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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

DOI: 10.1016/j.jsb.2016.06.018 DOI Help

Authors: Mareike M. Wiedmann (Department of Chemistry, University of Cambridge; Cancer Research UK Cambridge Institute, University of Cambridge) , Shintaro Aibara (MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Diamond Light Source) , David R. Spring (Department of Chemistry, University of Cambridge) , Murray Stewart (MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus) , James D. Brenton (Cancer Research UK Cambridge Institute, University of Cambridge)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of Structural Biology , VOL 195 , PAGES 273 - 281

State: Published (Approved)
Published: September 2016

Open Access Open Access

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. 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 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 provide the basis for the development of compounds targeting transcription factor HNF1β via its nuclear import pathway.

Journal Keywords: Importin-α; Nuclear import pathway; Nuclear localisation signal sequence (NLS); Hepatocyte nuclear factor-1β (HNF1β); Site-directed mutagenesis; X-ray crystallography; Isothermal titration calorimetry

Subject Areas: Biology and Bio-materials, Medicine, Chemistry

Instruments: I03-Macromolecular Crystallography

Added On: 26/08/2016 16:24

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