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/ Interaction of checkpoint proteins Hus1/Rad1/Rad9 with DNA base excision repair enzyme MutY homolog in fission yeast, Schizosaccharomyces pombe. Interaction of checkpoint proteins Hus1/Rad1/Rad9 with DNA base excision repair enzyme MutY homolog in fission yeast, Schizosaccharomyces pombe. J. Biol. Chem. (0021-9258) 2005 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 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 Annotation to indicate that this entry has been checked and accepted by a senior curator. J. Biol. Chem. (0021-9258) 2005 Interaction of checkpoint proteins Hus1/Rad1/Rad9 with DNA base excision repair enzyme MutY homolog in fission yeast, Schizosaccharomyces pombe. Interaction of checkpoint proteins Hus1/Rad1/Rad9 with DNA base excision repair enzyme MutY homolog in fission yeast, Schizosaccharomyces pombe. J. Biol. Chem. (0021-9258) 2005 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 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 Annotation to indicate that this entry has been checked and accepted by a senior curator. J. Biol. Chem. (0021-9258) 2005 Interaction of checkpoint proteins Hus1/Rad1/Rad9 with DNA base excision repair enzyme MutY homolog in fission yeast, Schizosaccharomyces pombe. Interaction of checkpoint proteins Hus1/Rad1/Rad9 with DNA base excision repair enzyme MutY homolog in fission yeast, Schizosaccharomyces pombe. J. Biol. Chem. (0021-9258) 2005 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 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 Annotation to indicate that this entry has been checked and accepted by a senior curator. J. Biol. Chem. (0021-9258) 2005 myh1_schpo Adenine DNA glycosylase myh1 myh SPAC26A3.02 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSDSNHSLDLHSYTQLEVERFRESLIQFYDKTKRILPWRKKECIPPSEDSPLEDWEQPVQRLYEVLVSEIMLQQTRVETVKRYYTKWMETLPTLKSCAEAEYNTQVMPLWSGMGFYTRCKRLHQACQHLAKLHPSEIPRTGDEWAKGIPGVGPYTAGAVLSIAWKQPTGIVDGNVIRVLSRALAIHSDCSKGKANALIWKLANELVDPVRPGDFNQALMELGAITCTPQSPRCSVCPISEICKAYQEQNVIRDGNTIKYDIEDVPCNICITDIPSKEDLQNWVVARYPVHPAKTKQREERALVVIFQKTDPSTKEKFFLIRKRPSAGLLAGLWDFPTIEFGQESWPKDMDAEFQKSIAQWISNDSRSLIKKYQSRGRYLHIFSHIRKTSHVFYAIASPDIVTNEDFFWISQSDLEHVGMCELGLKNYRAALEIKKRKVTSLSNFKEPKLTSARRIVTKAEC 5908E880202E2242 pcna_schpo Proliferating cell nuclear antigen pcn1 pcn SPBC16D10.09 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MLEARFQQAALLKKLLDAIKELVTDANFDCNDNGISLQAMDSSHVALVSMLIKSDGFEPYRCDRNIALGINLNALSKVLRCAQNEDLVTLKAEDTPEVLNLVFESEKNDRISDYDVKLMDIDQEHLGIPDIEYDATITMPAAEFQRITRDLLTLSDSVTINASKEGVRFSCKGDIGNGSTTLKQHTDLSDQDQSIEISLTQAVTLTFSLKYLAQFTKATPLATRVTLSMSNDVPLLVEYKMESGFLRFYLAPKIGEEDEE 1EE5B2A27A899C4E 1EE5B2A27A899C4E rad1_schpo DNA damage checkpoint control protein rad1 rad1 SPAC1952.07 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MFQAETVCLKQIQSTLRCIDFSKECTIEITSRGLRFAVEESQSLQAHAFLDKSLFQTFNFQGDSDGDTYMFQTMISPLLQSLSIYTDGKERISTSAWDQPTVNIMHKRGVICKVQYNGPGCPFIWEVEEMAGYATACELLTMECEDDVDINRLASTLCTKIIMKSNWLYDALVELDNNMGENLIIHTSSQKSTFLLRCVGALSTTEIEYPNEKSVLESFETDSENTYSYRFSLIRHALKALQVGSKVNLRIDENGTLSIQIMLVGQEGLCTFVDFCIVPLDLVSEDEEEDEEEEPAESNQSDNNVLRNDPNYRGDAETEDEDS 1A2A4DEFC3A55EE6 hus1_schpo Checkpoint protein hus1 hus1 SPAC20G4.04c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MRFKTRISNLYTLTRLVQALDKIGRFCWLRLMPETVNFVIVPDFRMTQVWSVLEVETIFEDYVVQSNADNVINLEVPIDNFYKALRSAANASDSTVRLSKKNNQPLLSLSTTWSGRAFGSNIVTHNIPVRVLSQSYVSVIKEPTAPEPDCHIFLPQLNFLRHVVDKYKSLSDRIIMSANMSGELQLSVNIPSARVSTKWKGLENPELDPSQVEDISRHPSQTRAPEEFVHMRLDSKDLVNMLKISSVAKRVIACFCEGHALVLYVYITDPEDEHTAVLTYYISTYVD 283CE8F538DEFB78 rad9_schpo DNA repair protein rad9 rad9 SPAC664.07c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MEFTVSNVNLRDLARIFTNLSRIDDAVNWEINKNQIEITCLNSSRSGFSMVTLKKAFFDKYIFQPDSVLLTGLMTPTIRIRTQVKPILSVFRNKIFDFIPTVVTTNSKNGYGSESASRKDVIVENVQISISTGSECRIIFKFLCKHGVIKTYKISYEQTQTLHAVFDKSLSHNNFQINSKILKDLTEHFGQRTEELTIQPLQERVLLTSFTEEVVHNRDILKQPTQTTVSIDGKEFERVALNEGVSVTLSLREFRAAVILAEALGSSICAYYGVPGKPILLTFAKGKNSEIEAQFILATVVGSDEQEVSSMMGNRWQHSSTPASLFNSVERNNSLTAVAHNPPGSIGWQTDQSDSSRMFNSALDRSDETNGIKEPSTTNDAGQSLFLDGIPNESELAAFNNDVNDDAEFGPTQAEQSYHGIFSQED A5C1173D43688E98 myh1-hus1 3 7 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 schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 4 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 ecobb Escherichia coli (strain B / BL21) physical association physical association 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 3 7 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 ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) physical association physical association myh1-rad1 3 6 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 schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 4 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 ecobb Escherichia coli (strain B / BL21) physical association physical association Authors state that the major binding site for Rad1 is within the C-terminal (245aa - 323aa) region of the MYH protein. myh1-rad1-1 3 6 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 ecobb Escherichia coli (strain B / BL21) 4 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 region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction myh1-rad9 3 4 unspecified role unspecified role bait bait region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) 8 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction myh1-hus1-1 2 7 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 schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 4 unspecified role unspecified role bait bait schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association Hydrogen peroxide induced oxidative stress Hus1 and Myh1 interaction was found to be greatest 2 hours after hydrogen peroxide treatment. myh1-pcn1 2 4 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey physical association physical association myh1-hus1-2 1 7 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 schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 4 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 schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association MYH1 and Hus1 associate in a complex of approximately 450kDa