UniProt UniProt http://www.uniprot.org Essential functions of Sds22p in chromosome stability and nuclear localization of PP1. Essential functions of Sds22p in chromosome stability and nuclear localization of PP1. Journal of cell science 2002 imex curation Peggie MW., MacKelvie SH., Bloecher A., Knatko EV., Tatchell K., Stark MJ. m.j.r.stark@dundee.ac.uk 05-Sep-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom fingerprinting peptide massfingerprinting fingerprinting Peggie MW., MacKelvie SH., Bloecher A., Knatko EV., Tatchell K., Stark MJ. The protein complex was purified on an IgG-Sepharose column then eluted following TEV cleavage of the Protein A tafg. m.j.r.stark@dundee.ac.uk Accepted 2017-SEP-01 AT 11:07 BST AT 11:07 BST by MGT Journal of cell science 2002 imex curation Essential functions of Sds22p in chromosome stability and nuclear localization of PP1. Essential functions of Sds22p in chromosome stability and nuclear localization of PP1. Journal of cell science 2002 imex curation Peggie MW., MacKelvie SH., Bloecher A., Knatko EV., Tatchell K., Stark MJ. m.j.r.stark@dundee.ac.uk 05-Sep-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast fluorescence imaging fluorescence microscopy fluorescence imaging anti-tag immunocytochemistry anti-tag immunocytochemistry Peggie MW., MacKelvie SH., Bloecher A., Knatko EV., Tatchell K., Stark MJ. m.j.r.stark@dundee.ac.uk Accepted 2017-SEP-01 AT 11:07 BST AT 11:07 BST by MGT Journal of cell science 2002 imex curation Essential functions of Sds22p in chromosome stability and nuclear localization of PP1. Essential functions of Sds22p in chromosome stability and nuclear localization of PP1. Journal of cell science 2002 imex curation Peggie MW., MacKelvie SH., Bloecher A., Knatko EV., Tatchell K., Stark MJ. m.j.r.stark@dundee.ac.uk 05-Sep-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 Peggie MW., MacKelvie SH., Bloecher A., Knatko EV., Tatchell K., Stark MJ. m.j.r.stark@dundee.ac.uk Accepted 2017-SEP-01 AT 11:07 BST AT 11:07 BST by MGT Journal of cell science 2002 imex curation 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 glc7-sds22-5 2 4 unspecified role unspecified role neutral component neutral component n-terminal 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 neutral component neutral component 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 colocalization colocalization Fig. 1 Although Sds22p largely co-localized with nuclear Glc7p glc7-sds22-6 1 4 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait c-terminal protein a tag protein a tag protein A protein A c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Fig. 2A, 2B However, comparison with a control strain expressing unfused Protein A showed that only two specific bands were present in the Sds22p-Protein A TEV eluate, and mass fingerprint analysis revealed that these were Sds22p and Glc7p Higher molecular weight bands were also observed but not identified. glc7-sds22-7 3 4 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role bait bait 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 P36047:p.[Ile54Thr;Ile193Thr] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain I T certain certain sequence position certain certain certain sequence position certain I T Wed May 02 14:57:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile54thr,ile193thr' Tue Jun 30 18:47:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ile54Thr;Ile193Thr]' physical association physical association Fig. 6 We therefore next tested by co-immunoprecipitation whether the formation of the Sds22p-Glc7p complex was affected by sds22 mutations. The SDS22 Ile54Thr,Ile1932Thr mutations reduced the interaction at 4C, and disrupted it at 30C. The interaction was conformed by immuno-depletion.