UniProt UniProt http://www.uniprot.org Deformed epidermal autoregulatory factor-1 (DEAF1) interacts with the Ku70 subunit of the DNA-dependent protein kinase complex. Deformed epidermal autoregulatory factor-1 (DEAF1) interacts with the Ku70 subunit of the DNA-dependent protein kinase complex. PloS one 2011 imex curation Jensik PJ., Huggenvik JI., Collard MW. EMSA results (6B), or DNA interactions in 6C/6D not added, except as a comment, as sequence of DNA not given. mcollard@siumed.edu 17-May-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Jensik PJ., Huggenvik JI., Collard MW. EMSA results (6B), or DNA interactions in 6C/6D not added, except as a comment, as sequence of DNA not given. mcollard@siumed.edu Accepted 2018-APR-16 AT 14:25 BST AT 14:25 BST by LIVIAP PloS one 2011 Only protein-protein interactions imex curation imex curation Deformed epidermal autoregulatory factor-1 (DEAF1) interacts with the Ku70 subunit of the DNA-dependent protein kinase complex. Deformed epidermal autoregulatory factor-1 (DEAF1) interacts with the Ku70 subunit of the DNA-dependent protein kinase complex. PloS one 2011 imex curation Jensik PJ., Huggenvik JI., Collard MW. EMSA results (6B), or DNA interactions in 6C/6D not added, except as a comment, as sequence of DNA not given. mcollard@siumed.edu 17-May-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down sequence tag sequence tag identification sequence tag Jensik PJ., Huggenvik JI., Collard MW. EMSA results (6B), or DNA interactions in 6C/6D not added, except as a comment, as sequence of DNA not given. mcollard@siumed.edu Accepted 2018-APR-16 AT 14:25 BST AT 14:25 BST by LIVIAP PloS one 2011 Only protein-protein interactions imex curation imex curation Deformed epidermal autoregulatory factor-1 (DEAF1) interacts with the Ku70 subunit of the DNA-dependent protein kinase complex. Deformed epidermal autoregulatory factor-1 (DEAF1) interacts with the Ku70 subunit of the DNA-dependent protein kinase complex. PloS one 2011 imex curation Jensik PJ., Huggenvik JI., Collard MW. EMSA results (6B), or DNA interactions in 6C/6D not added, except as a comment, as sequence of DNA not given. mcollard@siumed.edu 17-May-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation chlae-cv-1 Monkey African Green kidney fibroblast Grivet cv-1 Monkey African Green kidney fibroblast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Jensik PJ., Huggenvik JI., Collard MW. EMSA results (6B), or DNA interactions in 6C/6D not added, except as a comment, as sequence of DNA not given. Interactions involving Ku80/XRCC5 could not be added as the source of this construct could not be traced. mcollard@siumed.edu Accepted 2018-APR-16 AT 14:25 BST AT 14:25 BST by LIVIAP PloS one 2011 Only protein-protein interactions imex curation imex curation Deformed epidermal autoregulatory factor-1 (DEAF1) interacts with the Ku70 subunit of the DNA-dependent protein kinase complex. Deformed epidermal autoregulatory factor-1 (DEAF1) interacts with the Ku70 subunit of the DNA-dependent protein kinase complex. PloS one 2011 imex curation Jensik PJ., Huggenvik JI., Collard MW. EMSA results (6B), or DNA interactions in 6C/6D not added, except as a comment, as sequence of DNA not given. mcollard@siumed.edu 17-May-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Jensik PJ., Huggenvik JI., Collard MW. EMSA results (6B), or DNA interactions in 6C/6D not added, except as a comment, as sequence of DNA not given. Interactions involving Ku80/XRCC5 could not be added as the source of this construct could not be traced. mcollard@siumed.edu Accepted 2018-APR-16 AT 14:25 BST AT 14:25 BST by LIVIAP PloS one 2011 Only protein-protein interactions imex curation imex curation Deformed epidermal autoregulatory factor-1 (DEAF1) interacts with the Ku70 subunit of the DNA-dependent protein kinase complex. Deformed epidermal autoregulatory factor-1 (DEAF1) interacts with the Ku70 subunit of the DNA-dependent protein kinase complex. PloS one 2011 imex curation Jensik PJ., Huggenvik JI., Collard MW. EMSA results (6B), or DNA interactions in 6C/6D not added, except as a comment, as sequence of DNA not given. mcollard@siumed.edu 17-May-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation chlae-cv-1 Monkey African Green kidney fibroblast Grivet cv-1 Monkey African Green kidney fibroblast anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Jensik PJ., Huggenvik JI., Collard MW. EMSA results (6B), or DNA interactions in 6C/6D not added, except as a comment, as sequence of DNA not given. Interactions involving Ku80/XRCC5 could not be added as the source of this construct could not be traced. mcollard@siumed.edu Accepted 2018-APR-16 AT 14:25 BST AT 14:25 BST by LIVIAP PloS one 2011 Only protein-protein interactions imex curation imex curation Deformed epidermal autoregulatory factor-1 (DEAF1) interacts with the Ku70 subunit of the DNA-dependent protein kinase complex. Deformed epidermal autoregulatory factor-1 (DEAF1) interacts with the Ku70 subunit of the DNA-dependent protein kinase complex. PloS one 2011 imex curation Jensik PJ., Huggenvik JI., Collard MW. EMSA results (6B), or DNA interactions in 6C/6D not added, except as a comment, as sequence of DNA not given. mcollard@siumed.edu 17-May-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Jensik PJ., Huggenvik JI., Collard MW. EMSA results (6B), or DNA interactions in 6C/6D not added, except as a comment, as sequence of DNA not given. mcollard@siumed.edu Accepted 2018-APR-16 AT 14:25 BST AT 14:25 BST by LIVIAP PloS one 2011 Only protein-protein interactions imex curation imex curation Mock publication and experiment for abstract interactions that are not interaction evidences. Mock publication and experiment for abstract interactions that are not interaction evidences. unspecified method xrcc5_human X-ray repair cross-complementing protein 5 XRCC5 G22P2 ATP-dependent DNA helicase II 80 kDa subunit Lupus Ku autoantigen protein p86 86 kDa subunit of Ku antigen Ku86 Ku80 Thyroid-lupus autoantigen CTC box-binding factor 85 kDa subunit Nuclear factor IV DNA repair protein XRCC5 X-ray repair complementing defective repair in Chinese hamster cells 5 (double-strand-break rejoining) ATP-dependent DNA helicase 2 subunit 2 protein protein human Homo sapiens Human MVRSGNKAAVVLCMDVGFTMSNSIPGIESPFEQAKKVITMFVQRQVFAENKDEIALVLFGTDGTDNPLSGGDQYQNITVHRHLMLPDFDLLEDIESKIQPGSQQADFLDALIVSMDVIQHETIGKKFEKRHIEIFTDLSSRFSKSQLDIIIHSLKKCDISLQFFLPFSLGKEDGSGDRGDGPFRLGGHGPSFPLKGITEQQKEGLEIVKMVMISLEGEDGLDEIYSFSESLRKLCVFKKIERHSIHWPCRLTIGSNLSIRIAAYKSILQERVKKTWTVVDAKTLKKEDIQKETVYCLNDDDETEVLKEDIIQGFRYGSDIVPFSKVDEEQMKYKSEGKCFSVLGFCKSSQVQRRFFMGNQVLKVFAARDDEAAAVALSSLIHALDDLDMVAIVRYAYDKRANPQVGVAFPHIKHNYECLVYVQLPFMEDLRQYMFSSLKNSKKYAPTEAQLNAVDALIDSMSLAKKDEKTDTLEDLFPTTKIPNPRFQRLFQCLLHRALHPREPLPPIQQHIWNMLNPPAEVTTKSQIPLSKIKTLFPLIEAKKKDQVTAQEIFQDNHEDGPTAKKLKTEQGGAHFSVSSLAEGSVTSVGSVNPAENFRVLVKQKKASFEEASNQLINHIEQFLDTNETPYFMKSIDCIRAFREEAIKFSEEQRFNNFLKALQEKVEIKQLNHFWEIVVQDGITLITKEEASGSSVTAEEAKKFLAPKDKPSGDTAAVFEEGGDVDDLLDMI DIP protein P13010 original sequence version: 128 DIP protein P13010 original sequence version: 127 DIP protein P13010 original sequence version: 122 DIP protein P13010 original sequence version: 174 DIP protein P13010 original sequence version: 153 DIP protein P13010 original sequence version: 155 DIP protein P13010 original sequence version: 142 2363CA84834E74A3 xrcc6_human X-ray repair cross-complementing protein 6 XRCC6 G22P1 ATP-dependent DNA helicase II 70 kDa subunit Lupus Ku autoantigen protein p70 70 kDa subunit of Ku antigen Thyroid-lupus autoantigen CTC box-binding factor 75 kDa subunit DNA repair protein XRCC6 X-ray repair complementing defective repair in Chinese hamster cells 6 ATP-dependent DNA helicase 2 subunit 1 5'-deoxyribose-5-phosphate lyase Ku70 protein protein human Homo sapiens Human MSGWESYYKTEGDEEAEEEQEENLEASGDYKYSGRDSLIFLVDASKAMFESQSEDELTPFDMSIQCIQSVYISKIISSDRDLLAVVFYGTEKDKNSVNFKNIYVLQELDNPGAKRILELDQFKGQQGQKRFQDMMGHGSDYSLSEVLWVCANLFSDVQFKMSHKRIMLFTNEDNPHGNDSAKASRARTKAGDLRDTGIFLDLMHLKKPGGFDISLFYRDIISIAEDEDLRVHFEESSKLEDLLRKVRAKETRKRALSRLKLKLNKDIVISVGIYNLVQKALKPPPIKLYRETNEPVKTKTRTFNTSTGGLLLPSDTKRSQIYGSRQIILEKEETEELKRFDDPGLMLMGFKPLVLLKKHHYLRPSLFVYPEESLVIGSSTLFSALLIKCLEKEVAALCRYTPRRNIPPYFVALVPQEEELDDQKIQVTPPGFQLVFLPFADDKRKMPFTEKIMATPEQVGKMKAIVEKLRFTYRSDSFENPVLQQHFRNLEALALDLMEPEQAVDLTLPKVEAMNKRLGSLVDEFKELVYPPDYNPEGKVTKRKHDNEGSGSKRPKVEYSEEELKTHISKGTLGKFTVPMLKEACRAYGLKSGLKKQELLEALTKHFQD DIP protein P12956 original sequence version: 184 DIP protein P12956 original sequence version: 138 DIP protein P12956 original sequence version: 185 DIP protein P12956 original sequence version: 164 DIP protein P12956 original sequence version: 166 BBD3CD434526DFCB deaf1_human Deformed epidermal autoregulatory factor 1 homolog Nuclear DEAF-1-related transcriptional regulator Suppressin Zinc finger MYND domain-containing protein 5 DEAF1 SPN ZMYND5 protein protein human Homo sapiens Human MEDSDSAAKQLGLAEAAAVAAAAAVAAAAAAAAGGEAEEPVLSRDEDSEEDADSEAERETPRVTAVAVMAAEPGHMDMGAEALPGPDEAAAAAAFAEVTTVTVANVGAAADNVFTTSVANAASISGHVLSGRTALQIGDSLNTEKATLIVVHTDGSIVETTGLKGPAAPLTPGPQSPPTPLAPGQEKGGTKYNWDPSVYDSELPVRCRNISGTLYKNRLGSGGRGRCIKQGENWYSPTEFEAMAGRASSKDWKRSIRYAGRPLQCLIQDGILNPHAASCTCAACCDDMTLSGPVRLFVPYKRRKKENELPTTPVKKDSPKNITLLPATAATTFTVTPSGQITTSGALTFDRASTVEATAVISESPAQGDVFAGATVQEASVQPPCRASHPEPHYPGYQDSCQIAPFPEAALPTSHPKIVLTSLPALAVPPPTPTKAAPPALVNGLELSEPRSWLYLEEMVNSLLNTAQQLKTLFEQAKHASTYREAATNQAKIHADAERKEQSCVNCGREAMSECTGCHKVNYCSTFCQRKDWKDHQHICGQSAAVTVQADEVHVAESVMEKVTV 3BDFEDBF6AD4BDDE prkdc_human DNA-dependent protein kinase catalytic subunit PRKDC HYRC HYRC1 DNPK1 p460 protein protein human Homo sapiens Human MAGSGAGVRCSLLRLQETLSAADRCGAALAGHQLIRGLGQECVLSSSPAVLALQTSLVFSRDFGLLVFVRKSLNSIEFRECREEILKFLCIFLEKMGQKIAPYSVEIKNTCTSVYTKDRAAKCKIPALDLLIKLLQTFRSSRLMDEFKIGELFSKFYGELALKKKIPDTVLEKVYELLGLLGEVHPSEMINNAENLFRAFLGELKTQMTSAVREPKLPVLAGCLKGLSSLLCNFTKSMEEDPQTSREIFNFVLKAIRPQIDLKRYAVPSAGLRLFALHASQFSTCLLDNYVSLFEVLLKWCAHTNVELKKAALSALESFLKQVSNMVAKNAEMHKNKLQYFMEQFYGIIRNVDSNNKELSIAIRGYGLFAGPCKVINAKDVDFMYVELIQRCKQMFLTQTDTGDDRVYQMPSFLQSVASVLLYLDTVPEVYTPVLEHLVVMQIDSFPQYSPKMQLVCCRAIVKVFLALAAKGPVLRNCISTVVHQGLIRICSKPVVLPKGPESESEDHRASGEVRTGKWKVPTYKDYVDLFRHLLSSDQMMDSILADEAFFSVNSSSESLNHLLYDEFVKSVLKIVEKLDLTLEIQTVGEQENGDEAPGVWMIPTSDPAANLHPAKPKDFSAFINLVEFCREILPEKQAEFFEPWVYSFSYELILQSTRLPLISGFYKLLSITVRNAKKIKYFEGVSPKSLKHSPEDPEKYSCFALFVKFGKEVAVKMKQYKDELLASCLTFLLSLPHNIIELDVRAYVPALQMAFKLGLSYTPLAEVGLNALEEWSIYIDRHVMQPYYKDILPCLDGYLKTSALSDETKNNWEVSALSRAAQKGFNKVVLKHLKKTKNLSSNEAISLEEIRIRVVQMLGSLGGQINKNLLTVTSSDEMMKSYVAWDREKRLSFAVPFREMKPVIFLDVFLPRVTELALTASDRQTKVAACELLHSMVMFMLGKATQMPEGGQGAPPMYQLYKRTFPVLLRLACDVDQVTRQLYEPLVMQLIHWFTNNKKFESQDTVALLEAILDGIVDPVDSTLRDFCGRCIREFLKWSIKQITPQQQEKSPVNTKSLFKRLYSLALHPNAFKRLGASLAFNNIYREFREEESLVEQFVFEALVIYMESLALAHADEKSLGTIQQCCDAIDHLCRIIEKKHVSLNKAKKRRLPRGFPPSASLCLLDLVKWLLAHCGRPQTECRHKSIELFYKFVPLLPGNRSPNLWLKDVLKEEGVSFLINTFEGGGCGQPSGILAQPTLLYLRGPFSLQATLCWLDLLLAALECYNTFIGERTVGALQVLGTEAQSSLLKAVAFFLESIAMHDIIAAEKCFGTGAAGNRTSPQEGERYNYSKCTVVVRIMEFTTTLLNTSPEGWKLLKKDLCNTHLMRVLVQTLCEPASIGFNIGDVQVMAHLPDVCVNLMKALKMSPYKDILETHLREKITAQSIEELCAVNLYGPDAQVDRSRLAAVVSACKQLHRAGLLHNILPSQSTDLHHSVGTELLSLVYKGIAPGDERQCLPSLDLSCKQLASGLLELAFAFGGLCERLVSLLLNPAVLSTASLGSSQGSVIHFSHGEYFYSLFSETINTELLKNLDLAVLELMQSSVDNTKMVSAVLNGMLDQSFRERANQKHQGLKLATTILQHWKKCDSWWAKDSPLETKMAVLALLAKILQIDSSVSFNTSHGSFPEVFTTYISLLADTKLDLHLKGQAVTLLPFFTSLTGGSLEELRRVLEQLIVAHFPMQSREFPPGTPRFNNYVDCMKKFLDALELSQSPMLLELMTEVLCREQQHVMEELFQSSFRRIARRGSCVTQVGLLESVYEMFRKDDPRLSFTRQSFVDRSLLTLLWHCSLDALREFFSTIVVDAIDVLKSRFTKLNESTFDTQITKKMGYYKILDVMYSRLPKDDVHAKESKINQVFHGSCITEGNELTKTLIKLCYDAFTENMAGENQLLERRRLYHCAAYNCAISVICCVFNELKFYQGFLFSEKPEKNLLIFENLIDLKRRYNFPVEVEVPMERKKKYIEIRKEAREAANGDSDGPSYMSSLSYLADSTLSEEMSQFDFSTGVQSYSYSSQDPRPATGRFRRREQRDPTVHDDVLELEMDELNRHECMAPLTALVKHMHRSLGPPQGEEDSVPRDLPSWMKFLHGKLGNPIVPLNIRLFLAKLVINTEEVFRPYAKHWLSPLLQLAASENNGGEGIHYMVVEIVATILSWTGLATPTGVPKDEVLANRLLNFLMKHVFHPKRAVFRHNLEIIKTLVECWKDCLSIPYRLIFEKFSGKDPNSKDNSVGIQLLGIVMANDLPPYDPQCGIQSSEYFQALVNNMSFVRYKEVYAAAAEVLGLILRYVMERKNILEESLCELVAKQLKQHQNTMEDKFIVCLNKVTKSFPPLADRFMNAVFFLLPKFHGVLKTLCLEVVLCRVEGMTELYFQLKSKDFVQVMRHRDDERQKVCLDIIYKMMPKLKPVELRELLNPVVEFVSHPSTTCREQMYNILMWIHDNYRDPESETDNDSQEIFKLAKDVLIQGLIDENPGLQLIIRNFWSHETRLPSNTLDRLLALNSLYSPKIEVHFLSLATNFLLEMTSMSPDYPNPMFEHPLSECEFQEYTIDSDWRFRSTVLTPMFVETQASQGTLQTRTQEGSLSARWPVAGQIRATQQQHDFTLTQTADGRSSFDWLTGSSTDPLVDHTSPSSDSLLFAHKRSERLQRAPLKSVGPDFGKKRLGLPGDEVDNKVKGAAGRTDLLRLRRRFMRDQEKLSLMYARKGVAEQKREKEIKSELKMKQDAQVVLYRSYRHGDLPDIQIKHSSLITPLQAVAQRDPIIAKQLFSSLFSGILKEMDKFKTLSEKNNITQKLLQDFNRFLNTTFSFFPPFVSCIQDISCQHAALLSLDPAAVSAGCLASLQQPVGIRLLEEALLRLLPAELPAKRVRGKARLPPDVLRWVELAKLYRSIGEYDVLRGIFTSEIGTKQITQSALLAEARSDYSEAAKQYDEALNKQDWVDGEPTEAEKDFWELASLDCYNHLAEWKSLEYCSTASIDSENPPDLNKIWSEPFYQETYLPYMIRSKLKLLLQGEADQSLLTFIDKAMHGELQKAILELHYSQELSLLYLLQDDVDRAKYYIQNGIQSFMQNYSSIDVLLHQSRLTKLQSVQALTEIQEFISFISKQGNLSSQVPLKRLLNTWTNRYPDAKMDPMNIWDDIITNRCFFLSKIEEKLTPLPEDNSMNVDQDGDPSDRMEVQEQEEDISSLIRSCKFSMKMKMIDSARKQNNFSLAMKLLKELHKESKTRDDWLVSWVQSYCRLSHCRSRSQGCSEQVLTVLKTVSLLDENNVSSYLSKNILAFRDQNILLGTTYRIIANALSSEPACLAEIEEDKARRILELSGSSSEDSEKVIAGLYQRAFQHLSEAVQAAEEEAQPPSWSCGPAAGVIDAYMTLADFCDQQLRKEEENASVIDSAELQAYPALVVEKMLKALKLNSNEARLKFPRLLQIIERYPEETLSLMTKEISSVPCWQFISWISHMVALLDKDQAVAVQHSVEEITDNYPQAIVYPFIISSESYSFKDTSTGHKNKEFVARIKSKLDQGGVIQDFINALDQLSNPELLFKDWSNDVRAELAKTPVNKKNIEKMYERMYAALGDPKAPGLGAFRRKFIQTFGKEFDKHFGKGGSKLLRMKLSDFNDITNMLLLKMNKDSKPPGNLKECSPWMSDFKVEFLRNELEIPGQYDGRGKPLPEYHVRIAGFDERVTVMASLRRPKRIIIRGHDEREHPFLVKGGEDLRQDQRVEQLFQVMNGILAQDSACSQRALQLRTYSVVPMTSRLGLIEWLENTVTLKDLLLNTMSQEEKAAYLSDPRAPPCEYKDWLTKMSGKHDVGAYMLMYKGANRTETVTSFRKRESKVPADLLKRAFVRMSTSPEAFLALRSHFASSHALICISHWILGIGDRHLNNFMVAMETGGVIGIDFGHAFGSATQFLPVPELMPFRLTRQFINLMLPMKETGLMYSIMVHALRAFRSDPGLLTNTMDVFVKEPSFDWKNFEQKMLKKGGSWIQEINVAEKNWYPRQKICYAKRKLAGANPAVITCDELLLGHEKAPAFRDYVAVARGSKDHNIRAQEPESGLSEETQVKCLMDQATDPNILGRTWEGWEPWM DIP protein P78527 original sequence version: 118 DIP protein P78527-1 original sequence version: 117 DIP protein P78527 original sequence version: 170 DIP protein P78527 original sequence version: 179 DIP protein P78527 original sequence version: 114 DIP protein P78527 original sequence version: 125 AC6E747FEB09F3E5 deaf1-xrcc5-1 2 8 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level human-pc3 Human PC3 Caucasian prostate adenocarcinoma pc3 Human Caucasian prostate adenocarcinoma 9 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level human-pc3 Human PC3 Caucasian prostate adenocarcinoma pc3 Human Caucasian prostate adenocarcinoma 10 unspecified role unspecified role bait bait purified purified 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 ecoli Escherichia coli (strain K12) association association Fig. 1A, Supp Fig. 1 deaf1-xrcc5-2 6 11 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level human-pc3 Human PC3 Caucasian prostate adenocarcinoma pc3 Human Caucasian prostate adenocarcinoma 10 unspecified role unspecified role bait bait purified purified 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 1B To examine the potential interaction with this complex, GST-DEAF1 pull-downs were repeated and Western blot analysis was performed using an antibody against DNA-PKcs. xrcc5-deaf1-1 3 9 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell 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 Fig. 1C DEAF1-FLAG was able to coimmunoprecipitate both HA-Ku70 and Ku80-HA (Fig. 1C), and reversing the epitopes showed that both FLAG-Ku70 and FLAG-Ku80 were able to coimmunoprecipitate DEAF1 (Fig. 1D). deaf1-xrcc6-1 5 9 unspecified role unspecified role bait bait cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level 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 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell physical association physical association Fig. 1D DEAF1-FLAG was able to coimmunoprecipitate both HA-Ku70 and Ku80-HA (Fig. 1C), and reversing the epitopes showed that both FLAG-Ku70 and FLAG-Ku80 were able to coimmunoprecipitate DEAF1 (Fig. 1D). deaf1-xrcc6-2 4 9 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 10 unspecified role unspecified role bait bait purified purified 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 3C deaf1-xrcc6-3 4 10 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 9 unspecified role unspecified role bait bait purified purified 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 2A, 4 DEAF1-FLAG was able to coimmunoprecipitate both HA-Ku70 and Ku80-HA (Fig. 1C), and reversing the epitopes showed that both FLAG-Ku70 and FLAG-Ku80 were able to coimmunoprecipitate DEAF1 (Fig. 1D). xrcc6-deaf1-1 4 10 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 9 unspecified role unspecified role bait bait purified purified 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 50 46 physical association physical association Fig. 2B The interaction domains of Ku70 and DEAF1 were further delineated by using various GST-Ku70 fusion proteins in pull-downs with two in vitro translated DEAF1 peptides: an amino-terminal (N-terminal) deletion of DEAF1 (amino acids 167–565) (Fig. 3A) and an internal peptide (amino acids 155–326) (Fig. 3B). deaf1-dna_pk_human-1 1 53 enzyme enzyme unspecified role unspecified role purified purified human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 10 enzyme target enzyme target substrate substrate unspecified role unspecified role purified purified 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 ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation To investigate the possibility that DEAF1 is a substrate for DNA-PK, in vitro kinase assays were performed using DNA-PK with GST or GST-DEAF1 (Fig. 6A). Fig. 6A DNA-PK phosphorylated GST-DEAF1 both in the absence and presence of dsDNA xrcc6-deaf1-2 4 10 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 9 unspecified role unspecified role bait bait purified purified 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association Fig. 6C, 6D DNA with DEAF1 binding sites can displace or prevent Ku70 from binding to DEAF1 dna-pk_human DNA-PK PRKDC:XRCC5:XRCC6 PRKDC:XRCC5:XRCC6 heterotrimer DNA-PKc:Ku70:Ku80 heterotrimer DNA-dependent protein kinase complex 64 unspecified role unspecified role 11 enzyme enzyme physical association physical association human Homo sapiens Human stable complex stable complex physical interaction evidence used in manual assertion physical interaction evidence used in manual assertion IPI inferred from physical interaction Serine/threonine-protein kinase complex, central to the nonhomologous end joining (NHEJ) DNA-repair pathway, which rejoins double-strand breaks. It is also required for V(D)J recombination, a process that utilizes NHEJ to promote immune system diversity. Directly impacts transcriptional regulation, in part potentially by phosphorylating the C-terminal domain of RNA polymerse II. The XRCC5/6 heterodimer binds ends of dsDNA and both recruits PRKDC to DNA double strand breaks and stimulates its kinase activity. The structure of PRKDC consists of an N-terminal region, a circular cradle unit, and the kinase head domain. The N-terminal L-shaped region is further divided into the arm (residues 1-400) and the bridge (residues 401-890). The bridge connects to the top of the circular cradle and contacts the C-terminal head region. A front aperture is formed by the circular cradle and a bottom aperture is formed by the arm and the bottom part of the circular cradle and both are large enough to allow the passage of dsDNA. The XRCC5 globular domain binds at the arm region of PRKDCs, which is adjacent to the general location of the XRCC6/XRCC5 DNA-binding domains. The long flexible linker between the XRCC5 C-terminal region and the core XRCC6/XRCC5 DNA-binding domains may allow concerted DNA binding and XRCC translocation. Heterotrimer Accepted 2018-APR-09 by LIVIAP ku70_80_human Ku70:Ku80 heterodimer Ku70:Ku80 complex Ku70:Ku86 complex G22P1:G22P2 complex XRCC5:XRCC6 Ku complex 8 unspecified role unspecified role xrcc6 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 9 unspecified role unspecified role xrcc5 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 67 69 direct interaction direct interaction human Homo sapiens Human stable complex stable complex physical interaction evidence used in manual assertion physical interaction evidence used in manual assertion IPI inferred from physical interaction Single-stranded DNA-dependent 3-prime to 5-prime ATP-dependent helicase complex which binds preferentially to fork-like ends of double-stranded DNA in a cell cycle-dependent manner. Also has 5-prime-deoxyribose-5-phosphate lyase activity, nicking DNA 3-prime of an a basic site by a mechanism involving a Schiff base covalent intermediate with the a basic site. However, its main function appears to be the recruitment of several factors with enzymatic activities to DNA double stranded breaks (DSBs). Required for DSB repair, chromosome maintenance, transcription regulation, V(D)J recombination, and activating DNA-PK. Forms a dyad-symmetrical molecule with a preformed ring that encircles duplex DNA. The binding site can cradle two full turns of DNA while encircling only the central 3-4 base pairs. A broken DNA end is able to thread through the channel formed by the complex, whilst unbroken DNA cannot bind in this manner, ensuring exquisitely specific DSB recognition. Heterodimer created with intact-jami-dbscripts