UniProt UniProt http://www.uniprot.org ho-2001-1 SUMO modification of Rad22, the Schizosaccharomyces pombe homologue of the recombination protein Rad52. schpm Schizosaccharomyces pombe (fission yeast) Fission yeast pull down pull down western blot western blot Immuno blot Immuno blot Ho JC., Warr NJ., Shimizu H., Watts FZ. Apoptosis - Interactions involving proteins with a function related to apoptosis f.z.watts@sussex.ac.uk Nucleic Acids Res. (0305-1048) 2001 Only protein-protein interactions imex curation ho-2001-3 SUMO modification of Rad22, the Schizosaccharomyces pombe homologue of the recombination protein Rad52. in vitro In vitro enzymatic study enzymatic study predetermined predetermined participant predetermined Ho JC., Warr NJ., Shimizu H., Watts FZ. Apoptosis - Interactions involving proteins with a function related to apoptosis Sumoylation of proteins monitored by change in molecular weight of the target protein. f.z.watts@sussex.ac.uk Nucleic Acids Res. (0305-1048) 2001 Only protein-protein interactions imex curation ho-2001-2 SUMO modification of Rad22, the Schizosaccharomyces pombe homologue of the recombination protein Rad52. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Ho JC., Warr NJ., Shimizu H., Watts FZ. Apoptosis - Interactions involving proteins with a function related to apoptosis 3A f.z.watts@sussex.ac.uk Nucleic Acids Res. (0305-1048) 2001 Only protein-protein interactions imex curation pmt3_schpo Ubiquitin-like protein pmt3/smt3 pmt3 smt3 ubl2 SPBC365.06 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSESPSANISDADKSAITPTTGDTSQQDVKPSTEHINLKVVGQDNNEVFFKIKKTTEFSKLMKIYCARQGKSMNSLRFLVDGERIRPDQTPAELDMEDGDQIEAVLEQLGGCTHLCL rad22_schpo DNA repair and recombination protein rad22 rad22 SPAC30D11.10 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSFEQKQHVASEDQGHFNTAYSHEEFNFLQSSLTRKLGPEYVSRRSGPGGFSVSYIESWKAIELANEIFGFNGWSSSIRSINVDFMDENKENGRISLGLSVIVRVTIKDGAYHEDIGYGSIDNCRGKASAFEKCKKEGTTDALKRALRNFGNSLGNCMYDKYYLREVGKMKPPTYHFDSGDLFRKTDPAARESFIKKQKTLNSTRTVNNQPLVNKGEQLAPRRAAELNDEQTREIEMYADEELDNIFVEDDIIAHLAVAEDTAHPAANNHHSEKAGTQINNKDKGSHNSAKPVQRSHTYPVAVPQNTSDSVGNAVTDTSPKTLFDPLKPNTGTPSPKFISARAAAAAEGVVSAPFTNNFNPRLDSPSIRKTSIIDHSKSLPVQRASVLPIIKQSSQTSPVSNNSMIRDSESIINERKENIGLIGVKRSLHDSTTSHNKSDLMRTNSDPQSAMRSRENYDATVDKKAKKG rad51_schpo DNA repair protein rhp51 rhp51 rad51 SPAC644.14c RAD51 homolog protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MADTEVEMQVSAADTNNNENGQAQSNYEYDVNVQDEEDEAAAGPMPLQMLEGNGITASDIKKIHEAGYYTVESIAYTPKRQLLLIKGISEAKADKLLGEASKLVPMGFTTATEYHIRRSELITITTGSKQLDTLLQGGVETGSITELFGEFRTGKSQICHTLAVTCQLPIDMGGGEGKCLYIDTEGTFRPVRLLAVADRYGLNGEEVLDNVAYARAYNADHQLELLQQAANMMSESRFSLLVVDSCTALYRTDFSGRGELSARQMHLARFMRTLQRLADEFGIAVVITNQVVAQVDGISFNPDPKKPIGGNILAHSSTTRLSLRKGRGEQRICKIYDSPCLPESEAIFAINSDGVGDPKEIIAPV 9F26EB9FA4F3C2BA rfa1_schpo Replication factor A protein 1 ssb1 rad11 rpa1 SPBC660.13c Single-stranded DNA-binding protein p68 subunit protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MAERLSVGALRIINTSDASSFPPNPILQVLTVKELNSNPTSGAPKRYRVVLSDSINYAQSMLSTQLNHLVAENKLQKGAFVQLTQFTVNVMKERKILIVLGLNVLTELGVMDKIGNPAGLETVDALRQQQNEQNNASAPRTGISTSTNSFYGNNAAATAPAPPPMMKKPAAPNSLSTIIYPIEGLSPYQNKWTIRARVTNKSEVKHWHNQRGEGKLFSVNLLDESGEIRATGFNDQVDAFYDILQEGSVYYISRCRVNIAKKQYTNVQNEYELMFERDTEIRKAEDQTAVPVAKFSFVSLQEVGDVAKDAVIDVIGVLQNVGPVQQITSRATSRGFDKRDITIVDQTGYEMRVTLWGKTAIEFSVSEESILAFKGVKVNDFQGRSLSMLTSSTMSVDPDIQESHLLKGWYDGQGRGQEFAKHSVISSTLSTTGRSAERKNIAEVQAEHLGMSETPDYFSLKGTIVYIRKKNVSYPACPAADCNKKVFDQGGSWRCEKCNKEYDAPQYRYIITIAVGDHTGQLWLNVFDDVGKLIMHKTADELNDLQENDENAFMNCMAEACYMPYIFQCRAKQDNFKGEMRVRYTVMSINQMDWKEESKRLINFIESAQ 5F2E77A0E12022EA ubc9_schpo SUMO-conjugating enzyme ubc9 hus5 ubc9 SPAC30D11.13 Ubiquitin-conjugating enzyme E2-18 kDa Ubiquitin carrier protein hus5 Ubiquitin carrier protein 9 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSSLCKTRLQEERKQWRRDHPFGFYAKPCKSSDGGLDLMNWKVGIPGKPKTSWEGGLYKLTMAFPEEYPTRPPKCRFTPPLFHPNVYPSGTVCLSILNEEEGWKPAITIKQILLGIQDLLDDPNIASPAQTEAYTMFKKDKVEYEKRVRAQARENAP 4BF3A6389203D27F uba2_schpo Ubiquitin-activating enzyme E1-like uba2 SPBC16H5.03c Pmt3-activating enzyme subunit 2 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MPTLMQLSNDMKPLTFVEALRNFKSAKVLLVGAGGIGCELLKNLLMSGVKEVHIIDLDTIDLSNLNRQFLFRKKHVKQPKAIVAAKTASSFNPNVKLEAYHANIKEDRFNVAWFRQFDLVFNALDNLDARRHVNKQCLLASVPLIESGTTGFLGQVQVIIHGKTECYDCNPKEPPKTYPVCTIRSTPSQPIHCVVWAKSYFFPQLFSNDQESDGIIDNVSANEMERREIAELARETTELNELRSSIGQSDNGFEKIFTKMFTKDIVRLREVPDAWTYRSPPKELSYSELLENAEKATSPWLNEQNVWNVAESFAVLRDSIRRLALRSKSSKDDLSFDKDDKDTLDFVAAAANLRAHVFGIQQLSEFDIKQMAGNIIPAIATTNAVIAGLCITQAIKVLQGDLNDLKNIYLAKRPTRVLHCEKTCKPNPYCPTCSFVLLQLGVNDKNMTLRVLVDDILKSRLHYSEEVSVLNDKLIYDPDFDDNLDKTFDDLGINPAKNTILTVLGDSAVEKDDDGEEATRVPLLIEVTFIDSNSTEGLPYQILSNATSIPLKQQPPSNSPEDSQVLTDEINEVNDFSSSERIVINLDEYDIIVDSKTSSHNKQLKRRPSNDTLTQEAKSKKQAKIHTM AAC9247BB8D2F634 rad31_schpo DNA damage tolerance protein rad31 rad31 SPAC4C5.04 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MGNHNINAEEIALYDRQIRLWGFNAQQALKQSRVLLITASPLANEIAKNLVLSGIGKLCVLDSMTVYEKDVEEQFFIEASDIGQLRANVFKKKLHELNPLVEIDTDTSLISEIDEGKISKFSMVIATQLDYEEFCRINELTRICNASFYATSCFGLYGFAFCDLINHNFAIDRVVDNTKVEEDMFIVQKPMKEAFQSILGETLKPRLAKKIPTLYPAMLSLLKSKKSDPDSIRQVCIEQKLNEKTVLNGEFLSKFSSNISFQWTPVMSVVGGVVSQDALNSISKKQFPIDNFWIFDAESGLAPIYRL pml_human Protein PML PML MYL RNF71 TRIM19 Tripartite motif-containing protein 19 RING finger protein 71 Promyelocytic leukemia protein PP8675 E3 SUMO-protein ligase PML RING-type E3 SUMO transferase PML protein protein human Homo sapiens Human MEPAPARSPRPQQDPARPQEPTMPPPETPSEGRQPSPSPSPTERAPASEEEFQFLRCQQCQAEAKCPKLLPCLHTLCSGCLEASGMQCPICQAPWPLGADTPALDNVFFESLQRRLSVYRQIVDAQAVCTRCKESADFWCFECEQLLCAKCFEAHQWFLKHEARPLAELRNQSVREFLDGTRKTNNIFCSNPNHRTPTLTSIYCRGCSKPLCCSCALLDSSHSELKCDISAEIQQRQEELDAMTQALQEQDSAFGAVHAQMHAAVGQLGRARAETEELIRERVRQVVAHVRAQERELLEAVDARYQRDYEEMASRLGRLDAVLQRIRTGSALVQRMKCYASDQEVLDMHGFLRQALCRLRQEEPQSLQAAVRTDGFDEFKVRLQDLSSCITQGKDAAVSKKASPEAASTPRDPIDVDLPEEAERVKAQVQALGLAEAQPMAVVQSVPGAHPVPVYAFSIKGPSYGEDVSNTTTAQKRKCSQTQCPRKVIKMESEEGKEARLARSSPEQPRPSTSKAVSPPHLDGPPSPRSPVIGSEVFLPNSNHVASGAGEAEERVVVISSSEDSDAENSSSRELDDSSSESSDLQLEGPSTLRVLDENLADPQAEDRPLVFFDLKIDNETQKISQLAAVNRESKFRVVIQPEAFFSIYSKAVSLEVGLQHFLSFLSSMRRPILACYKLWGPGLPNFFRALEDINRLWEFQEAISGFLAALPLIRERVPGASSFKLKNLAQTYLARNMSERSAMAAVLAMRDLCRLLEVSPGPQLAQHVYPFSSLQCFASLQPLVQAAVLPRAEARLLALHNVSFMELLSAHRRDRQGGLKKYSRYLSLQTTTLPPAQPAFNLQALGTYFEGLLEGPALARAEGVSTPLAGRGLAERASQQS rad22-pmt3 Interaction detected by pulldown 493908 N/A 493911 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag monoclonal western monoclonal antibody western blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 493912 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 monoclonal western monoclonal antibody western blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag monoclonal western monoclonal antibody western blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false 2A Both the bait and the prey were over-expressed. rad22-rhp51 Interaction detected by pulldown 493908 N/A 493912 unspecified role unspecified role bait bait region MYC-HIS tagged protein rad22 his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag monoclonal western monoclonal antibody western blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region MYC-HIS tagged protein rad22 myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag monoclonal western monoclonal antibody western blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 493913 unspecified role unspecified role prey prey region HA tagged protein rhp51 ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag monoclonal western monoclonal antibody western blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false 2B Both the bait and the prey were over-expressed. rad22-rpa70-1 Interaction detected by pulldown 493908 N/A 493914 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag monoclonal western monoclonal antibody western blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 493912 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 monoclonal western monoclonal antibody western blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag monoclonal western monoclonal antibody western blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false 2C Both the bait and the prey were over-expressed. rad22-pmt3-1 Interaction detected by pulldown 493908 N/A 493912 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag monoclonal western monoclonal antibody western blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag monoclonal western monoclonal antibody western blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 493911 unspecified role unspecified role prey prey covalent binding covalent binding false false false 3C RAD22 and PMT3 were expressed at wild type levels. rad22-pmt3-3 Interaction detected by pulldown 493908 N/A 493912 unspecified role unspecified role bait bait n terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag monoclonal western monoclonal antibody western blot n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag monoclonal western monoclonal antibody western blot n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 493911 unspecified role unspecified role prey prey n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag monoclonal western monoclonal antibody western blot n-term range n-terminal range n-term range n-term range n-terminal range n-term range false covalent binding covalent binding false false false 2A The experiment was performed in presence of guanidine-HCl providing denaturing conditions, showing that the interaction is in fact a covalent binding. rad22-pmt3-2 Sumoylation of rad22 by hus5 493909 N/A 493915 enzyme enzyme unspecified role unspecified role ecoli Escherichia coli (strain K12) N/A 493912 enzyme target enzyme target substrate substrate unspecified role unspecified role region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 493911 unspecified role unspecified role ancillary ancillary region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag western blot western blot Immuno blot Immuno blot n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) N/A 493916 unspecified role unspecified role ancillary ancillary 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 false ecoli Escherichia coli (strain K12) N/A 493917 unspecified role unspecified role ancillary ancillary 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 false ecoli Escherichia coli (strain K12) sumoylation sumoylation reaction sumoylation false false false 4C The covalent attachment of pmt3 on rad22 requires the presence of rad31, fub2 and hus5. Sumoylation of Rad22 rhp51-pmt3 Sumoylation of rhp51 by hus5 493909 N/A 493911 unspecified role unspecified role ancillary ancillary region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag western blot western blot Immuno blot Immuno blot n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) N/A 493915 enzyme enzyme unspecified role unspecified role ecoli Escherichia coli (strain K12) N/A 493916 unspecified role unspecified role ancillary ancillary 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 false ecoli Escherichia coli (strain K12) N/A 493913 enzyme target enzyme target substrate substrate unspecified role unspecified role region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 493917 unspecified role unspecified role ancillary ancillary 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 false ecoli Escherichia coli (strain K12) sumoylation sumoylation reaction sumoylation false false false 4D The covalent attachment of pmt3 on rad22 requires the presence of rad31, fub2 and hus5. Sumoylation of Rad22 pml-pmt3 Sumoylation of human PML by hus5 493909 N/A 493911 unspecified role unspecified role ancillary ancillary n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag western blot western blot Immuno blot Immuno blot n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 493915 enzyme enzyme unspecified role unspecified role ecoli Escherichia coli (strain K12) N/A 493918 enzyme target enzyme target substrate substrate unspecified role unspecified role region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 493916 unspecified role unspecified role ancillary ancillary 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 false ecoli Escherichia coli (strain K12) N/A 493917 unspecified role unspecified role ancillary ancillary 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 false ecoli Escherichia coli (strain K12) sumoylation sumoylation reaction sumoylation false false false 4B The covalent attachment of pmt3 on PML requires the presence of rad31, fub2 and hus5. Sumoylation of Rad22 Human PML is a well known target for sumoylation and was used to test the S. pombe sumoylation system. rad22-hus5 Interaction detected by anti-tag coIP 493910 N/A 493915 unspecified role unspecified role bait bait n-terminus 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 false ecoli Escherichia coli (strain K12) N/A 493912 unspecified role unspecified role prey prey 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 false ecoli Escherichia coli (strain K12) physical association physical association false false false 3A The relative amount of rad22 which purifies with hus5 is low, suggesting that the interaction between rad22 and hus5 is transiant in nature.