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Structural and calorimetric studies demonstrate that the hepatocyte nuclear factor 1β (HNF1β) transcription factor is imported into the nucleus via a monopartite NLS sequence.

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Wiedmann, Mareike M 
Aibara, Shintaro 
Spring, David R 
Stewart, Murray 
Brenton, James D 


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. Pull-down 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 (229)KKMRRNR(235) 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 2.4Å 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 provide the basis for the development of compounds targeting transcription factor HNF1β via its nuclear import pathway.



Hepatocyte nuclear factor-1β (HNF1β), Importin-α, Isothermal titration calorimetry, Nuclear import pathway, Nuclear localisation signal sequence (NLS), Site-directed mutagenesis, X-ray crystallography, Active Transport, Cell Nucleus, Animals, Binding Sites, Cell Line, Tumor, Cell Nucleus, Crystallography, X-Ray, HEK293 Cells, Hepatocyte Nuclear Factor 1-beta, Humans, Mice, Models, Molecular, Nuclear Localization Signals, Protein Binding, Protein Conformation, alpha-Helical, Sequence Analysis, Protein, Xenopus Proteins, Xenopus laevis, alpha Karyopherins

Journal Title

J Struct Biol

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Elsevier BV
Royal Society (WM150022)
European Research Council (279337)
Engineering and Physical Sciences Research Council (EP/J016012/1)
Engineering and Physical Sciences Research Council (EP/K039520/1)
Cancer Research UK (C37096/A13001)
Cancer Research UK (15601)
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.