Abstract
Arrivals of fluorescent proteins have revolutionized the way we do research in the areas of molecular cell biology. In the present study, we have successfully exploited the multimer-forming property of Red Fluorescent Protein (RFP) in living cells to preferentially shift the unliganded nuclear receptors from the nuclear to the cytoplasmic compartment. Subsequently, these cytoplasmic-shifted unliganded receptors could be induced to translocate into the nucleus by their ligands. Though the multimerization of RFP as a protein-tag is viewed as a disadvantage, we have exploited and projected this property towards novel applications in validating the clinical drugs, herbal compounds, metabolic disruptors etc. Such cytoplasmic shifted transcription factors can offer a unique opportunity to study receptor-ligand interactions and functional dynamics by analyzing ligand-mediated receptor translocation from cytoplasmic compartment to the nucleus of living cells.
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Acknowledgements
The research work presented in this study was financially supported by a research grant to RKT from the UGC (major project) and UPE-II. Financial support to our Centre by UGC-SAP, ICMR-CAR, DST-PURSE are gratefully acknowledged. AKD and ASY acknowledge DST-INSPIRE and UGC-JNU respectively for the grant of doctoral research fellowships. We apologize to those of our colleagues whose significant contributions in the subject of this research paper are not cited due to limited space. The GFP and RFP constructs generated in this study are available upon request.
We dedicate this paper to the memory Prof. Rodger Y Tsien, who was the recipient of 2008 Nobel Prize in Chemistry for his discovery and development of the fluorescent proteins.
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Dash, A.K., Yende, A.S. & Tyagi, R.K. Novel Application of Red Fluorescent Protein (DsRed-Express) for the Study of Functional Dynamics of Nuclear Receptors. J Fluoresc 27, 1225–1231 (2017). https://doi.org/10.1007/s10895-017-2109-z
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DOI: https://doi.org/10.1007/s10895-017-2109-z