UniProt UniProt http://www.uniprot.org Hsk1-Dfp1 is required for heterochromatin-mediated cohesion at centromeres. Hsk1-Dfp1 is required for heterochromatin-mediated cohesion at centromeres. Nat. Cell Biol. (1465-7392) 2003 Bailis JM., Bernard P., Antonelli R., Allshire RC., Forsburg SL. Chromatin - Epigenetic interactions resulting in chromatin modulation forsburg@salk.edu 15-NOV-2006: Contacted by JYOTI. in vitro In vitro kinase htrf kinase homogeneous time resolved fluorescence kinase HTRF homogeneous time-resolved fluorescence kinase htrf predetermined predetermined participant predetermined Bailis JM., Bernard P., Antonelli R., Allshire RC., Forsburg SL. Chromatin - Epigenetic interactions resulting in chromatin modulation revisit when interaction detection term resolved - this is a straight in vitro kinase assay. forsburg@salk.edu Accepted 2006-NOV-1 by JYOTI. 15-NOV-2006: Contacted by JYOTI. Nat. Cell Biol. (1465-7392) 2003 Hsk1-Dfp1 is required for heterochromatin-mediated cohesion at centromeres. Hsk1-Dfp1 is required for heterochromatin-mediated cohesion at centromeres. Nat. Cell Biol. (1465-7392) 2003 Bailis JM., Bernard P., Antonelli R., Allshire RC., Forsburg SL. Chromatin - Epigenetic interactions resulting in chromatin modulation forsburg@salk.edu 15-NOV-2006: Contacted by JYOTI. schpm Schizosaccharomyces pombe (fission yeast) Fission yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Bailis JM., Bernard P., Antonelli R., Allshire RC., Forsburg SL. Chromatin - Epigenetic interactions resulting in chromatin modulation forsburg@salk.edu Accepted 2006-NOV-1 by JYOTI. 15-NOV-2006: Contacted by JYOTI. Nat. Cell Biol. (1465-7392) 2003 Hsk1-Dfp1 is required for heterochromatin-mediated cohesion at centromeres. Hsk1-Dfp1 is required for heterochromatin-mediated cohesion at centromeres. Nat. Cell Biol. (1465-7392) 2003 Bailis JM., Bernard P., Antonelli R., Allshire RC., Forsburg SL. Chromatin - Epigenetic interactions resulting in chromatin modulation forsburg@salk.edu 15-NOV-2006: Contacted by JYOTI. schpm Schizosaccharomyces pombe (fission yeast) Fission yeast anti tag coip anti tag coimmunoprecipitation anti tag coip polyclonal western polyclonal antibody western blot polyclonal western Bailis JM., Bernard P., Antonelli R., Allshire RC., Forsburg SL. Chromatin - Epigenetic interactions resulting in chromatin modulation forsburg@salk.edu Accepted 2006-NOV-1 by JYOTI. 15-NOV-2006: Contacted by JYOTI. Nat. Cell Biol. (1465-7392) 2003 him1_schpo Hsk1-interacting molecule 1 SPCC550.13 him1 dfp1 rad35 DNA repair protein rad35 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MNLGRCPLAPRSANIVLPKHDAVSKQKEYRIEEKTNEAQREEIITWKDNREDEGEVKTDFEVVNNENIITTTPKHQTVITPKSYRKSVKRIKHDAPQNEDIPVMKGLAPINADTESKAESMAAGKVLGSKNSSQKARLQEWKRQYKKAFPHFRFYLDGCDPSVAHRVKKQIQQLGGHVETFFSGNVTHVATVRAIQDVSVKYAKQDVITKARQLNMKIWSMEKLCNRVLKTLMENDQCTTNAPTKQGNDLSYLLYVEKVQGTNERDLSVPRQDFVHFRGPYLYVHDIANIYKPILLREWQKPLPDRDVPWPTFRATSIGRCPFVPETKYRLSTSKSLVAKNDQQLLQRQSQEPSLILRANSMKASLPDISNTGISGMNTNTTYNTNINNTPQTAISGITQDTSPSIRTNCHHCLDDGMQASGIVQSNLTSAAMSNNSAIRSGSAASVPVVTINGRDIAELKKRIIQQKSGMIGKDYSYKAMLHNTSQRKIRVDAKPGYCENCREKFDNFESHIRSSRHRRFAENNDNFKDLDELFALVQRPLRPD 96F90243322AFD8D hsk1_schpo Cell cycle serine/threonine-protein kinase hsk1 hsk1 SPBC776.12c Minichromosome maintenance protein kinase Cdc7-related kinase protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MAEAHITLSPKVTHEQQTDIDSECEITEVDDENVNENKSQEMIQDIPARDREEIENITRTFVELQENYRLIEKIGEGTFSSVYKAEDLHYGRYINDWDIQSEVLKESSFGKEKIPVNEDSRKPKYVAIKKIYATSSPARIYNELEILYLLRGSSVIAPLITALRNEDQVLVVLPYYEHTDFRQYYSTFSYRDMSIYFRCLFQAMQQTQTLGIIHRDIKPSNFLFDVRTKHGVLVDFGLAERYDGRQQSHSCRCTNSNAAELAHDFSIAQETSLGYIKNDTRPSKRANRAGTRGFRAPEVLFKCSSQSPKVDIWSAGVILLSFLTKRFPMFNSKDDVDALMEIACIFGKSEMRQCAALHGCTFETNVSTLTEKRVNFRKLILWASCGSASIYKEKLRHKPSQEERLCLDFLEKCLELDCNKRISAEEALDHDFLYLDNLAYEKKDDDTAFDNSFGETSFEKDEDLTAKHLSHILDFKEQEETDEPTSLSKRKRSIDEILPNDALQDGA 04970E58218441EE swi6_schpo Chromatin-associated protein swi6 swi6 SPAC664.01c SPAC824.10c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MKKGGVRSYRRSSTSKRSVIDDDSEPELPSMTKEAIASHKADSGSSDNEVESDHESKSSSKKLKENAKEEEGGEEEEEDEYVVEKVLKHRMARKGGGYEYLLKWEGYDDPSDNTWSSEADCSGCKQLIEAYWNEHGGRPEPSKRKRTARPKKPEAKEPSPKSRKTDEDKHDKDSNEKIEDVNEKTIKFADKSQEEFNENGPPSGQPNGHIESDNESKSPSQKESNESEDIQIAETPSNVTPKKKPSPEVPKLPDNRELTVKQVENYDSWEDLVSSIDTIERKDDGTLEIYLTWKNGAISHHPSTITNKKCPQKMLQFYESHLTFRENE F505CC8AC5E75EAB F505CC8AC5E75EAB scc3_schpo Cohesin subunit psc3 SCC3 homolog psc3 SPAC17H9.20 SPAC607.01 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSESVTTGSDDDGGDRESSPVMLSQSFDPMSSSSNSSSEENSDDDYEKTISSKKRHPRPNSKGVNVKRSRRNAIVEEDPQEEIFNNLFAFLLDQKVDTMDIAVSWFADYAKDNQSALANLINFILKCCGCNRAINVFDVQDQDSASATLSQIQLSVERTSTRDYPLNSKNLKFRNFRKRLTGLLSNFVSQLSIRNYLYNSTVFEDIMSWVVAMSSSTMRPIRHTATVFCLNIMTFLCEKSKELLNEHAIATKQLEKEEKRSRVNRNRINELNNSLGEIVKQQDTLTTYLNDYFDSVFVHRYRDVEPKIRVDCLQELGVWINTVPSIFFSGSYLRYLGWMLSDINTTVRLTVVKVLRKFFETDSFIGGLRHFSSRFKERILEMSCVDADIGVRVASIRLCNAMRTCGFLENSEILKVLKLILDINPRVQREAVLFLCKVVDESVNEKIDLWGEEDYILKAFSQTSLTTFSVHWIKFSQMCKLLEEVRLSYQSSFDYDTLLRIFQKNGNFITPITQALLNACEIDSIYQSWEDISNFVLFDNYTSTLKDPIDSILSFCKLNDFQESILLQLLSASIQTVCNNNFITPKTVHNKQAAETTNDQNKDKDLLYLNLLPYINSITERNSASPTLLHDSLRLLFSMDLTEMTDPQLSRHFELLINNLKKFFLTNNDLQIIQGCTILFLRLDSIPALKEDLKLLVTDICDQTVTEFLKNFGSFNIQDAVITKDEFVIFEACLTRIEGCTSLKDFSDYPEFDIIYERLVSLLSRVPNSYEDTLKFSAINTLQSLLFWFFLRKDNPADEEKKKDDETKVFNCLINIMNNDSSKILQLQAARTFLETVIMKEGVKASHYNDDNRVSEEHNFLKPQFLDALLKILEGWLYTYAKVGQFPFKRLTQASSPHTQISLDKNPLNRRLLEHVCCDLTSKLLIVVSLSNTITPEFSQQFCELRGHYGPKLSAIVDEFLN F45C2C2A2E0443CC F45C2C2A2E0443CC hsk1-swi6-1 3 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait c-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA 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 1A We confirmed that Swi6 interacts with both Hsk1 and Dfp1 through co-immunoprecipitation experiments him1-swi6-1 3 6 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait c-terminus 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 physical association physical association Fig 1B, S1E hsk1-swi6-2 1 6 enzyme target enzyme target substrate substrate unspecified role unspecified role n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-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 enzyme unspecified role unspecified role P50582:p.Ser314Ile mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain S I Wed May 02 14:55:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser314ile' Tue Jun 30 19:48:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser314Ile' c-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA 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 phosphorylation phosphorylation reaction phosphorylation Fig 1D Phosphorylation of swi6. In vitro kinase assays directly demonstrated that recombinant Swi6 is a substrate for wild-type Hsk1−Dfp1 psc3-him1-1 2 7 unspecified role unspecified role bait bait c-terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role ancillary ancillary 4 unspecified role unspecified role prey prey c-terminus 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 association association Fig 4C We therefore examined whether Dfp1 interacts with Psc3 and whether this interaction requires Swi6. Psc3 coprecipitated with Dfp1, demonstrating that these proteins interact him1-swi6-2 3 4 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain c-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role prey prey physical association physical association Fig S1A