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/ The RIDDLE syndrome protein mediates a ubiquitin-dependent signaling cascade at sites of DNA damage. The RIDDLE syndrome protein mediates a ubiquitin-dependent signaling cascade at sites of DNA damage. Cell (0092-8674) 2009 imex curation Stewart GS., Panier S., Townsend K., Al-Hakim AK., Kolas NK., Miller ES., Nakada S., Ylanko J., Olivarius S., Mendez M., Oldreive C., Wildenhain J., Tagliaferro A., Pelletier L., Taubenheim N., Durandy A., Byrd PJ., Stankovic T., Taylor AM., Durocher D. g.s.stewart@bham.ac.uk 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 5 human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference confocal microscopy confocal microscopy immunostaining immunostaining Stewart GS., Panier S., Townsend K., Al-Hakim AK., Kolas NK., Miller ES., Nakada S., Ylanko J., Olivarius S., Mendez M., Oldreive C., Wildenhain J., Tagliaferro A., Pelletier L., Taubenheim N., Durandy A., Byrd PJ., Stankovic T., Taylor AM., Durocher D. g.s.stewart@bham.ac.uk Accepted 2009-APR-25 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions 5 imex curation The RIDDLE syndrome protein mediates a ubiquitin-dependent signaling cascade at sites of DNA damage. The RIDDLE syndrome protein mediates a ubiquitin-dependent signaling cascade at sites of DNA damage. Cell (0092-8674) 2009 imex curation Stewart GS., Panier S., Townsend K., Al-Hakim AK., Kolas NK., Miller ES., Nakada S., Ylanko J., Olivarius S., Mendez M., Oldreive C., Wildenhain J., Tagliaferro A., Pelletier L., Taubenheim N., Durandy A., Byrd PJ., Stankovic T., Taylor AM., Durocher D. g.s.stewart@bham.ac.uk 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 5 human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 confocal microscopy confocal microscopy immunostaining immunostaining Stewart GS., Panier S., Townsend K., Al-Hakim AK., Kolas NK., Miller ES., Nakada S., Ylanko J., Olivarius S., Mendez M., Oldreive C., Wildenhain J., Tagliaferro A., Pelletier L., Taubenheim N., Durandy A., Byrd PJ., Stankovic T., Taylor AM., Durocher D. Tagged proteins were visualised by tag fluorescence. g.s.stewart@bham.ac.uk Accepted 2009-APR-25 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions 5 imex curation The RIDDLE syndrome protein mediates a ubiquitin-dependent signaling cascade at sites of DNA damage. The RIDDLE syndrome protein mediates a ubiquitin-dependent signaling cascade at sites of DNA damage. Cell (0092-8674) 2009 imex curation Stewart GS., Panier S., Townsend K., Al-Hakim AK., Kolas NK., Miller ES., Nakada S., Ylanko J., Olivarius S., Mendez M., Oldreive C., Wildenhain J., Tagliaferro A., Pelletier L., Taubenheim N., Durandy A., Byrd PJ., Stankovic T., Taylor AM., Durocher D. g.s.stewart@bham.ac.uk 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 5 human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Stewart GS., Panier S., Townsend K., Al-Hakim AK., Kolas NK., Miller ES., Nakada S., Ylanko J., Olivarius S., Mendez M., Oldreive C., Wildenhain J., Tagliaferro A., Pelletier L., Taubenheim N., Durandy A., Byrd PJ., Stankovic T., Taylor AM., Durocher D. g.s.stewart@bham.ac.uk Accepted 2009-APR-25 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions 5 imex curation The RIDDLE syndrome protein mediates a ubiquitin-dependent signaling cascade at sites of DNA damage. The RIDDLE syndrome protein mediates a ubiquitin-dependent signaling cascade at sites of DNA damage. Cell (0092-8674) 2009 imex curation Stewart GS., Panier S., Townsend K., Al-Hakim AK., Kolas NK., Miller ES., Nakada S., Ylanko J., Olivarius S., Mendez M., Oldreive C., Wildenhain J., Tagliaferro A., Pelletier L., Taubenheim N., Durandy A., Byrd PJ., Stankovic T., Taylor AM., Durocher D. g.s.stewart@bham.ac.uk 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 5 human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Stewart GS., Panier S., Townsend K., Al-Hakim AK., Kolas NK., Miller ES., Nakada S., Ylanko J., Olivarius S., Mendez M., Oldreive C., Wildenhain J., Tagliaferro A., Pelletier L., Taubenheim N., Durandy A., Byrd PJ., Stankovic T., Taylor AM., Durocher D. g.s.stewart@bham.ac.uk Accepted 2009-APR-25 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions 5 imex curation rn168_human E3 ubiquitin-protein ligase RNF168 RNF168 RING-type E3 ubiquitin transferase RNF168 RING finger protein 168 protein protein human Homo sapiens Human MALPKDAIPSLSECQCGICMEILVEPVTLPCNHTLCKPCFQSTVEKASLCCPFCRRRVSSWTRYHTRRNSLVNVELWTIIQKHYPRECKLRASGQESEEVADDYQPVRLLSKPGELRREYEEEISKVAAERRASEEEENKASEEYIQRLLAEEEEEEKRQAEKRRRAMEEQLKSDEELARKLSIDINNFCEGSISASPLNSRKSDPVTPKSEKKSKNKQRNTGDIQKYLTPKSQFGSASHSEAVQEVRKDSVSKDIDSSDRKSPTGQDTEIEDMPTLSPQISLGVGEQGADSSIESPMPWLCACGAEWYHEGNVKTRPSNHGKELCVLSHERPKTRVPYSKETAVMPCGRTESGCAPTSGVTQTNGNNTGETENEESCLLISKEISKRKNQESSFEAVKDPCFSAKRRKVSPESSPDQEETEINFTQKLIDLEHLLFERHKQEEQDRLLALQLQKEVDKEQMVPNRQKGSPDEYHLRATSSPPDKVLNGQRKNPKDGNFKRQTHTKHPTPERGSRDKNRQVSLKMQLKQSVNRRKMPNSTRDHCKVSKSAHSLQPSISQKSVFQMFQRCTK DIP protein Q8IYW5 original sequence version: 81 DIP protein Q8IYW5 original sequence version: 99 DIP protein Q8IYW5 original sequence version: 121 51E16DA92BA654C1 h2ax_human Histone H2AX H2AX H2AFX Histone H2A.X protein protein human Homo sapiens Human MSGRGKTGGKARAKAKSRSSRAGLQFPVGRVHRLLRKGHYAERVGAGAPVYLAAVLEYLTAEILELAGNAARDNKKTRIIPRHLQLAIRNDEELNKLLGGVTIAQGGVLPNIQAVLLPKKTSATVGPKAPSGGKKATQASQEY DIP protein P16104 original sequence version: 114 DIP protein P16104 original sequence version: 110 DIP protein P16104 original sequence version: 117 DIP protein P16104 original sequence version: 133 DIP protein P16104 original sequence version: 140 DIP protein P16104 original sequence version: 113 DIP protein P16104 original sequence version: 99 D4683775C2E6C3A9 ube2n_human Ubiquitin-conjugating enzyme E2 N Ubiquitin-protein ligase N Ubiquitin carrier protein N Ubc13 Bendless-like ubiquitin-conjugating enzyme UBE2N BLU UbcH13 E2 ubiquitin-conjugating enzyme N protein protein human Homo sapiens Human MAGLPRRIIKETQRLLAEPVPGIKAEPDESNARYFHVVIAGPQDSPFEGGTFKLELFLPEEYPMAAPKVRFMTKIYHPNVDKLGRICLDILKDKWSPALQIRTVLLSIQALLSAPNPDDPLANDVAEQWKTNEAQAIETARAWTRLYAMNNI DIP protein P61088 original sequence version: 137 DIP protein P61088 original sequence version: 105 DIP protein P61088 original sequence version: 60 DIP protein P61088 original sequence version: 64 DIP protein P61088 original sequence version: 85 FACD84D883D77407 rnf168-ube2n-1 4 5 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association We detected an interaction of either endogenous (Figure 6E) or epitope-tagged RNF168 (Figure S9D) with UBC13 that was dependent on its RING finger domain. Fig. 6E rnf168-ube2n-2 3 7 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey ring finger region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association We detected an interaction of either endogenous (Figure 6E) or epitope-tagged RNF168 (Figure S9D) with UBC13 that was dependent on its RING finger domain. Supp. Fig. 9D A cys19ser mutant may prevent the interaction from occurring but this protein was under-expressed, making comparison difficult. rnf168-h2ax-1 1 6 unspecified role unspecified role neutral component neutral component ser-140 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role neutral component neutral component colocalization colocalization We found that endogenous RNF168 rapidly relocalized to γ-H2AX-positive subnuclear foci, as soon as 5 min after irradiation (Figure 4A). Fig. 4A, Supp. Figs. 6 - 8 Irradiation (10 Gy) rnf168-h2ax-2 2 6 unspecified role unspecified role neutral component neutral component ser-140 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role neutral component neutral component miu2 region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q8IYW5:p.Ala450Gly mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain A G Wed May 02 19:17:41 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala450gly' Tue Jun 30 19:57:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala450Gly' Q8IYW5:p.[Ala179Gly;Ala450Gly] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain A G certain certain sequence position certain certain certain sequence position certain A G Fri Jul 22 11:00:46 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'ala450gly,ala179gly' Wed May 02 15:36:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala179gly,ala450gly' Tue Jun 30 19:10:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ala179Gly;Ala450Gly]' colocalization colocalization HeLa cells transfected with the indicated GFP-RNF168 expression vectors were irradiated with a 5 Gy IR dose. At the indicated time points, cells were fixed and processed for GFP fluorescence and γ-H2AX immunofluorescence. Fig. 4B Irradiation (10 Gy) rnf168-h2ax-3 1 6 unspecified role unspecified role neutral component neutral component ser-140 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q8IYW5:p.Ala450Gly mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain A G Wed May 02 18:46:03 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala450gly' Tue Jun 30 19:42:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala450Gly' Q8IYW5:p.[Ala179Gly;Ala450Gly] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain A G certain certain sequence position certain certain certain sequence position certain A G Fri Jul 22 11:08:23 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'ala450gly,ala179gly' Wed May 02 17:32:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala179gly,ala450gly' Tue Jun 30 16:48:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ala179Gly;Ala450Gly]' colocalization colocalization HeLa cells transfected with the indicated GFP-RNF168 expression vectors were irradiated with a 5 Gy IR dose. At the indicated time points, cells were fixed and processed for GFP fluorescence and γ-H2AX immunofluorescence. Supp. Fig. 7 Irradiation (10 Gy)