Structural and calorimetric studies demonstrate that the hepatocyte nuclear factor 1β (HNF1β) transcription factor is imported into the nucleus via a monopartite NLS sequence
| Autor: | Mareike M. Wiedmann, Murray Stewart, Shintaro Aibara, James D. Brenton, David R. Spring |
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| Přispěvatelé: | Spring, David [0000-0001-7355-2824], Brenton, James [0000-0002-5738-6683], Apollo - University of Cambridge Repository |
| Rok vydání: | 2016 |
| Předmět: |
Models
Molecular Protein Conformation alpha-Helical 0301 basic medicine mImportin-α mouse Importin-α Nuclear Localization Signals Nuclear import pathway Xenopus Proteins Crystallography X-Ray environment and public health GST glutathione S-transferase HNF1βDBD HNF1β DNA binding domain Mice Xenopus laevis 0302 clinical medicine Sequence Analysis Protein Structural Biology Nuclear protein CCC ovarian clear cell carcinoma IBB Importin-β-binding domain Hepatocyte nuclear factor-1β (HNF1β) Nuclear localisation signal sequence (NLS) 3. Good health Cell biology medicine.anatomical_structure 030220 oncology & carcinogenesis Isothermal titration calorimetry Protein Binding alpha Karyopherins Active Transport Cell Nucleus Importin Biology Article HNF1β hepatocyte nuclear factor 1β 03 medical and health sciences Importin-α Cell Line Tumor medicine Animals Humans Transcription factor Hepatocyte Nuclear Factor 1-beta X-ray crystallography Cell Nucleus Site-directed mutagenesis Binding Sites xImportin-α Xenopus Importin-α DNA-binding domain Molecular biology Cell nucleus HEK293 Cells 030104 developmental biology NLS nuclear localization signal Hepatocyte Nuclear Factor 1-Beta Nuclear transport Nuclear localization sequence |
| Zdroj: | Journal of Structural Biology |
| ISSN: | 1047-8477 |
| DOI: | 10.1016/j.jsb.2016.06.018 |
| Popis: | 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 2.4Å 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. |
| Databáze: | OpenAIRE |
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