UniProt UniProt http://www.uniprot.org YPI1 and SDS22 proteins regulate the nuclear localization and function of yeast type 1 phosphatase Glc7. YPI1 and SDS22 proteins regulate the nuclear localization and function of yeast type 1 phosphatase Glc7. The Journal of biological chemistry 2007 imex curation Pedelini L., Marquina M., Ariño J., Casamayor A., Sanz L., Bollen M., Sanz P., Garcia-Gimeno MA. sanz@ibv.csic.es 15-Nov-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Pedelini L., Marquina M., Ariño J., Casamayor A., Sanz L., Bollen M., Sanz P., Garcia-Gimeno MA. sanz@ibv.csic.es Accepted 2017-SEP-04 AT 10:39 BST AT 10:39 BST by MGT The Journal of biological chemistry 2007 imex curation YPI1 and SDS22 proteins regulate the nuclear localization and function of yeast type 1 phosphatase Glc7. YPI1 and SDS22 proteins regulate the nuclear localization and function of yeast type 1 phosphatase Glc7. The Journal of biological chemistry 2007 imex curation Pedelini L., Marquina M., Ariño J., Casamayor A., Sanz L., Bollen M., Sanz P., Garcia-Gimeno MA. sanz@ibv.csic.es 15-Nov-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast phosphatase assay phosphatase assay predetermined predetermined participant predetermined Pedelini L., Marquina M., Ariño J., Casamayor A., Sanz L., Bollen M., Sanz P., Garcia-Gimeno MA. sanz@ibv.csic.es Accepted 2017-SEP-04 AT 10:39 BST AT 10:39 BST by MGT The Journal of biological chemistry 2007 imex curation YPI1 and SDS22 proteins regulate the nuclear localization and function of yeast type 1 phosphatase Glc7. YPI1 and SDS22 proteins regulate the nuclear localization and function of yeast type 1 phosphatase Glc7. The Journal of biological chemistry 2007 imex curation Pedelini L., Marquina M., Ariño J., Casamayor A., Sanz L., Bollen M., Sanz P., Garcia-Gimeno MA. sanz@ibv.csic.es 15-Nov-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Pedelini L., Marquina M., Ariño J., Casamayor A., Sanz L., Bollen M., Sanz P., Garcia-Gimeno MA. sanz@ibv.csic.es Accepted 2017-SEP-04 AT 10:39 BST AT 10:39 BST by MGT The Journal of biological chemistry 2007 imex curation YPI1 and SDS22 proteins regulate the nuclear localization and function of yeast type 1 phosphatase Glc7. YPI1 and SDS22 proteins regulate the nuclear localization and function of yeast type 1 phosphatase Glc7. The Journal of biological chemistry 2007 imex curation Pedelini L., Marquina M., Ariño J., Casamayor A., Sanz L., Bollen M., Sanz P., Garcia-Gimeno MA. sanz@ibv.csic.es 15-Nov-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column anti tag western anti tag western blot anti tag western Pedelini L., Marquina M., Ariño J., Casamayor A., Sanz L., Bollen M., Sanz P., Garcia-Gimeno MA. FPLC Superdex 200 HR 10/30 gel filtration column. HA-tagged proteins were further identified by molecular weight. sanz@ibv.csic.es Accepted 2017-SEP-04 AT 10:39 BST AT 10:39 BST by MGT The Journal of biological chemistry 2007 imex curation 4-nitrophenyl dihydrogen phosphate 4-nitrophenyl dihydrogen phosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) pp12_yeast Serine/threonine-protein phosphatase PP1-2 GLC7 CID1 DIS2 YER133W protein protein yeast Saccharomyces cerevisiae Baker's yeast MDSQPVDVDNIIDRLLEVRGSKPGQQVDLEENEIRYLCSKARSIFIKQPILLELEAPIKICGDIHGQYYDLLRLFEYGGFPPESNYLFLGDYVDRGKQSLETICLLLAYKIKYPENFFILRGNHECASINRIYGFYDECKRRYNIKLWKTFTDCFNCLPIAAIIDEKIFCMHGGLSPDLNSMEQIRRVMRPTDIPDVGLLCDLLWSDPDKDIVGWSENDRGVSFTFGPDVVNRFLQKQDMELICRAHQVVEDGYEFFSKRQLVTLFSAPNYCGEFDNAGAMMSVDESLLCSFQILKPAQKSLPRQAGGRKKK 71B20E3A4ED93235 71B20E3A4ED93235 sds22_yeast Protein phosphatase 1 regulatory subunit SDS22 YKL193C SDS22 EGP1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MDKNSVNKDSEEKDERHKIEVVDDTNPDFITADSELTQDLPDDVEVIDLVHLKIKSLEDLNLYRFKNLKQLCLRQNLIESISEVEVLPHDKIVDLDFYDNKIKHISSNVNKLTKLTSLDLSFNKIKHIKNLENLTDLENLYFVQNSISKIENLSTLKSLKNLELGGNKVHSIEPDSFEGLSNLEEIWLGKNSIPRLINLHPLKNLKILSIQSNKLKKIENLEELTNLEELYLSHNFITKIEGLEKNLKLTTLDVTSNKITSLENLNHLSNLTDIWASFNKIDQSFESLGENLSALSRLETIYLEGNPIQLENKTSYRRKLTMNLPPSLQKIDATYIRG 1EF434C7276ABAEA 1EF434C7276ABAEA ypi1_yeast Type 1 phosphatases regulator YPI1 YFR003C YPI1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSGNQMAMGSEQQQTVGSRTVSVEEVPAVLQLRATQDPPRSQEAMPTRHNVRWEENVIDNENMNKKKTKICCIFHPQNEDEEECNHHSDDDGSSSSGSSSSESENEKDLDFNERRQRRLERRHRKLEKKRSYSPNAYEIQPDYSEYRRKQQEKKD E191184DCD5EF8A1 E191184DCD5EF8A1 sds22-glc7-3 1 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 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 8 unspecified role unspecified role prey prey region lexa dna bd lexa dna binding domain lexa dna bd undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig. 1A ypi1-glc7-1 1 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 7 unspecified role unspecified role prey prey 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 8 unspecified role unspecified role bait bait region lexa dna bd lexa dna binding domain lexa dna bd undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig. 1B We first confirmed the interaction between Sds22 and Ypi1 by a direct co-immunoprecipitation method using cell extracts from yeast expressing GST-Sds22, LexA-Ypi1, and HA-Glc7. glc7-sds22-8 1 6 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey 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 8 unspecified role unspecified role prey prey region lexa dna bd lexa dna binding domain lexa dna bd undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig. 1C We first confirmed the interaction between Sds22 and Ypi1 by a direct co-immunoprecipitation method using cell extracts from yeast expressing GST-Sds22, LexA-Ypi1, and HA-Glc7. sds22-glc7-5 3 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Table 3, Fig. 3C sds22-glc7-4 3 8 unspecified role unspecified role ancillary ancillary region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Table 3 glc7-sds22-9 3 8 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role ancillary ancillary 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 Table 3 sds22-glc7-6 4 8 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 6 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 7 unspecified role unspecified role neutral component neutral component region lexa dna bd lexa dna binding domain lexa dna bd undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Table 3 sds22-ypi1-1 3 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Table 3, Fig 3B sds22-ypi1-2 3 8 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role prey prey physical association physical association Fig 3E ypi1-glc7-2 3 6 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Table 3, Fig. 3F 4_nitrophenyl dihydrogen phosphate-glc7-1 2 6 enzyme enzyme neutral component neutral component 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 enzyme target enzyme target substrate substrate neutral component neutral component dephosphorylation dephosphorylation reaction dephosphorylation Fig. 5 4_nitrophenyl dihydrogen phosphate-glc7-2 2 8 enzyme regulator enzyme regulator neutral component neutral component 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) 6 enzyme enzyme neutral component neutral component 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 enzyme target enzyme target substrate substrate neutral component neutral component dephosphorylation dephosphorylation reaction dephosphorylation Fig. 5 Because Ypi1 inhibits the phosphatase activity of Glc7 (Fig. 5) Ypi1 inhibits the phosphatase activity of Glc7 4_nitrophenyl dihydrogen phosphate-glc7-3 2 7 enzyme regulator enzyme regulator neutral component neutral component 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 6 enzyme enzyme neutral component neutral component 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 enzyme target enzyme target substrate substrate neutral component neutral component dephosphorylation dephosphorylation reaction dephosphorylation Fig. 5 As shown in Fig. 5, a GST-Sds22 fusion protein produced in yeast also inhibited (53% inhibition) the activity of Glc7 used in the assay. SDS22 inhibits the phosphatase activity of Glc7 4_nitrophenyl dihydrogen phosphate-glc7-4 2 8 enzyme regulator enzyme regulator neutral component neutral component 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) 7 enzyme regulator enzyme regulator neutral component neutral component 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 6 enzyme enzyme neutral component neutral component 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 enzyme target enzyme target substrate substrate neutral component neutral component dephosphorylation dephosphorylation reaction dephosphorylation Fig. 5 More interestingly, the combination of equimolar amounts of Ypi1 and Sds22 (0.224 μm each) displayed a higher inhibitory capacity on Glc7, leading to an almost full inhibition Ypi1 and Sds22 display an additive inhibition of the phosphatase activity of Glc7