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/ Reconstitution of recombinant human replication factor C (RFC) and identification of an RFC subcomplex possessing DNA-dependent ATPase activity. Reconstitution of recombinant human replication factor C (RFC) and identification of an RFC subcomplex possessing DNA-dependent ATPase activity. J. Biol. Chem. (0021-9258) 1998 imex curation Ellison V., Stillman B. Proteins identified by silver staining following SDS-PAGE stillman@cshl.org 08-DEC-2006: Contacted by JYOTI. Only protein-protein interactions spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection solution sedimentati cosedimentation in solution solution sedimentati weight by staining molecular weight estimation by staining weight by staining Ellison V., Stillman B. Proteins identified by silver staining following SDS-PAGE stillman@cshl.org Accepted 2006-NOV-27 by JYOTI. 08-DEC-2006: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 1998 Only protein-protein interactions imex curation Reconstitution of recombinant human replication factor C (RFC) and identification of an RFC subcomplex possessing DNA-dependent ATPase activity. Reconstitution of recombinant human replication factor C (RFC) and identification of an RFC subcomplex possessing DNA-dependent ATPase activity. J. Biol. Chem. (0021-9258) 1998 imex curation Ellison V., Stillman B. Proteins identified by silver staining following SDS-PAGE stillman@cshl.org 08-DEC-2006: Contacted by JYOTI. Only protein-protein interactions spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection pull down pull down weight by staining molecular weight estimation by staining weight by staining Ellison V., Stillman B. Proteins identified by silver staining following SDS-PAGE stillman@cshl.org Accepted 2006-NOV-27 by JYOTI. 08-DEC-2006: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 1998 Only protein-protein interactions imex curation rfc2_human Replication factor C subunit 2 Activator 1 subunit 2 Replication factor C 40 kDa subunit Activator 1 40 kDa subunit RFC2 protein protein human Homo sapiens Human MEVEAVCGGAGEVEAQDSDPAPAFSKAPGSAGHYELPWVEKYRPVKLNEIVGNEDTVSRLEVFAREGNVPNIIIAGPPGTGKTTSILCLARALLGPALKDAMLELNASNDRGIDVVRNKIKMFAQQKVTLPKGRHKIIILDEADSMTDGAQQALRRTMEIYSKTTRFALACNASDKIIEPIQSRCAVLRYTKLTDAQILTRLMNVIEKERVPYTDDGLEAIIFTAQGDMRQALNNLQSTFSGFGFINSENVFKVCDEPHPLLVKEMIQHCVNANIDEAYKILAHLWHLGYSPEDIIGNIFRVCKTFQMAEYLKLEFIKEIGYTHMKIAEGVNSLLQMAGLLARLCQKTMAPVAS DIP protein P35250 original sequence version: 96 B50AC8EEF89F64A9 rfc3_human Replication factor C subunit 3 RFC3 Activator 1 subunit 3 Replication factor C 38 kDa subunit Activator 1 38 kDa subunit protein protein human Homo sapiens Human MSLWVDKYRPCSLGRLDYHKEQAAQLRNLVQCGDFPHLLVYGPSGAGKKTRIMCILRELYGVGVEKLRIEHQTITTPSKKKIEISTIASNYHLEVNPSDAGNSDRVVIQEMLKTVAQSQQLETNSQRDFKVVLLTEVDKLTKDAQHALRRTMEKYMSTCRLILCCNSTSKVIPPIRSRCLAVRVPAPSIEDICHVLSTVCKKEGLNLPSQLAHRLAEKSCRNLRKALLMCEACRVQQYPFTADQEIPETDWEVYLRETANAIVSQQTPQRLLEVRGRLYELLTHCIPPEIIMKGLLSELLHNCDGQLKGEVAQMAAYYEHRLQLGSKAIYHLEAFVAKFMALYKKFMEDGLEGMMF DIP protein P40938 original sequence version: 75 820C11675A2C63A5 rfc4_human Replication factor C subunit 4 Activator 1 subunit 4 Replication factor C 37 kDa subunit Activator 1 37 kDa subunit RFC4 protein protein human Homo sapiens Human MQAFLKGTSISTKPPLTKDRGVAASAGSSGENKKAKPVPWVEKYRPKCVDEVAFQEEVVAVLKKSLEGADLPNLLFYGPPGTGKTSTILAAARELFGPELFRLRVLELNASDERGIQVVREKVKNFAQLTVSGSRSDGKPCPPFKIVILDEADSMTSAAQAALRRTMEKESKTTRFCLICNYVSRIIEPLTSRCSKFRFKPLSDKIQQQRLLDIAKKENVKISDEGIAYLVKVSEGDLRKAITFLQSATRLTGGKEITEKVITDIAGVIPAEKIDGVFAACQSGSFDKLEAVVKDLIDEGHAATQLVNQLHDVVVENNLSDKQKSIITEKLAEVDKCLADGADEHLQLISLCATVMQQLSQNC DIP protein P35249 original sequence version: 89 6FEAB3794379F3E0 rfc5_human Replication factor C subunit 5 Activator 1 subunit 5 Replication factor C 36 kDa subunit Activator 1 36 kDa subunit RFC5 protein protein human Homo sapiens Human METSALKQQEQPAATKIRNLPWVEKYRPQTLNDLISHQDILSTIQKFINEDRLPHLLLYGPPGTGKTSTILACAKQLYKDKEFGSMVLELNASDDRGIDIIRGPILSFASTRTIFKKGFKLVILDEADAMTQDAQNALRRVIEKFTENTRFCLICNYLSKIIPALQSRCTRFRFGPLTPELMVPRLEHVVEEEKVDISEDGMKALVTLSSGDMRRALNILQSTNMAFGKVTEETVYTCTGHPLKSDIANILDWMLNQDFTTAYRNITELKTLKGLALHDILTEIHLFVHRVDFPSSVRIHLLTKMADIEYRLSVGTNEKIQLSSLIAAFQVTRDLIVAEA DIP protein P40937 original sequence version: 82 E31E7B8C79933BCE rfc1_human Replication factor C subunit 1 Activator 1 subunit 1 Replication factor C large subunit Replication factor C 140 kDa subunit Activator 1 140 kDa subunit Activator 1 large subunit DNA-binding protein PO-GA RFC1 RFC140 protein protein human Homo sapiens Human MDIRKFFGVIPSGKKLVSETVKKNEKTKSDEETLKAKKGIKEIKVNSSRKEDDFKQKQPSKKKRIIYDSDSESEETLQVKNAKKPPEKLPVSSKPGKISRQDPVTYISETDEEDDFMCKKAASKSKENGRSTNSHLGTSNMKKNEENTKTKNKPLSPIKLTPTSVLDYFGTGSVQRSNKKMVASKRKELSQNTDESGLNDEAIAKQLQLDEDAELERQLHEDEEFARTLAMLDEEPKTKKARKDTEAGETFSSVQANLSKAEKHKYPHKVKTAQVSDERKSYSPRKQSKYESSKESQQHSKSSADKIGEVSSPKASSKLAIMKRKEESSYKEIEPVASKRKENAIKLKGETKTPKKTKSSPAKKESVSPEDSEKKRTNYQAYRSYLNREGPKALGSKEIPKGAENCLEGLIFVITGVLESIERDEAKSLIERYGGKVTGNVSKKTNYLVMGRDSGQSKSDKAAALGTKIIDEDGLLNLIRTMPGKKSKYEIAVETEMKKESKLERTPQKNVQGKRKISPSKKESESKKSRPTSKRDSLAKTIKKETDVFWKSLDFKEQVAEETSGDSKARNLADDSSENKVENLLWVDKYKPTSLKTIIGQQGDQSCANKLLRWLRNWQKSSSEDKKHAAKFGKFSGKDDGSSFKAALLSGPPGVGKTTTASLVCQELGYSYVELNASDTRSKSSLKAIVAESLNNTSIKGFYSNGAASSVSTKHALIMDEVDGMAGNEDRGGIQELIGLIKHTKIPIICMCNDRNHPKIRSLVHYCFDLRFQRPRVEQIKGAMMSIAFKEGLKIPPPAMNEIILGANQDIRQVLHNLSMWCARSKALTYDQAKADSHRAKKDIKMGPFDVARKVFAAGEETAHMSLVDKSDLFFHDYSIAPLFVQENYIHVKPVAAGGDMKKHLMLLSRAADSICDGDLVDSQIRSKQNWSLLPAQAIYASVLPGELMRGYMTQFPTFPSWLGKHSSTGKHDRIVQDLALHMSLRTYSSKRTVNMDYLSLLRDALVQPLTSQGVDGVQDVVALMDTYYLMKEDFENIMEISSWGGKPSPFSKLDPKVKAAFTRAYNKEAHLTPYSLQAIKASRHSTSPSLDSEYNEELNEDDSQSDEKDQDAIETDAMIKKKTKSSKPSKPEKDKEPRKGKGKSSKK 485F0332FB56819B rfc1-rfc2-2 2 3 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 6 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey association association Purification of recombinant human RFC from Sf9 cells 2A rfc1-rfc2-3 1 7 unspecified role unspecified role neutral component neutral component n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 3 unspecified role unspecified role neutral component neutral component 5 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component 4 unspecified role unspecified role neutral component neutral component physical association physical association Purification of recombinant human RFC from Sf9 cells 2B Recombinant RFC was shown to support SV40 DNA synthesis and have ATPase activity in vitro rfc1-rfc2-4 2 5 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 3 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey association association 5B When Sf9 cells were co-infected with all five RFC subunit viruses, bvHA-37kDa, bv140kDa, bv40kDa, bv38kDa, and bv36kDa, all of the untagged RFC subunits were observed to co-purify with the HA epitope-tagged 37-kDa subunit (Fig. 5A, silver-stained gels, lane 5; Fig. 5B, immunoblots of the samples in Fig. 5A probed with either an alpha -140kDa monoclonal antibody or alpha -40kDa rabbit antibody, lane 5). rfc2-rfc3-1 2 5 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 3 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey association association 5B When the 140-kDa subunit was omitted from the co-infection, a quaternary complex consisting of the 40-, 38-, 37-, and 36-kDa subunits was observed (Fig. 5A, lane 6, and Fig. 5B, alpha 40kDa immunoblot, lane 6), indicating no dependence on the 140-kDa subunit for the small subunits to assemble into a complex. rfc2-rfc4-4 2 5 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 3 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey association association 5A, 5B, However, assembly of the 140-kDa subunit into the complex was observed to be dependent upon the 38- and 40-kDa subunits, because omission of the 38-kDa subunit from the complex resulted in complete loss of the 140-kDa subunit from the complex (Fig. 5A, lane 8, and Fig. 5B, alpha 140kDa immunoblot, lane 8), rfc4-rfc3-2 2 5 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 6 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey association association 5B omission of the 40-kDa subunit resulted in significantly reduced assembly of the 140-kDa subunit into the complex (Fig. 5A, lane 7, and Fig. 5B, alpha 140kDa immunoblot, lane 7). rfc1-rfc3-1 2 3 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey association association 7 when the 36-kDa subunit was excluded from the co-infection, an interaction between the HA-140kDa subunit and the 38-, 37-, and 40-kDa subunits was observed rfc4-rfc3-3 2 5 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 4 unspecified role unspecified role prey prey physical association physical association 6 The 36- and 38-kDa subunits were found to interact directly with the 37-kDa subunit rfc3-rfc4-1 2 5 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 4 unspecified role unspecified role bait bait physical association physical association 6 The 36- and 38-kDa subunits were found to interact directly with the 37-kDa subunit rfc2-rfc4-5 2 3 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 6 unspecified role unspecified role ancillary ancillary association association 9 However, the 40-kDa subunit was discovered to interact with the 37-kDa subunit when the 36-kDa subunit was co-expressed with these two subunits (Fig. 6, rfc4-rfc5-1 2 7 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey association association 7A When either the 40- or 38-kDa subunit was omitted, only the 36- and 37-kDa subunits were found to co-immunoprecipitate with the HA-140kDa subunit (Fig. 7A rfc2-rfc3-3 1 5 unspecified role unspecified role neutral component neutral component n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 3 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component 4 unspecified role unspecified role neutral component neutral component physical association physical association Purification of recombinant human RFC from Sf9 cells 2B Recombinant RFC lacking RFC1 was shown to lack SV40 DNA synthesis but retain ATPase activity in vitro