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/ Proliferating cell nuclear antigen (PCNA) may function as a double homotrimer complex in the mammalian cell. Proliferating cell nuclear antigen (PCNA) may function as a double homotrimer complex in the mammalian cell. J. Biol. Chem. (0021-9258) 2005 imex curation Naryzhny SN., Zhao H., Lee H. Proteins crosslinked wuth 1.5% formaldehyde prior to separation by gel filtration. hlee@hrsrh.on.ca 12-MAR-2007: Contacted by JYOTI. Only protein-protein interactions ecobb Escherichia coli (strain B / BL21) crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Naryzhny SN., Zhao H., Lee H. Proteins crosslinked wuth 1.5% formaldehyde prior to separation by gel filtration. hlee@hrsrh.on.ca Accepted 2007-APR-1 by JYOTI. 12-MAR-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 Only protein-protein interactions imex curation Proliferating cell nuclear antigen (PCNA) may function as a double homotrimer complex in the mammalian cell. Proliferating cell nuclear antigen (PCNA) may function as a double homotrimer complex in the mammalian cell. J. Biol. Chem. (0021-9258) 2005 imex curation Naryzhny SN., Zhao H., Lee H. Proteins crosslinked wuth 1.5% formaldehyde prior to separation by gel filtration. hlee@hrsrh.on.ca 12-MAR-2007: Contacted by JYOTI. Only protein-protein interactions crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Naryzhny SN., Zhao H., Lee H. Proteins crosslinked wuth 1.5% formaldehyde prior to separation by gel filtration. hlee@hrsrh.on.ca Accepted 2007-APR-1 by JYOTI. 12-MAR-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 Only protein-protein interactions imex curation Proliferating cell nuclear antigen (PCNA) may function as a double homotrimer complex in the mammalian cell. Proliferating cell nuclear antigen (PCNA) may function as a double homotrimer complex in the mammalian cell. J. Biol. Chem. (0021-9258) 2005 imex curation Naryzhny SN., Zhao H., Lee H. Proteins crosslinked wuth 1.5% formaldehyde prior to separation by gel filtration. hlee@hrsrh.on.ca 12-MAR-2007: Contacted by JYOTI. Only protein-protein interactions crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Naryzhny SN., Zhao H., Lee H. Proteins crosslinked wuth 1.5% formaldehyde prior to separation by gel filtration. hlee@hrsrh.on.ca Accepted 2007-APR-1 by JYOTI. 12-MAR-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 Only protein-protein interactions imex curation dpod1_human DNA polymerase delta catalytic subunit 3'-5' exodeoxyribonuclease DNA polymerase subunit delta p125 POLD1 POLD protein protein human Homo sapiens Human MDGKRRPGPGPGVPPKRARGGLWDDDDAPRPSQFEEDLALMEEMEAEHRLQEQEEEELQSVLEGVADGQVPPSAIDPRWLRPTPPALDPQTEPLIFQQLEIDHYVGPAQPVPGGPPPSRGSVPVLRAFGVTDEGFSVCCHIHGFAPYFYTPAPPGFGPEHMGDLQRELNLAISRDSRGGRELTGPAVLAVELCSRESMFGYHGHGPSPFLRITVALPRLVAPARRLLEQGIRVAGLGTPSFAPYEANVDFEIRFMVDTDIVGCNWLELPAGKYALRLKEKATQCQLEADVLWSDVVSHPPEGPWQRIAPLRVLSFDIECAGRKGIFPEPERDPVIQICSLGLRWGEPEPFLRLALTLRPCAPILGAKVQSYEKEEDLLQAWSTFIRIMDPDVITGYNIQNFDLPYLISRAQTLKVQTFPFLGRVAGLCSNIRDSSFQSKQTGRRDTKVVSMVGRVQMDMLQVLLREYKLRSYTLNAVSFHFLGEQKEDVQHSIITDLQNGNDQTRRRLAVYCLKDAYLPLRLLERLMVLVNAVEMARVTGVPLSYLLSRGQQVKVVSQLLRQAMHEGLLMPVVKSEGGEDYTGATVIEPLKGYYDVPIATLDFSSLYPSIMMAHNLCYTTLLRPGTAQKLGLTEDQFIRTPTGDEFVKTSVRKGLLPQILENLLSARKRAKAELAKETDPLRRQVLDGRQLALKVSANSVYGFTGAQVGKLPCLEISQSVTGFGRQMIEKTKQLVESKYTVENGYSTSAKVVYGDTDSVMCRFGVSSVAEAMALGREAADWVSGHFPSPIRLEFEKVYFPYLLISKKRYAGLLFSSRPDAHDRMDCKGLEAVRRDNCPLVANLVTASLRRLLIDRDPEGAVAHAQDVISDLLCNRIDISQLVITKELTRAASDYAGKQAHVELAERMRKRDPGSAPSLGDRVPYVIISAAKGVAAYMKSEDPLFVLEHSLPIDTQYYLEQQLAKPLLRIFEPILGEGRAEAVLLRGDHTRCKTVLTGKVGGLLAFAKRRNCCIGCRTVLSHQGAVCEFCQPRESELYQKEVSHLNALEERFSRLWTQCQRCQGSLHEDVICTSRDCPIFYMRKKVRKDLEDQEQLLRRFGPPGPEAW 9D04D34AB4AEE810 caf1a_human Chromatin assembly factor 1 subunit A CHAF1A CAF CAF1P150 Chromatin assembly factor I p150 subunit protein protein human Homo sapiens Human MLEELECGAPGARGAATAMDCKDRPAFPVKKLIQARLPFKRLNLVPKGKADDMSDDQGTSVQSKSPDLEASLDTLENNCHVGSDIDFRPKLVNGKGPLDNFLRNRIETSIGQSTVIIDLTEDSNEQPDSLVDHNKLNSEASPSREAINGQREDTGDQQGLLKAIQNDKLAFPGETLSDIPCKTEEEGVGCGGAGRRGDSQECSPRSCPELTSGPRMCPRKEQDSWSEAGGILFKGKVPMVVLQDILAVRPPQIKSLPATPQGKNMTPESEVLESFPEEDSVLSHSSLSSPSSTSSPEGPPAPPKQHSSTSPFPTSTPLRRITKKFVKGSTEKNKLRLQRDQERLGKQLKLRAEREEKEKLKEEAKRAKEEAKKKKEEEKELKEKERREKREKDEKEKAEKQRLKEERRKERQEALEAKLEEKRKKEEEKRLREEEKRIKAEKAEITRFFQKPKTPQAPKTLAGSCGKFAPFEIKEHMVLAPRRRTAFHPDLCSQLDQLLQQQSGEFSFLKDLKGRQPLRSGPTHVSTRNADIFNSDVVIVERGKGDGVPERRKFGRMKLLQFCENHRPAYWGTWNKKTALIRARDPWAQDTKLLDYEVDSDEEWEEEEPGESLSHSEGDDDDDMGEDEDEDDGFFVPHGYLSEDEGVTEECADPENHKVRQKLKAKEWDEFLAKGKRFRVLQPVKIGCVWAADRDCAGDDLKVLQQFAACFLETLPAQEEQTPKASKRERRDEQILAQLLPLLHGNVNGSKVIIREFQEHCRRGLLSNHTGSPRSPSTTYLHTPTPSEDAAIPSKSRLKRLISENSVYEKRPDFRMCWYVHPQVLQSFQQEHLPVPCQWSYVTSVPSAPKEDSGSVPSTGPSQGTPISLKRKSAGSMCITQFMKKRRHDGQIGAEDMDGFQADTEEEEEEEGDCMIVDVPDAAEVQAPCGAASGAGGGVGVDTGKATLTASPLGAS DIP protein Q13111 original sequence version: 98 DIP protein Q13111 original sequence version: 147 DIP protein Q13111 original sequence version: 122 90617F8FE8DB7FD0 pcna_human Proliferating cell nuclear antigen PCNA Cyclin protein protein human Homo sapiens Human MFEARLVQGSILKKVLEALKDLINEACWDISSSGVNLQSMDSSHVSLVQLTLRSEGFDTYRCDRNLAMGVNLTSMSKILKCAGNEDIITLRAEDNADTLALVFEAPNQEKVSDYEMKLMDLDVEQLGIPEQEYSCVVKMPSGEFARICRDLSHIGDAVVISCAKDGVKFSASGELGNGNIKLSQTSNVDKEEEAVTIEMNEPVQLTFALRYLNFFTKATPLSSTVTLSMSADVPLVVEYKIADMGHLKYYLAPKIEDEEGS DIP protein P12004 original sequence version: 200 DIP protein P12004 original sequence version: 128 DIP protein P12004 original sequence version: 139 DIP protein P12004 original sequence version: 185 DIP protein P12004 original sequence version: 102 DIP protein P12004 original sequence version: 105 DIP protein P12004 original sequence version: 158 E6F08E7EDBC48B00 pcna_crigr Proliferating cell nuclear antigen PCNA protein protein crigr Cricetulus griseus (Chinese hamster) Cricetulus barabensis griseus Chinese hamster MFEARLVQGSILKKVLEALKDLINEACWDISSGGVNLQSMDSSHVSLVQLTLRSEGFDTYRCDRNLAMGVNLTSMSKILKCAGNEDIITLRAEDNADTLALVFEAPNQEKVSDYEMKLMDLDVEQLGIPEQEYSCVVKMPSGEFARICRDLSHIGDAVVISCAKDGVKFSASGELGNGNIKLSQTSNVDKEEEAVTIEMNEPVQLTFALRYLNFFTKATPLSPTVTLSMSTDVPLVVEYKISDMGHLKYYLAPKIEDEEGS A521F78743310CF5 pcna-1 2 7 unspecified role unspecified role neutral component neutral component direct interaction direct interaction However, when cells were treated with 1.5% formaldehyde for 30 min to cross-link proteins prior to SDS-PAGE, PCNA was detected at the positions of 33, ~100, and ~200 kDa (Fig. 1A, lane 2). 1 pcna-2 2 7 unspecified role unspecified role neutral component neutral component direct interaction direct interaction However, when cells were treated with 1.5% formaldehyde for 30 min to cross-link proteins prior to SDS-PAGE, PCNA was detected at the positions of 33, ~100, and ~200 kDa (Fig. 1A, lane 2). 1 pcna-3 1 6 unspecified role unspecified role neutral component neutral component direct interaction direct interaction To further confirm that the ~200-kDa PCNA complex does not contain any other proteins, we repeated the cross-linking experiment using purified PCNA as substrate. 1D, 1E pcna-4 1 6 unspecified role unspecified role neutral component neutral component direct interaction direct interaction To further confirm that the ~200-kDa PCNA complex does not contain any other proteins, we repeated the cross-linking experiment using purified PCNA as substrate. 1D, 1E pcna-5 2 6 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 direct interaction direct interaction Consistent with this previous data, the GFP-human PCNA fusion protein produced in CHO cells behaved similarly to endogenous PCNA following crosslinking (Fig. 2, A and B). 2A, 2B pcna-6 2 6 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 P12004:p.[Arg5Ala;Lys110Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain R A Fri Jul 22 11:05:05 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'lys110ala,arg5ala' Wed May 02 20:03:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg5ala,lys110ala' Tue Jun 30 15:54:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg5Ala;Lys110Ala]' direct interaction direct interaction Further mutation analysis identified Arg-5 and Lys-110 as two critical amino acid residues for stable double trimer formation by formaldehyde-mediated cross-linking (Fig. 3A, lanes 8 and 10). 2A, 2B, 3 Synthetic peptides encompassing Arg-5 (1MFEARLVQGSIL12) and Lys-110 (104EAPNQEKV111) inhibited formation of the double trimer but not the trimer. pcna-pold1-1 3 6 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey association association To determine whether PCNA can physically bind to pol {delta} and CAF-1 simultaneously, extracts prepared from cells transfected by wild-type PCNA were subjected to DNase I digestion and immunoprecipitated with PC10. 1D, 1E pcna-pold1-2 3 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait P12004:p.[Arg5Ala;Lys110Ala] mutation mutation certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain R A Fri Jul 22 11:09:19 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'lys110ala,arg5ala' Wed May 02 19:55:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg5ala,lys110ala' Tue Jun 30 20:10:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg5Ala;Lys110Ala]' physical association physical association In contrast, the CAF-1 complex immunoprecipitated from cells transfected by the R5A/K110A mutant construct did not contain pol {delta} (Fig. 4E, lane 8), 4E