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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Publication Date
2016-06-21Journal Title
Journal of Structural Biology
ISSN
1047-8477
Publisher
Elsevier
Language
English
Type
Article
This Version
AM
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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
Abstract
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.
Keywords
nuclear transport, hepatocyte nuclear factor 1β (HNF1β), nuclear localization sequence
Sponsorship
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.
Funder references
Royal Society (WM150022)
European Research Council (279337)
EPSRC (EP/J016012/1)
EPSRC (EP/K039520/1)
Cancer Research UK (C37096/A13001)
Cancer Research UK (CRUK-A15601)
Identifiers
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
Rights
Attribution-NonCommercial-NoDerivatives 4.0 International, Attribution-NonCommercial-NoDerivatives 4.0 International, Attribution-NonCommercial-NoDerivatives 4.0 International
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