IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ http://www.ebi.ac.uk/ stanitsa-2006-2 Vaccinia virus uracil DNA glycosylase interacts with the A20 protein to form a heterodimeric processivity factor for the viral DNA polymerase. chlae-bsc40 Chlorocebus aethiops Grivet bsc40 Cerae kidney epithelial cells http://www.lgcpromochem-atcc.com/common/catalog/numSearch/numResults.cfm?atccNum=CRL-2761 these cells are temperature tolerant anti tag coip anti tag coimmunoprecipitation anti tag coip predetermined predetermined participant predetermined Stanitsa ES., Arps L., Traktman P. BioCreative - Critical Assessment of Information Extraction systems in Biology A coupled in vitro transcription-translation system was used in this study. ptrakt@mcw.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation stanitsa-2006-4 Vaccinia virus uracil DNA glycosylase interacts with the A20 protein to form a heterodimeric processivity factor for the viral DNA polymerase. chlae-bsc40 Chlorocebus aethiops Grivet bsc40 Cerae kidney epithelial cells http://www.lgcpromochem-atcc.com/common/catalog/numSearch/numResults.cfm?atccNum=CRL-2761 these cells are temperature tolerant pull down pull down predetermined predetermined participant predetermined Stanitsa ES., Arps L., Traktman P. BioCreative - Critical Assessment of Information Extraction systems in Biology A coupled in vitro transcription-translation system was used in this study. ptrakt@mcw.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation stanitsa-2006-1 Vaccinia virus uracil DNA glycosylase interacts with the A20 protein to form a heterodimeric processivity factor for the viral DNA polymerase. in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip predetermined predetermined participant predetermined Stanitsa ES., Arps L., Traktman P. BioCreative - Critical Assessment of Information Extraction systems in Biology A coupled in vitro transcription-translation system was used in this study. ptrakt@mcw.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation stanitsa-2006-3 Vaccinia virus uracil DNA glycosylase interacts with the A20 protein to form a heterodimeric processivity factor for the viral DNA polymerase. chlae-bsc40 Chlorocebus aethiops Grivet bsc40 Cerae kidney epithelial cells http://www.lgcpromochem-atcc.com/common/catalog/numSearch/numResults.cfm?atccNum=CRL-2761 these cells are temperature tolerant anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Stanitsa ES., Arps L., Traktman P. BioCreative - Critical Assessment of Information Extraction systems in Biology A coupled in vitro transcription-translation system was used in this study. ptrakt@mcw.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation ung_vaccc Uracil-DNA glycosylase UNG D4R protein protein vaccc Vaccinia virus (strain Copenhagen) (VACV) VACV MNSVTVSHAPYTITYHDDWEPVMSQLVEFYNEVASWLLRDETSPIPDKFFIQLKQPLRNKRVCVCGIDPYPKDGTGVPFESPNFTKKSIKEIASSISRLTGVIDYKGYNLNIIDGVIPWNYYLSCKLGETKSHAIYWDKISKLLLQHITKHVSVLYCLGKTDFSNIRAKLESPVTTIVGYHPAARDRQFEKDRSFEIINVLLELDNKVPINWAQGFIY a20_vaccc DNA polymerase processivity factor component A20 A20R protein protein vaccc Vaccinia virus (strain Copenhagen) (VACV) VACV MTSSADLTNLKELLSLYKSLRFSDSAAIEKYNSLVEWGTSTYWKIGVQKVANVETSISDYYDEVKNKPFNIDPGYYIFLPVYFGSVFIYSKGKNMVELGSGNSFQIPDDMRSACNKVLDSDNGIDFLRFVLLNNRWIMEDAISKYQSPVNIFKLASEYGLNIPKYLEIEIEEDTLFDDELYSIIERSFDDKFPKISISYIKLGELRRQVVDFFKFSFMYIESIKVDRIGDNIFIPSVITKSGKKILVKDVDHLIRSKVREHTFVKVKKKNTFSILYDYDGNGTETRGEVIKRIIDTIGRDYYVNGKYFSKVGSAGLKQLTNKLDINECATVDELVDEINKSGTVKRKIKNQSAFDLSRECLGYPEADFITLVNNMRFKIENCKVVNFNIENTNCLNNPSIETIYRNFNQFVSIFNVVTDVKKRLFE dpol_vaccw DNA polymerase POL VACWR065 E9L protein protein vaccw vaccinia virus (stra Vaccinia virus (strain WR) VACV MDVRCINWFESHGENRFLYLKSRCRNGETVFIRFPHYFYYVVTDEIYQSLSPPPFNARPLGKMRTIDIDETISYNLDIKDRKCSVADMWLIEEPKKRSIQNATMDEFLNISWFYISNGISPDGCYSLDEQYLTKINNGCYHCDDPRNCFAKKIPRFDIPRSYLFLDIECHFDKKFPSVFINPISHTSYCYIDLSGKRLLFTLINEEMLTEQEIQEAVDRGCLRIQSLMEMDYERELVLCSEIVLLRIAKQLLELTFDYVVTFNGHNFDLRYITNRLELLTGEKIIFRSPDKKEAVYLCIYERNQSSHKGVGGMANTTFHVNNNNGTIFFDLYSFIQKSEKLDSYKLDSISKNAFSCMGKVLNRGVREMTFIGDDTTDAKGKAAAFAKVLTTGNYVTVDEDIICKVIRKDIWENGFKVVLLCPTLPNDTYKLSFGKDDVDLAQMYKDYNLNIALDMARYCIHDACLCQYLWEYYGVETKTDAGASTYVLPQSMVFEYRASTVIKGPLLKLLLETKTILVRSETKQKFPYEGGKVFAPKQKMFSNNVLIFDYNSLYPNVCIFGNLSPETLVGVVVSTNRLEEEINNQLLLQKYPPPRYITVHCEPRLPNLISEIAIFDRSIEGTIPRLLRTFLAERARYKKMLKQATSSTEKAIYDSMQYTYKIVANSVYGLMGFRNSALYSYASAKSCTSIGRRMILYLESVLNGAELSNGMLRFANPLSNPFYMDDRDINPIVKTSLPIDYRFRFRSVYGDTDSVFTEIDSQDVDKSIEIAKELERLINNRVLFNNFKIEFEAVYKNLIMQSKKKYTTMKYSASSNSKSVPERINKGTSETRRDVSKFHKNMIKTYKTRLSEMLSEGRMNSNQVCIDILRSLETDLRSEFDSRSSPLELFMLSRMHHSNYKSADNPNMYLVTEYNKNNPETIELGERYYFAYICPANVPWTKKLVNIKTYETIIDRSFKLGSDQRIFYEVYFKRLTSEIVNLLDNKVLCISFFERMFGSKPTFYEA dpol_vaccc DNA polymerase POL E9L protein protein vaccc Vaccinia virus (strain Copenhagen) (VACV) VACV MDVRCINWFESHGENRFLYLKSRCRNGETVFIRFPHYFYYVVTDEIYQSLSPPPFNARPLGKMRTIDIDETISYNLDIKDRKCSVADMWLIEEPKKRSIQNATMDEFLNISWFYISNGISPDGCYSLDEQYLTKINNGCYHCDDPRNCFAKKIPRFDIPRSYLFLDIECHFDKKFPSVFINPISHTSYCYIDLSGKRLLFTLINEEMLTEQEIQEAVDRGCLRIQSLMEMDYERELVLCSEIVLLRIAKQLLELTFDYVVTFNGHNFDLRYITNRLELLTGEKIIFRSPDKKEAVHLCIYERNQSSHKGVGGMANTTFHVNNNNGTIFFDLYSFIQKSEKLDSYKLDSISKNAFSCMGKVLNRGVREMTFIGDDTTDAKGKAAAFAKVLTTGNYVTVDEDIICKVIRKDIWENGFKVVLLCPTLPNDTYKLSFGKDDVDLAQMYKDYNLNIALDMARYCIHDACLCQYLWEYYGVETKTDAGASTYVLPQSMVFEYRASTVIKGPLLKLLLETKTILVRSETKQKFPYEGGKVFAPKQKMFSNNVLIFDYNSLYPNVCIFGNLSPETLVGVVVSTNRLEEEINNQLLLQKYPPPRYITVHCEPRLPNLISEIAIFDRSIEGTIPRLLRTFLAERARYKKMLKQATSSTEKAIYDSMQYTYKIVANSVYGLMGFRNSALYSYASAKSCTSIGRRMILYLESVLNGAELSNGMLRFANPLSNPFYMDDRDINPIVKTSLPIDYRFRFRSVYGDTDSVFTEIDSQDVDKSIEIAKELERLINNRVLFNNFKIEFEAVYKNLIMQSKKKYTTMKYSASSNSKSVPERINKGTSETRRDVSKFHKNMIKTYKTRLSEMLSEGRMNSNQVCIDILRSLETDLRSEFDSRSSPLELFMLSRMHHSNYKSADNPNMYLVTEYNKNNPETIELGERYYFAYICPANVPWTKKLVNIKTYETIIDRSFKLGSDQRIFYEVYFKRLTSEIVNLLDNKVLCISFFERMFGSKPTFYEA ung-a20r-1 Interaction between UNG protein and A20 protein 504168 N/A 504172 unspecified role unspecified role bait bait n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 504173 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false Figure 4C ung-a20r-2 Interaction between UNG protein, E9 protein and A20 protein 504168 N/A 504173 unspecified role unspecified role prey prey N/A 504174 unspecified role unspecified role prey prey N/A 504172 unspecified role unspecified role bait bait n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false direct interaction direct interaction false false false Figure 5A to 5C Proteins were identified by silver staining after SDS PAGE. E9 DNA polymerase (P06856) was not coprecipitated when A20 protein (P20995) was not present. The complex was judged to be extremely stable as washes of up to 750mM NaCL did not remove bound protein (A20) from the column. ung-a20r-3 Interaction between UNG protein, E9 protein and A20 protein 504169 N/A 504175 unspecified role unspecified role prey prey N/A 504173 unspecified role unspecified role prey prey N/A 504172 unspecified role unspecified role bait bait n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false association association false false false Figure 5A to 5C Proteins were identified by silver staining after SDS PAGE. ung-a20r-4 Interaction between UNG protein and A20 protein 504169 N/A 504173 unspecified role unspecified role prey prey N/A 504172 unspecified role unspecified role bait bait n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Figure 5A to 5C Proteins were identified by silver staining after SDS PAGE. ung-a20r Interaction between UNG protein and A20 protein 504170 N/A 504173 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false undefined mutation glu185ala arg186ala ser phe asp189ala asp190ala lys191ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false in vitro In vitro N/A 504172 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false P20536:p.Asp68Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false P20536:p.His181Leu mutation mutation certain certain sequence position certain certain certain sequence position certain false in vitro In vitro direct interaction direct interaction false false false A catalytically inactive mutant of Uracil-DNA glycosylase (P20536) was used in this study. The mutant form of the A20 protein (tsA20-ER5) ( P20995) is a temperature sensetive muatant form of the protein Figure 3 a20r-ung Interaction between A20 protein and UNG protein 504171 N/A 504173 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 504172 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Figure 4C