UniProt UniProt http://www.uniprot.org Assembly of scaffold-mediated complexes containing Cdc42p, the exchange factor Cdc24p, and the effector Cla4p required for cell cycle-regulated phosphorylation of Cdc24p. Assembly of scaffold-mediated complexes containing Cdc42p, the exchange factor Cdc24p, and the effector Cla4p required for cell cycle-regulated phosphorylation of Cdc24p. The Journal of biological chemistry 2001 imex curation Bose I., Irazoqui JE., Moskow JJ., Bardes ES., Zyla TR., Lew DJ. daniel.lew@duke.edu 17-May-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Bose I., Irazoqui JE., Moskow JJ., Bardes ES., Zyla TR., Lew DJ. daniel.lew@duke.edu Accepted 2018-MAY-09 AT 11:05 BST AT 11:05 BST by LIVIAP The Journal of biological chemistry 2001 Only protein-protein interactions imex curation imex curation Assembly of scaffold-mediated complexes containing Cdc42p, the exchange factor Cdc24p, and the effector Cla4p required for cell cycle-regulated phosphorylation of Cdc24p. Assembly of scaffold-mediated complexes containing Cdc42p, the exchange factor Cdc24p, and the effector Cla4p required for cell cycle-regulated phosphorylation of Cdc24p. The Journal of biological chemistry 2001 imex curation Bose I., Irazoqui JE., Moskow JJ., Bardes ES., Zyla TR., Lew DJ. daniel.lew@duke.edu 17-May-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Bose I., Irazoqui JE., Moskow JJ., Bardes ES., Zyla TR., Lew DJ. daniel.lew@duke.edu Accepted 2018-MAY-09 AT 11:05 BST AT 11:05 BST by LIVIAP The Journal of biological chemistry 2001 Only protein-protein interactions imex curation imex curation Assembly of scaffold-mediated complexes containing Cdc42p, the exchange factor Cdc24p, and the effector Cla4p required for cell cycle-regulated phosphorylation of Cdc24p. Assembly of scaffold-mediated complexes containing Cdc42p, the exchange factor Cdc24p, and the effector Cla4p required for cell cycle-regulated phosphorylation of Cdc24p. The Journal of biological chemistry 2001 imex curation Bose I., Irazoqui JE., Moskow JJ., Bardes ES., Zyla TR., Lew DJ. daniel.lew@duke.edu 17-May-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Bose I., Irazoqui JE., Moskow JJ., Bardes ES., Zyla TR., Lew DJ. daniel.lew@duke.edu Accepted 2018-MAY-09 AT 11:05 BST AT 11:05 BST by LIVIAP The Journal of biological chemistry 2001 Only protein-protein interactions imex curation imex curation cdc24_yeast Cell division control protein 24 CLS4 YAL041W CDC24 Calcium regulatory protein protein protein yeast Saccharomyces cerevisiae Baker's yeast MAIQTRFASGTSLSDLKPKPSATSISIPMQNVMNKPVTEQDSLFHICANIRKRLEVLPQLKPFLQLAYQSSEVLSERQSLLLSQKQHQELLKSNGANRDSSDLAPTLRSSSISTATSLMSMEGISYTNSNPSATPNMEDTLLTFSMGILPITMDCDPVTQLSQLFQQGAPLCILFNSVKPQFKLPVIASDDLKVCKKSIYDFILGCKKHFAFNDEELFTISDVFANSTSQLVKVLEVVETLMNSSPTIFPSKSKTQQIMNAENQHRHQPQQSSKKHNEYVKIIKEFVATERKYVHDLEILDKYRQQLLDSNLITSEELYMLFPNLGDAIDFQRRFLISLEINALVEPSKQRIGALFMHSKHFFKLYEPWSIGQNAAIEFLSSTLHKMRVDESQRFIINNKLELQSFLYKPVQRLCRYPLLVKELLAESSDDNNTKELEAALDISKNIARSINENQRRTENHQVVKKLYGRVVNWKGYRISKFGELLYFDKVFISTTNSSSEPEREFEVYLFEKIIILFSEVVTKKSASSLILKKKSSTSASISASNITDNNGSPHHSYHKRHSNSSSSNNIHLSSSSAAAIIHSSTNSSDNNSNNSSSSSLFKLSANEPKLDLRGRIMIMNLNQIIPQNNRSLNITWESIKEQGNFLLKFKNEETRDNWSSCLQQLIHDLKNEQFKARHHSSTSTTSSTAKSSSMMSPTTTMNTPNHHNSRQTHDSMASFSSSHMKRVSDVLPKRRTTSSSFESEIKSISENFKNSIPESSILFRISYNNNSNNTSSSEIFTLLVEKVWNFDDLIMAINSKISNTHNNNISPITKIKYQDEDGDFVVLGSDEDWNVAKEMLAENNEKFLNIRLY 856BCAF4EB0A67D1 856BCAF4EB0A67D1 bem1_yeast Bud emergence protein 1 Suppressor of RHO3 protein 1 BEM1 SRO1 YBR200W YBR1412 protein protein yeast Saccharomyces cerevisiae Baker's yeast MLKNFKLSKRDSNGSKGRITSADISTPSHDNGSVIKHIKTVPVRYLSSSSTPVKSQRDSSPKNRHNSKDITSPEKVIKAKYSYQAQTSKELSFMEGEFFYVSGDEKDWYKASNPSTGKEGVVPKTYFEVFDRTKPSSVNGSNSSSRKVTNDSLNMGSLYAIVLYDFKAEKADELTTYVGENLFICAHHNCEWFIAKPIGRLGGPGLVPVGFVSIIDIATGYATGNDVIEDIKSVNLPTVQEWKSNIARYKASNISLGSVEQQQQQSITKPQNKSAKLVDGELLVKASVESFGLEDEKYWFLVCCELSNGKTRQLKRYYQDFYDLQVQLLDAFPAEAGKLRDAGGQWSKRIMPYIPGPVPYVTNSITKKRKEDLNIYVADLVNLPDYISRSEMVHSLFVVLNNGFDREFERDENQNNIKTLQENDTATFATASQTSNFASTNQDNTLTGEDLKLNKKLSDLSLSGSKQAPAQSTSGLKTTKIKFYYKDDIFALMLKGDTTYKELRSKIAPRIDTDNFKLQTKLFDGSGEEIKTDSQVSNIIQAKLKISVHDI FB6C525AE9A19181 FB6C525AE9A19181 cdc42_yeast Cell division control protein 42 CDC42 SRO2 YLR229C L8083.13 Suppressor of RHO3 protein 2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MQTLKCVVVGDGAVGKTCLLISYTTNQFPADYVPTVFDNYAVTVMIGDEPYTLGLFDTAGQEDYDRLRPLSYPSTDVFLVCFSVISPPSFENVKEKWFPEVHHHCPGVPCLVVGTQIDLRDDKVIIEKLQRQRLRPITSEQGSRLARELKAVKYVECSALTQRGLKNVFDEAIVAALEPPVIKKSKKCAIL 30BD644BA9836B2C 30BD644BA9836B2C cla4_yeast Serine/threonine-protein kinase CLA4 CLA4 N0450 YNL298W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSLSAAANKISDNDFQNIGPAPRPPSSNSQGRTCYNQTQPITKLMSQLDLTSASHLGTSTSKKKSGWVSYKDDGILSFIWQKRYLMLHDSYVALYKNDKQNDDAILKIPLTSIISVSRTQLKQYCFELVRCSDRNSVSSGSSSSLNVSSDSNSKKSIYIATKTESDLHSWLDAIFAKCPLLSGVSSPTNFTHKVHVGFDPETGSFVGMPTNWEKLLKHSRITGEDWNNNSAAVIQVLQFYQEYNGAGNPTNTLDKPQSGETSSSQKSLPNSYNDNKLRNNSVNSKSSSGVSSSMVSQRKTSQPPNTKSPVSLGSGSLPPINTKLPTSQSNIPRHLQNVPNQQYPKMRNGHSPTNGQFPRGPMHPNNSQRSLQQQQQQQQQQKQQHQQYPYHHQGPSPSPSPSPSPLNPYRPHHNMINPYSKQPQSPLSSQSTQNQAIPRYAQNSSPTAAHFQPQRTAPKPPISAPRAPYPSNQNATSNTHVQPVAPKNDQSTPQTMRQAPKRPDADVAQPGGVAKPKKPARPTMSTAEIMSKLKKVTVNADPSQCFKVIEKAGQGASGSVYLAERTHIPTESNMIELINNDIDEPHVGDKVAIKQMVLSKQPRKELIVNEILVMKDSRHKNIVNFLEAYLRTDDDLWVVMEFMEGGSLTDIIENSPTNDNSHSPLTEPQIAYIVRETCQGLKFLHDKHIIHRDIKSDNVLLDTRARVKITDFGFCARLTDKRSKRATMVGTPYWMAPEVVKQREYDEKIDVWSLGIMTIEMLEGEPPYLNEDPLKALYLIATNGTPKLKHPESLSLEIKRFLSVCLCVDVRYRASTEELLHHGFFNMACDPKDLTSLLEWKE 3A2AB86414561DC6 cdc42-bem1-1 3 6 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell c-terminal 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 physical association physical association Specifically, we found that Myc-tagged Bem1p associated with FLAG-tagged Cdc42p (Fig. 3A) Fig. 3A bem1-cla4-1 1 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate n-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range yeast Saccharomyces cerevisiae Baker's yeast 5 unspecified role unspecified role bait bait purified purified n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Fig. 4D, Fig 5D Myc-tagged Cla4p bound to GST-Bem1p (but not GST alone; see Fig. 4D). bem1-cdc24-2 3 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell c-terminal 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 physical association physical association Fig. 3C, 3F The amount of CDC24 associated with BEM1 was unaffected regardless of whether wild-type, GTP-locked, or GDP-locked CDC42 was expressed in the cells. The ohosphorylation state of CDC24 also did not affect BEM1 binding. cla4-cdc24-1 3 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 3D, 3F The amount of CDC24 associated with CLA4 was unaffected regardless of whether wild-type, GTP-locked, or GDP-locked Cdc42p was expressed in the cells. In addition, HA-tagged Cdc24p associated with Myc-tagged Cla4p (Fig. 3D) cdc42-cdc24-1 3 6 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Specifically, we found that Myc-tagged Bem1p associated with FLAG-tagged Cdc42p (Fig. 3A) Fig. 3E cdc42-bem1-2 1 6 unspecified role unspecified role bait bait purified purified P19073:p.Gln61Leu mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain Q L Tue Jun 30 17:07:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln61Leu' n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range P19073:p.Asp57Tyr mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain D Y Wed May 02 18:03:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp57tyr' Tue Jun 30 19:12:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp57Tyr' ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate c-terminal 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 region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain yeast Saccharomyces cerevisiae Baker's yeast physical association physical association Myc-tagged Bem1p bound efficiently to GST-Cdc42p Q61L, but not GST-Cdc42p D57Y, in vitro (Fig. 4A). Fig. 4A, 4E, 5A The amount of CDC24 associated with BEM1 was unaffected regardless of whether wild-type, GTP-locked, or GDP-locked CDC42 was expressed in the cells. The ohosphorylation state of CDC24 also did not affect BEM1 binding. bem1-cdc42-1 1 6 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate P19073:p.Asp57Tyr mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain D Y Wed May 02 15:59:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp57tyr' Tue Jun 30 20:03:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp57Tyr' P19073:p.Gln61Leu mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 16:38:41 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln61leu' Tue Jun 30 17:38:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln61Leu' c-terminal 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 ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role bait bait purified purified n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Fig. 4B, 5D The amount of CDC24 associated with BEM1 was unaffected regardless of whether wild-type, GTP-locked, or GDP-locked CDC42 was expressed in the cells. The ohosphorylation state of CDC24 also did not affect BEM1 binding. cla4-bem1-3 1 7 unspecified role unspecified role bait bait n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate c-terminal 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 region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain yeast Saccharomyces cerevisiae Baker's yeast physical association physical association Fig. 4C, 4E In addition, Myc-tagged Bem1p bound to GST-Cla4p (but not GST alone; see Fig. 4C) cdc24-bem1-6 1 6 weight identificat confirmation by molecular weight weight identificat anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate P19073:p.Gln61Leu mutation mutation certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 15:25:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln61leu' Tue Jun 30 19:18:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln61Leu' region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast 4 unspecified role unspecified role bait bait purified purified 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 ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate c-terminal 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 yeast Saccharomyces cerevisiae Baker's yeast association association Fig. 5A cdc24-bem1-7 1 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate n-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range yeast Saccharomyces cerevisiae Baker's yeast 4 unspecified role unspecified role bait bait purified purified 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 ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate c-terminal 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 yeast Saccharomyces cerevisiae Baker's yeast association association Fig. 5A The amount of CDC24 associated with BEM1 was unaffected regardless of whether wild-type, GTP-locked, or GDP-locked CDC42 was expressed in the cells. The ohosphorylation state of CDC24 also did not affect BEM1 binding. cdc24-bem1-8 1 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate n-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range yeast Saccharomyces cerevisiae Baker's yeast 6 weight identificat confirmation by molecular weight weight identificat anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate P19073:p.Gln61Leu mutation mutation certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 14:52:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln61leu' Tue Jun 30 15:54:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln61Leu' region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast 4 unspecified role unspecified role bait bait purified purified 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 ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate c-terminal 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 yeast Saccharomyces cerevisiae Baker's yeast association association Fig. 5A bem1-cdc24-6 1 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate n-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range yeast Saccharomyces cerevisiae Baker's yeast 5 unspecified role unspecified role bait bait purified purified n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Fig. 5C, 5D bem1-cdc24-7 2 4 enzyme target enzyme target substrate substrate prey prey cell lysate cell lysate region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast 7 enzyme enzyme ancillary ancillary cell lysate cell lysate n-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range yeast Saccharomyces cerevisiae Baker's yeast 5 enzyme regulator enzyme regulator bait bait purified purified n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Fig. 5C, 5D Incubation of these Cla4p-Bem1p-Cdc24p complexes with [g-32P]ATP promoted 32P incorporation into the Myc-Cdc24p but not into the Myc-Cla4p (Fig. 5C). Phosphorylation of CDC24 was unaffected by the presence or absence of CDC42 bem1-cdc24-8 1 6 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate n-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range yeast Saccharomyces cerevisiae Baker's yeast 5 unspecified role unspecified role bait bait purified purified n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) association association Fig. 5D bem1-cla4-2 3 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell c-terminal 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 physical association physical association Fig. 3B The amount of CDC24 associated with BEM1 was unaffected regardless of whether wild-type, GTP-locked, or GDP-locked CDC42 was expressed in the cells. The ohosphorylation state of CDC24 also did not affect BEM1 binding. bem1-cdc24-5 1 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate c-terminal 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 ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role bait bait purified purified n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Fig. 5D The amount of CDC24 associated with BEM1 was unaffected regardless of whether wild-type, GTP-locked, or GDP-locked CDC42 was expressed in the cells. The ohosphorylation state of CDC24 also did not affect BEM1 binding.