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All rights reserved. http://resource.belframework.org/belframework/20131211/knowledge/large_corpus.bel http://purl.org/dc/elements/1.1/title BEL Framework Large Corpus Document http://resource.belframework.org/belframework/20131211/knowledge/large_corpus.bel http://purl.org/pav/authoredBy http://www.tkuhn.ch/bel2nanopub/RA8t7clIap7MGxaB8UpZgQRKK0KyxnkwzQvRskk3_-_xY#_6 http://resource.belframework.org/belframework/20131211/knowledge/large_corpus.bel http://purl.org/pav/version 20131211 http://www.tkuhn.ch/bel2nanopub/RA8t7clIap7MGxaB8UpZgQRKK0KyxnkwzQvRskk3_-_xY#_5 http://www.w3.org/ns/prov#value Next, we wanted to test whether E2F6 binds directly to the predicted E2F-binding sites of the SMC1{beta} and STAG3 promoters. To address this question we performed gel retardation experiments with in vitro translated E2F6 and DP2 and radiolabeled oligonucleotides corresponding to the E2F sites of the SMC1{beta} and STAG3 promoters. Binding of E2F6 to these elements was compared with binding to the E2F-site of the DHFR promoter, a bona fide E2F-binding sequence (22). As shown in Fig. 3A, binding of E2F6 to the SMC1{beta} site was readily detected, whereas under the same conditions E2F6 did not significantly bind to the DHFR sequence (compare lane 1 and lane 7 in Fig. 3A). Preferential binding of E2F6 to the SMC1{beta} sites was confirmed by competition with unlabeled oligonucleotides. The SMC1{beta}, but not the DHFR oligonucleotide, efficiently competed for the binding of E2F6 to the E2F element (Fig. 3A, lanes 2–4). We concluded that E2F6 binds with higher affinity to the SMC1{beta} promoter than to the classical E2F binding site in the DHFR promoter. As described above, the STAG3 promoter contains an E2F site that is identical to the one in the SMC1{beta} promoter (Fig. 2D), and in vitro translated E2F6 readily bound to the STAG3 element (Fig. 3C).Endogenous E2F6 associated with the STAG3 promoter in wild-type MEFs (Fig. 4A, lane 3), although, as expected, its binding was weaker than the binding of the overexpressed protein. http://www.tkuhn.ch/bel2nanopub/RA8t7clIap7MGxaB8UpZgQRKK0KyxnkwzQvRskk3_-_xY#_5 http://www.w3.org/ns/prov#wasQuotedFrom http://www.ncbi.nlm.nih.gov/pubmed/16236716 http://www.tkuhn.ch/bel2nanopub/RA8t7clIap7MGxaB8UpZgQRKK0KyxnkwzQvRskk3_-_xY#_6 http://www.w3.org/2000/01/rdf-schema#label Selventa http://www.tkuhn.ch/bel2nanopub/RA8t7clIap7MGxaB8UpZgQRKK0KyxnkwzQvRskk3_-_xY#assertion http://www.w3.org/ns/prov#hadPrimarySource http://www.ncbi.nlm.nih.gov/pubmed/16236716 http://www.tkuhn.ch/bel2nanopub/RA8t7clIap7MGxaB8UpZgQRKK0KyxnkwzQvRskk3_-_xY#assertion http://www.w3.org/ns/prov#wasDerivedFrom http://resource.belframework.org/belframework/20131211/knowledge/large_corpus.bel http://www.tkuhn.ch/bel2nanopub/RA8t7clIap7MGxaB8UpZgQRKK0KyxnkwzQvRskk3_-_xY#assertion http://www.w3.org/ns/prov#wasDerivedFrom http://www.tkuhn.ch/bel2nanopub/RA8t7clIap7MGxaB8UpZgQRKK0KyxnkwzQvRskk3_-_xY#_5 http://www.tkuhn.ch/bel2nanopub/RA8t7clIap7MGxaB8UpZgQRKK0KyxnkwzQvRskk3_-_xY#pubinfo http://www.tkuhn.ch/bel2nanopub/RA8t7clIap7MGxaB8UpZgQRKK0KyxnkwzQvRskk3_-_xY http://purl.org/dc/terms/created 2014-07-03T14:32:51.587+02:00 http://www.tkuhn.ch/bel2nanopub/RA8t7clIap7MGxaB8UpZgQRKK0KyxnkwzQvRskk3_-_xY http://purl.org/pav/createdBy http://orcid.org/0000-0001-6818-334X http://www.tkuhn.ch/bel2nanopub/RA8t7clIap7MGxaB8UpZgQRKK0KyxnkwzQvRskk3_-_xY http://purl.org/pav/createdBy http://orcid.org/0000-0002-1267-0234