Surface Plasmon Resonance Study of Cooperative Interactions of Estrogen Receptor α and Specificity Protein 1 with Composite DNA Elements

Part of the Methods in Molecular Biology book series (MIMB, volume 1366)

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.

Key words

Transcription factor Estrogenreceptor Sp1 Cooperative protein–DNA interaction Surface plasmon resonance Biacore T200 

Notes

Acknowledgements

The author would like to acknowledge the Joint Council Office of the Agency for Science, Technology and Research (A*STAR), Singapore, for financial support.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Institute of Materials Research and EngineeringAgency for Science, Technology and Research (A*STAR)InnovisSingapore

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