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bioRxiv · 10.1101/2023.07.25.550078

Evaluating steroid hormone receptor interactions using the live-cell NanoBRET proximity assay

Huggins, R. J.·Hosfield, D.·Ishag-Osman, A.·Lee, K.·Ton-That, E.·Greene, G. L.
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Abstract

Steroid hormone receptors play a crucial role in the development and characterization of the majority of breast cancers. These receptors canonically function through homodimerization, but physical interactions between different hormone receptors play a key role in cell functions as well. The estrogen receptor (ER) and progesterone receptor (PR), for example, are involved in a complex set of interactions known as ER/PR crosstalk. Here, we developed a valuable panel of nuclear receptor expression plasmids specifically for use in NanoBRET assays to assess nuclear receptor homo- and heterodimerization. We demonstrate the utility of this assay system by assessing ER/PR physical interaction in the context of the endocrine therapy resistance- associated ER Y537S mutation. We identify a role of the ER Y537S mutation beyond that of constitutive activity of the receptor; it also increases ER/PR crosstalk. In total, the NanoBRET assay provides a novel avenue for investigating hormone receptor crosstalk. Future research may use this system to assess the effects of other clinically significant hormone receptor mutations on hormone receptor crosstalk.

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Huggins, R. J., Hosfield, D., Ishag-Osman, A., Lee, K., Ton-That, E., Greene, G. L.. 2023-07-26. Evaluating steroid hormone receptor interactions using the live-cell NanoBRET proximity assay. https://doi.org/10.1101/2023.07.25.550078

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