MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Controlling the subcellular localization of DNA polymerases iota and eta via interactions with ubiquitin. Controlling the subcellular localization of DNA polymerases iota and eta via interactions with ubiquitin. EMBO J. (0261-4189) 2006 imex curation Plosky BS., Vidal AE., Fernández de Henestrosa AR., McLenigan MP., McDonald JP., Mead S., Woodgate R. Virus - Publications including interactions involving viral proteins Only protein-protein interactions in vitro In vitro pull down pull down predetermined predetermined participant predetermined Plosky BS., Vidal AE., Fernández de Henestrosa AR., McLenigan MP., McDonald JP., Mead S., Woodgate R. Virus - Publications including interactions involving viral proteins EMBO J. (0261-4189) 2006 Only protein-protein interactions imex curation Controlling the subcellular localization of DNA polymerases iota and eta via interactions with ubiquitin. Controlling the subcellular localization of DNA polymerases iota and eta via interactions with ubiquitin. EMBO J. (0261-4189) 2006 imex curation Plosky BS., Vidal AE., Fernández de Henestrosa AR., McLenigan MP., McDonald JP., Mead S., Woodgate R. Virus - Publications including interactions involving viral proteins Only protein-protein interactions 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 Plosky BS., Vidal AE., Fernández de Henestrosa AR., McLenigan MP., McDonald JP., Mead S., Woodgate R. Virus - Publications including interactions involving viral proteins EMBO J. (0261-4189) 2006 Only protein-protein interactions imex curation dpod3_human DNA polymerase delta subunit 3 DNA polymerase delta subunit p66 POLD3 KIAA0039 DNA polymerase delta subunit C DNA polymerase delta subunit p68 protein protein human Homo sapiens Human MADQLYLENIDEFVTDQNKIVTYKWLSYTLGVHVNQAKQMLYDYVERKRKENSGAQLHVTYLVSGSLIQNGHSCHKVAVVREDKLEAVKSKLAVTASIHVYSIQKAMLKDSGPLFNTDYDILKSNLQNCSKFSAIQCAAAVPRAPAESSSSSKKFEQSHLHMSSETQANNELTTNGHGPPASKQVSQQPKGIMGMFASKAAAKTQETNKETKTEAKEVTNASAAGNKAPGKGNMMSNFFGKAAMNKFKVNLDSEQAVKEEKIVEQPTVSVTEPKLATPAGLKKSSKKAEPVKVLQKEKKRGKRVALSDDETKETENMRKKRRRIKLPESDSSEDEVFPDSPGAYEAESPSPPPPPSPPLEPVPKTEPEPPSVKSSSGENKRKRKRVLKSKTYLDGEGCIVTEKVYESESCTDSEEELNMKTSSVHRPPAMTVKKEPREERKGPKKGTAALGKANRQVSITGFFQRK DIP protein Q15054 original sequence version: 87 E9625E0188725F45 ubc_human Polyubiquitin-C UBC protein protein human Homo sapiens Human MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGV B6E7BC06FEE77196 poli_human DNA polymerase iota Eta2 RAD30 homolog B POLI RAD30B protein protein human Homo sapiens Human MEKLGVEPEEEGGGDDDEEDAEAWAMELADVGAAASSQGVHDQVLPTPNASSRVIVHVDLDCFYAQVEMISNPELKDKPLGVQQKYLVVTCNYEARKLGVKKLMNVRDAKEKCPQLVLVNGEDLTRYREMSYKVTELLEEFSPVVERLGFDENFVDLTEMVEKRLQQLQSDELSAVTVSGHVYNNQSINLLDVLHIRLLVGSQIAAEMREAMYNQLGLTGCAGVASNKLLAKLVSGVFKPNQQTVLLPESCQHLIHSLNHIKEIPGIGYKTAKCLEALGINSVRDLQTFSPKILEKELGISVAQRIQKLSFGEDNSPVILSGPPQSFSEEDSFKKCSSEVEAKNKIEELLASLLNRVCQDGRKPHTVRLIIRRYSSEKHYGRESRQCPIPSHVIQKLGTGNYDVMTPMVDILMKLFRNMVNVKMPFHLTLLSVCFCNLKALNTAKKGLIDYYLMPSLSTTSRSGKHSFKMKDTHMEDFPKDKETNRDFLPSGRIESTRTRESPLDTTNFSKEKDINEFPLCSLPEGVDQEVFKQLPVDIQEEILSGKSREKFQGKGSVSCPLHASRGVLSFFSKKQMQDIPINPRDHLSSSKQVSSVSPCEPGTSGFNSSSSSYMSSQKDYSYYLDNRLKDERISQGPKEPQGFHFTNSNPAVSAFHSFPNLQSEQLFSRNHTTDSHKQTVATDSHEGLTENREPDSVDEKITFPSDIDPQVFYELPEAVQKELLAEWKRAGSDFHIGHK C1A5BF0894E91FDF sumo1_human Small ubiquitin-related modifier 1 SUMO1 Sentrin Ubiquitin-like protein SMT3C Ubiquitin-homology domain protein PIC1 Ubiquitin-like protein UBL1 GAP-modifying protein 1 SMT3 homolog 3 SMT3C SMT3H3 UBL1 OK/SW-cl.43 protein protein human Homo sapiens Human MSDQEAKPSTEDLGDKKEGEYIKLKVIGQDSSEIHFKVKMTTHLKKLKESYCQRQGVPMNSLRFLFEGQRIADNHTPKELGMEEEDVIEVYQEQTGGHSTV DIP protein P63165 original sequence version: 36 DIP protein P63165 original sequence version: 40 DIP protein P63165 original sequence version: 124 DIP protein P63165 original sequence version: 125 DIP protein P63165 original sequence version: 83 DIP protein P63165 original sequence version: 85 DIP protein P63165 original sequence version: 81 DIP protein P63165 original sequence version: 84 DIP protein P63165 original sequence version: 103 DIP protein P63165 original sequence version: 100 DIP protein P63165 original sequence version: 95 DIP protein P63165 original sequence version: 98 DIP protein P63165 original sequence version: 101 DIP protein P63165-1 original sequence version: 131 DIP protein P63165 original sequence version: 102 DIP protein P63165 original sequence version: 122 DIP protein P63165 original sequence version: 62 DIP protein P63165 original sequence version: 119 DIP protein P63165 original sequence version: 82 89BE97D2D054FB33 q6fi35_human Proliferating cell nuclear antigen PCNA protein protein human Homo sapiens Human MFEARLVQGSILKKVLEALKDLINEACWDISSSGVNLQSMDSSHVSLVQLTLRSEGFDTYRCDRNLAMGVNLTSMSKILKCAGNEDIITLRAEDNADTLALVFEAPNQEKVSDYEMKLMDLDVEQLGIPEQEYSCVVKMPSGEFACICRDLSHIGDAVVISCAKDGVKFSASGELGNGNIKLSQTSNVDKEEEAVTIEMNEPVQLTFALRYLNFFTKATPLSSTVTLSMSADVPLVVEYKIADMGHLKYYLAPKIEDEEGS This protein is an auxiliary protein of DNA polymerase delta and is involved in the control of eukaryotic DNA replication by increasing the polymerase's processibility during elongation of the leading strand mint 16E18E726D02506D tdg_human G/T mismatch-specific thymine DNA glycosylase TDG Thymine-DNA glycosylase protein protein human Homo sapiens Human MEAENAGSYSLQQAQAFYTFPFQQLMAEAPNMAVVNEQQMPEEVPAPAPAQEPVQEAPKGRKRKPRTTEPKQPVEPKKPVESKKSGKSAKSKEKQEKITDTFKVKRKVDRFNGVSEAELLTKTLPDILTFNLDIVIIGINPGLMAAYKGHHYPGPGNHFWKCLFMSGLSEVQLNHMDDHTLPGKYGIGFTNMVERTTPGSKDLSSKEFREGGRILVQKLQKYQPRIAVFNGKCIYEIFSKEVFGVKVKNLEFGLQPHKIPDTETLCYVMPSSSARCAQFPRAQDKVHYYIKLKDLRDQLKGIERNMDVQEVQYTFDLQLAQEDAKKMAVKEEKYDPGYEAAYGGAYGENPCSSEPCGFSSNGLIESVELRGESAFSGIPNGQWMTQSFTDQIPSFSNHCGTQEQEEESHA DIP protein Q13569 original sequence version: 70 DIP protein Q13569 original sequence version: 99 33752B26EBC789AE polh_human DNA polymerase eta POLH RAD30 RAD30A XPV RAD30 homolog A Xeroderma pigmentosum variant type protein protein protein human Homo sapiens Human MATGQDRVVALVDMDCFFVQVEQRQNPHLRNKPCAVVQYKSWKGGGIIAVSYEARAFGVTRSMWADDAKKLCPDLLLAQVRESRGKANLTKYREASVEVMEIMSRFAVIERASIDEAYVDLTSAVQERLQKLQGQPISADLLPSTYIEGLPQGPTTAEETVQKEGMRKQGLFQWLDSLQIDNLTSPDLQLTVGAVIVEEMRAAIERETGFQCSAGISHNKVLAKLACGLNKPNRQTLVSHGSVPQLFSQMPIRKIRSLGGKLGASVIEILGIEYMGELTQFTESQLQSHFGEKNGSWLYAMCRGIEHDPVKPRQLPKTIGCSKNFPGKTALATREQVQWWLLQLAQELEERLTKDRNDNDRVATQLVVSIRVQGDKRLSSLRRCCALTRYDAHKMSHDAFTVIKNCNTSGIQTEWSPPLTMLFLCATKFSASAPSSSTDITSFLSSDPSSLPKVPVTSSEAKTQGSGPAVTATKKATTSLESFFQKAAERQKVKEASLSSLTAPTQAPMSNSPSKPSLPFQTSQSTGTEPFFKQKSLLLKQKQLNNSSVSSPQQNPWSNCKALPNSLPTEYPGCVPVCEGVSKLEESSKATPAEMDLAHNSQSMHASSASKSVLEVTQKATPNPSLLAAEDQVPCEKCGSLVPVWDMPEHMDYHFALELQKSFLQPHSSNPQVVSAVSHQGKRNPKSPLACTNKRPRPEGMQTLESFFKPLTH 6D1D35A0F56ECE89 poli-ubc 2 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell P0CG48:p.Gly75Ser mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain G S Tue Jul 12 10:21:45 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 19:21:30 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly75ser' Tue Jun 30 17:11:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly75Ser' range MINT-2789478: original sequence: 75-75 : STLHLVLRLRG tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P0CG48:p.Gly76Ser mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain G S Tue Jul 12 10:21:39 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 16:47:41 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly76ser' Tue Jun 30 17:25:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly76Ser' range MINT-2789481: original sequence: 76-76 : TLHLVLRLRG Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F2A F1A EMBO-Journal e used human poliota as a bait to screen a human testis cDNA library in the yeast two-hybrid system. Approximately 25 30% of the clones that were found to interact with poliota encoded the UbA52 or UbB ubiquitin precursor proteins (Figure 1A and B). homomint mint ubc-poli-1 2 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 5 unspecified role unspecified role prey prey Q9UNA4:p.Pro717Arg mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain P R Wed May 02 17:14:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro717arg' Tue Jun 30 16:14:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro717Arg' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association F3A F3A EMBO-Journal Furthermore, by using a series of poliota deletion mutants, we mapped the region of the Ub-interaction to the C-terminal 224 amino acids of poliota (Figure 1C). homomint mint polh-ubc-1 2 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q9Y253:p.His654Ala mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain H A Tue Jul 12 10:06:18 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 15:15:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his654ala' Tue Jun 30 16:31:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His654Ala' range MINT-2789933: original sequence: 654-654 : VWDMPEHMDYHFALELQKSF Stoichiometry: 1.0 physical association physical association F1D F5A EMBO-Journal Like pol iota, pol eta appears to interact with Ub (Figure 1D). Neither the p125 or p66 subunit of poldelta interacts with Ub, but as expected, the p66 subunit interacts with PCNA (Figure 1D; Hughes et al, 1999). The interaction of poliota and poleta with Ub is therefore specific and at least in the two-hybrid system, does not extend to SUMO-1. We believe that SUMO-1 is active under our assay conditions, as we were able to detect an interaction between SUMO-1 and thymine DNA glycosylase (TDG) (Baba et al, 2005; Figure 1D). Interestingly, a single H654A substitution in one of the conserved histidine residue of the C2H2 motif abolishes the interaction between poleta and Ub (Figure 5A). homomint mint polh-pcna 2 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F1D EMBO-Journal Like pol iota, pol eta appears to interact with Ub (Figure 1D). Neither the p125 or p66 subunit of poldelta interacts with Ub, but as expected, the p66 subunit interacts with PCNA (Figure 1D; Hughes et al, 1999). The interaction of poliota and poleta with Ub is therefore specific and at least in the two-hybrid system, does not extend to SUMO-1. We believe that SUMO-1 is active under our assay conditions, as we were able to detect an interaction between SUMO-1 and thymine DNA glycosylase (TDG) (Baba et al, 2005; Figure 1D). homomint mint poli-pcna 2 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F1D EMBO-Journal Like pol iota, pol eta appears to interact with Ub (Figure 1D). Neither the p125 or p66 subunit of poldelta interacts with Ub, but as expected, the p66 subunit interacts with PCNA (Figure 1D; Hughes et al, 1999). The interaction of poliota and poleta with Ub is therefore specific and at least in the two-hybrid system, does not extend to SUMO-1. We believe that SUMO-1 is active under our assay conditions, as we were able to detect an interaction between SUMO-1 and thymine DNA glycosylase (TDG) (Baba et al, 2005; Figure 1D). homomint mint pold3-pcna-1 2 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 3 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F1D EMBO-Journal Like pol iota, pol eta appears to interact with Ub (Figure 1D). Neither the p125 or p66 subunit of poldelta interacts with Ub, but as expected, the p66 subunit interacts with PCNA (Figure 1D; Hughes et al, 1999). The interaction of poliota and poleta with Ub is therefore specific and at least in the two-hybrid system, does not extend to SUMO-1. We believe that SUMO-1 is active under our assay conditions, as we were able to detect an interaction between SUMO-1 and thymine DNA glycosylase (TDG) (Baba et al, 2005; Figure 1D). homomint mint pcna-polh 2 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F3A EMBO-Journal homomint mint tdg-sumo1 2 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F1D EMBO-Journal Like pol iota, pol eta appears to interact with Ub (Figure 1D). Neither the p125 or p66 subunit of poldelta interacts with Ub, but as expected, the p66 subunit interacts with PCNA (Figure 1D; Hughes et al, 1999). The interaction of poliota and poleta with Ub is therefore specific and at least in the two-hybrid system, does not extend to SUMO-1. We believe that SUMO-1 is active under our assay conditions, as we were able to detect an interaction between SUMO-1 and thymine DNA glycosylase (TDG) (Baba et al, 2005; Figure 1D). homomint mint polh-polh 2 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F3A EMBO-Journal homomint mint polh-ubc-2 2 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 17:08:27 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' 587-641 range MINT-2791297: original sequence: 587-641 : SSKATPAEMDLAHNSQSMHASSASKSVLEVTQKATPNPSLLAAEDQVPCEKCGS binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain range MINT-2789909: original sequence: 484-713 : FQKAAERQKVKEASLSSLTAPTQAPMSNSPSKPSLPFQTSQSTGTEPFFKQKSLLLKQKQLNNSSVSSPQQNPWSNCKALPNSLPTEYPGCVPVCEGVSKLEESSKATPAEMDLAHNSQSMHASSASKSVLEVTQKATPNPSLLAAEDQVPCEKCGSLVPVWDMPEHMDYHFALELQKSFLQPHSSNPQVVSAVSHQGKRNPKSPLACTNKRPRPEGMQTLESFFKPLT tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association SF2 EMBO-Journal by employing the yeast two-hybrid assay and various deletion constructs of poleta, we mapped the Ub-binding site to a region spanning poleta residues 587 641 (Supplementary Figure S2). This region contains a conserved C2H2 or so-called \\'Zinc-finger\\' motif (Vaisman et al, 2004). homomint mint poli-ubc-1 2 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association EMBO-Journal Furthermore, by using a series of poliota deletion mutants, we mapped the region of the Ub-interaction to the C-terminal 224 amino acids of poliota (Figure 1C). FD homomint mint polh-pcna-1 1 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified Stoichiometry: 1.0 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified gst tagged 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 Stoichiometry: 1.0 direct interaction direct interaction F6D EMBO-Journal It is not surprising, therefore, that a GST-fusion corresponding to the C-terminal 130 amino acids of poleta (poleta-[581 713]) can pull-down both unmodified PCNA (Figure 6D, left panel) and Ub-PCNA (Figure 6D, right panel). homomint mint poli-pcna-1 1 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified gst tagged 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 Stoichiometry: 1.0 direct interaction direct interaction F6B EMBO-Journal Using an N-terminal GST-fusion of full-length wild-type poliota, or the poliota P692R mutant, we then performed pull-down assays with PCNA or His-Ub-PCNA. Both poliota and poliota P692R are able to bind to unmodified PCNA (Figure 6B, left panel). homomint mint poli-ubc 1 5 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 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q9UNA4:p.Pro717Arg mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain P R Wed May 02 16:41:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro717arg' Tue Jun 30 19:28:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro717Arg' 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified Stoichiometry: 1.0 direct interaction direct interaction F2B F2B EMBO-Journal As seen in Figure 2B, in the absence of any Ub-conjugating proteins, poliota and poleta preferentially interact with both K48- and K63-linked polyUb 4 7 chains. while peptides encompassing P692 were able to pull-down both K48- and K63-linked polyUb chains, a peptide with the P692R substitution was unable to pull-down either form of Ub chain (Figure 3B). homomint mint polh-ubc-3 1 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified his tagged his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q9Y253:p.His654Ala mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain H A Tue Jul 12 10:49:51 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 17:17:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his654ala' Tue Jun 30 19:17:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His654Ala' range MINT-2789829: original sequence: 654-654 : VWDMPEHMDYHFALELQKSF binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain range MINT-2789836: original sequence: 581-713 : VSKLEESSKATPAEMDLAHNSQSMHASSASKSVLEVTQKATPNPSLLAAEDQVPCEKCGSLVPVWDMPEHMDYHFALELQKSFLQPHSSNPQVVSAVSHQGKRNPKSPLACTNKRPRPEGMQTLESFFKPLT Stoichiometry: 1.0 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell P0CG48:p.Gly76Ser mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain G S Tue Jul 12 10:07:02 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 17:43:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly76ser' Tue Jun 30 18:05:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly76Ser' range MINT-2790297: original sequence: 76-76 : TLHLVLRLRG tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P0CG48:p.Gly75Ser mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain G S Tue Jul 12 10:06:54 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 16:45:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly75ser' Tue Jun 30 20:07:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly75Ser' range MINT-2790294: original sequence: 75-75 : STLHLVLRLRG Stoichiometry: 1.0 physical association physical association F5B F2B EMBO-Journal As seen in Figure 2B, in the absence of any Ub-conjugating proteins, poliota and poleta preferentially interact with both K48- and K63-linked polyUb 4 7 chains. we used full-length C-terminal His-tagged poleta or poleta-H654A and assayed the ability of each to bind Ub. As shown in Figure 5B, the wild-type protein clearly binds K48- and K63-linked polyUb chains. In contrast, the H654A poleta mutant protein is unable to bind either form of polyUb. homomint mint ubc-pcna 1 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified Q6FI35:p.Lys164Arg mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K R Tue Jul 12 10:14:44 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 17:50:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys164arg' Tue Jun 30 17:05:38 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys164Arg' range MINT-2790034: original sequence: 164-164 : IGDAVVISCAKDGVKFSASG Stoichiometry: 1.0 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified his tagged his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 direct interaction direct interaction EMBO-Journal We were therefore interested in assaying the ability of both wild-type poliota and poleta and their respective Ub-binding mutants to interact with Ub-PCNA in vitro. To do so, we first generated His-tagged Ub-PCNA in vitro following the protocol recently described for the ubiquitination of S. cerivisiae PCNA by Garg and Burgers (2005) (Supplementary Figure S3A and Supplementary Materials and Methods). As expected from earlier studies (Hoege et al, 2002), the His-Ub is conjugated to PCNA via K164, since a K164R PCNA mutant could not be ubiquitinated under the same assay conditions in vitro (Supplementary Figure S3B). SF3A homomint mint