1. Figure 2. alchang@umaryland.edu J. Biol. Chem. (0021-9258) 2005 chang-2005-3 Interaction of checkpoint proteins Hus1/Rad1/Rad9 with DNA base excision repair enzyme MutY homolog in fission yeast, Schizosaccharomyces pombe. schpm Schizosaccharomyces pombe (fission yeast) Fission yeast molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Chang DY., Lu AL. An S.pombe strain expressing a recombinant Hemagglutin (HA) tagged Hus1 and Rad1 proteins was used in this study. Figure 1. Figure 2. alchang@umaryland.edu J. Biol. Chem. (0021-9258) 2005 hus1_schpo Checkpoint protein hus1 hus1 SPAC20G4.04c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MRFKTRISNLYTLTRLVQALDKIGRFCWLRLMPETVNFVIVPDFRMTQVWSVLEVETIFEDYVVQSNADNVINLEVPIDNFYKALRSAANASDSTVRLSKKNNQPLLSLSTTWSGRAFGSNIVTHNIPVRVLSQSYVSVIKEPTAPEPDCHIFLPQLNFLRHVVDKYKSLSDRIIMSANMSGELQLSVNIPSARVSTKWKGLENPELDPSQVEDISRHPSQTRAPEEFVHMRLDSKDLVNMLKISSVAKRVIACFCEGHALVLYVYITDPEDEHTAVLTYYISTYVD myh1_schpo Adenine DNA glycosylase myh1 myh SPAC26A3.02 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSDSNHSLDLHSYTQLEVERFRESLIQFYDKTKRILPWRKKECIPPSEDSPLEDWEQPVQRLYEVLVSEIMLQQTRVETVKRYYTKWMETLPTLKSCAEAEYNTQVMPLWSGMGFYTRCKRLHQACQHLAKLHPSEIPRTGDEWAKGIPGVGPYTAGAVLSIAWKQPTGIVDGNVIRVLSRALAIHSDCSKGKANALIWKLANELVDPVRPGDFNQALMELGAITCTPQSPRCSVCPISEICKAYQEQNVIRDGNTIKYDIEDVPCNICITDIPSKEDLQNWVVARYPVHPAKTKQREERALVVIFQKTDPSTKEKFFLIRKRPSAGLLAGLWDFPTIEFGQESWPKDMDAEFQKSIAQWISNDSRSLIKKYQSRGRYLHIFSHIRKTSHVFYAIASPDIVTNEDFFWISQSDLEHVGMCELGLKNYRAALEIKKRKVTSLSNFKEPKLTSARRIVTKAEC rad1_schpo DNA damage checkpoint control protein rad1 rad1 SPAC1952.07 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MFQAETVCLKQIQSTLRCIDFSKECTIEITSRGLRFAVEESQSLQAHAFLDKSLFQTFNFQGDSDGDTYMFQTMISPLLQSLSIYTDGKERISTSAWDQPTVNIMHKRGVICKVQYNGPGCPFIWEVEEMAGYATACELLTMECEDDVDINRLASTLCTKIIMKSNWLYDALVELDNNMGENLIIHTSSQKSTFLLRCVGALSTTEIEYPNEKSVLESFETDSENTYSYRFSLIRHALKALQVGSKVNLRIDENGTLSIQIMLVGQEGLCTFVDFCIVPLDLVSEDEEEDEEEEPAESNQSDNNVLRNDPNYRGDAETEDEDS rad9_schpo DNA repair protein rad9 rad9 SPAC664.07c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MEFTVSNVNLRDLARIFTNLSRIDDAVNWEINKNQIEITCLNSSRSGFSMVTLKKAFFDKYIFQPDSVLLTGLMTPTIRIRTQVKPILSVFRNKIFDFIPTVVTTNSKNGYGSESASRKDVIVENVQISISTGSECRIIFKFLCKHGVIKTYKISYEQTQTLHAVFDKSLSHNNFQINSKILKDLTEHFGQRTEELTIQPLQERVLLTSFTEEVVHNRDILKQPTQTTVSIDGKEFERVALNEGVSVTLSLREFRAAVILAEALGSSICAYYGVPGKPILLTFAKGKNSEIEAQFILATVVGSDEQEVSSMMGNRWQHSSTPASLFNSVERNNSLTAVAHNPPGSIGWQTDQSDSSRMFNSALDRSDETNGIKEPSTTNDAGQSLFLDGIPNESELAAFNNDVNDDAEFGPTQAEQSYHGIFSQED pcna_schpo Proliferating cell nuclear antigen pcn1 pcn SPBC16D10.09 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MLEARFQQAALLKKLLDAIKELVTDANFDCNDNGISLQAMDSSHVALVSMLIKSDGFEPYRCDRNIALGINLNALSKVLRCAQNEDLVTLKAEDTPEVLNLVFESEKNDRISDYDVKLMDIDQEHLGIPDIEYDATITMPAAEFQRITRDLLTLSDSVTINASKEGVRFSCKGDIGNGSTTLKQHTDLSDQDQSIEISLTQAVTLTFSLKYLAQFTKATPLATRVTLSMSNDVPLLVEYKMESGFLRFYLAPKIGEEDEE 1EE5B2A27A899C4E myh1-hus1 GST pulldown analysis reveals interaction between the MYH protein and the HUS1 protein 258464 N/A 258467 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 258468 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobb Escherichia coli (strain B / BL21) physical association physical association false false false Authors note that the results of this study indicate that HUS1 binds MYH within a central region of the protein or that a major binding region was located within residues 245 to 461 amino acids of the MYH protein. While a minor binding region was located within residues 1 to 230 amino acids of MYH. hus1-myh1 His pulldown analysis reveals interaction between the MYH protein and the HUS1 protein 258464 N/A 258467 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 false ecobb Escherichia coli (strain B / BL21) N/A 258468 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) physical association physical association false false false myh1-rad1 GST pulldown analysis reveals interaction between the MYH protein and the Rad1 protein 258464 N/A 258469 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 258468 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobb Escherichia coli (strain B / BL21) physical association physical association false false false Authors state that the major binding site for Rad1 is within the C-terminal (245aa - 323aa) region of the MYH protein. myh1-rad1-1 GST pulldown analysis reveals interaction between the MYH protein and the Rad1 protein 258464 N/A 258469 unspecified role unspecified role prey prey 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 false ecobb Escherichia coli (strain B / BL21) N/A 258468 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false myh1-rad9 GST pulldown analysis reveals interaction between the MYH protein and the Rad9 protein 258464 N/A 258468 unspecified role unspecified role bait bait region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobb Escherichia coli (strain B / BL21) N/A 258470 unspecified role unspecified role prey prey 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 false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false myh1-hus1-1 Coimmunoprecipitation analysis reveals interaction between the MYH protein and the HUS1 protein is stimulated by oxidative stress induction 258465 N/A 258467 unspecified role unspecified role prey prey 13myc-tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 258468 unspecified role unspecified role bait bait schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association false false false Hydrogen peroxide induced oxidative stress Hus1 and Myh1 interaction was found to be greatest 2 hours after hydrogen peroxide treatment. myh1-pcn1 Coimmunoprecipitation analysis reveals interaction between the MYH protein and the PCN1 protein is stimulated by oxidative stress induction 258465 N/A 258468 unspecified role unspecified role bait bait N/A 258471 unspecified role unspecified role prey prey physical association physical association false false false myh1-hus1-2 gel filtration analysis reveals interaction between the MYH protein and the HUS1 protein 258466 N/A 258467 unspecified role unspecified role neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 258468 unspecified role unspecified role neutral component neutral component region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association false false false MYH1 and Hus1 associate in a complex of approximately 450kDa