Structural and calorimetric studies demonstrate that the Hepatocyte Nuclear Factor 1β (HNF1β) transcription factor is imported into the nucleus via a monopartite NLS sequence
Journal of Structural Biology
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Wiedmann, M. M., Aibara, S., Spring, D., Stewart, M., & Brenton, J. (2016). Structural and calorimetric studies demonstrate that the Hepatocyte Nuclear Factor 1β (HNF1β) transcription factor is imported into the nucleus via a monopartite NLS sequence. Journal of Structural Biology https://doi.org/10.1016/j.jsb.2016.06.018
The transcription factor hepatocyte nuclear factor 1β (HNF1β) is ubiquitously overexpressed in ovarian clear cell carcinoma (CCC) and is a potential therapeutic target. To explore potential approaches that block HNF1β transcription we have identified and characterised extensively the nuclear localisation signal (NLS) for HNF1β and its interactions with the nuclear protein import receptor, Importin-α. Pulldown assays demonstrated that the DNA binding domain of HNF1β interacted with a spectrum of Importin-α isoforms and deletion constructs tagged with eGFP confirmed that the HNF1β 229KKMRRNR235 sequence was essential for nuclear localisation. We further characterised the interaction between the NLS and Importin-α using complementary biophysical techniques and have determined the 2.4 Å resolution crystal structure of the HNF1β NLS peptide bound to Importin-α. The functional, biochemical, and structural characterisation of the nuclear localisation signal present on HNF1β and its interaction with the nuclear import protein Importin-α provides the basis for the development of compounds targeting transcription factor HNF1β via its nuclear import pathway.
nuclear transport, hepatocyte nuclear factor 1β (HNF1β), nuclear localization sequence
We thank our colleagues in Cambridge for their assistance, comments and criticisms. M.W. is funded by Cancer Research UK, Department of Chemistry at the University of Cambridge, School of the Physical Sciences and the Cambridge Cancer Centre. Funding in part was also provided by Medical Research Council Grant U105178939 to M.S. We would like to thank the Biorepository, Research Instrumentation, and Microscopy facilities at the Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK for assistance and Matthew Maggiolini for proofreading. We are grateful for the use of the Diamond Light Source Synchrotron (Harwell Science & Innovation Campus, Didcot, OX11 0DE, UK) for data collection.
Royal Society (WM150022)
European Research Council (279337)
Cancer Research UK (C37096/A13001)
Cancer Research UK (CRUK-A15601)
External DOI: https://doi.org/10.1016/j.jsb.2016.06.018
This record's URL: https://www.repository.cam.ac.uk/handle/1810/256876
Attribution-NonCommercial-NoDerivatives 4.0 International, Attribution-NonCommercial-NoDerivatives 4.0 International, Attribution-NonCommercial-NoDerivatives 4.0 International
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