title: Surface Plasmon Resonance Study of Cooperative Interactions of Estrogen Receptor α and Specificity Protein 1 with Composite DNA Elements.
abstract: Estrogen receptor α (ERα) and Specificity protein 1 (Sp1) are transcription factors (TF) that are involved in regulating progesterone receptor (PR) gene expression through cooperative interactions with DNA. The natural composite DNA +571 ERE/Sp1 site in promoter A of the progesterone receptor contains a half-site of estrogen response elements (½ERE) upstream of two Sp1 binding sites (the proximal Sp1 (Sp1/P) and distal Sp1 (Sp1/D)) with a 4 bp spacer. Here, we have developed a protocol for studying the cooperative interaction of Sp1 and ERα with the composite DNA of +571 ERE/Sp1 site using Biacore T200, a high sensitivity surface plasmon resonance spectroscopy. With this protocol, we have concluded that Sp1 binding enhances the overall ERα binding to the composite DNA. We have also determined the optimal spacer distance between the ½ERE and Sp1/D for the best cooperative protein binding. This study is pivotal in guiding the bioinformatics simulation to yield an exact model of the spacer dependency of the transcription factor/cofactor-DNA interactions, which is important for understanding the nuclear receptor regulating activity through other coactivators.
authors.affiliation: Institute of Materials Research and Engineering, Agency for Science, Technology and Research (A*STAR), 2 Fusionopolis Drive, #08-03, Innovis, Singapore, 138632. xd-su@imre.a-star.edu.sg.
authors.forename: Xiaodi
authors.initials: X
authors.lastname: Su
authors.affiliation: Institute of Materials Research and Engineering, Agency for Science, Technology and Research (A*STAR), 2 Fusionopolis Drive, #08-03, Innovis, Singapore, 138632.
authors.forename: Hong Yan
authors.initials: HY
authors.lastname: Song
mesh_terms: D001665:Binding Sites; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D011485:Protein Binding; D011980:Receptors, Progesterone / Q000235:genetics* / Q000378:metabolism; D020218:Response Elements; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D020349:Surface Plasmon Resonance / Q000379:methods*; D057188:Workflow
chemical_list: C506487:ESR1 protein, human; D047628:Estrogen Receptor alpha; D011980:Receptors, Progesterone; D016329:Sp1 Transcription Factor; C108209:SP1 protein, human
keywords: Biacore T200; Cooperative protein–DNA interaction; Estrogen receptor; Sp1; Surface plasmon resonance; Transcription factor