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Molecular, Structural, and Functional Studies of Leukemia-Associated Transcription Factor-Nucleoporin Fusion Proteins

Project description

Study of leukaemia-associated nucleoporin fusion proteins

Nuclear pores enable the passive and facilitated transport of molecules across the nuclear envelope, and nucleoporins are the main components of the nuclear pore complexes in eukaryotic cells. Nup98 is a mobile nucleoporin that is present at the nuclear pore complex and within the nucleus. Nup98-HoxA9 (NHA9), a fusion between the phenylalanine–glycine-rich region of Nup98 and the homeobox transcription factor HoxA9, is one of the most frequent Nup98 fusions associated with acute myeloid leukaemia. The EU-funded TFNup project aims to develop a technological platform combining chemical biology, microfluidics and high-resolution molecular imaging to study the structure and function of NHA9 in vitro and in the cells in order to define a new pathway role in gene dysregulation.

Objective

Nup98 is a mobile nucleoporin that localizes both at the nuclear pore complex and within the nucleus. Nup98 is frequently rearranged to form leukemogenic Nup98-fusion proteins with various partners. Nup98-HoxA9 (NHA9), a fusion between phenylalanine-glycine-rich (FG-rich) region of Nup98 and the homeobox transcription factor (TF) HoxA9, is one of the most frequent Nup98-fusion associated with acute myeloid leukemia. The physiological role of NHA9 in hematopoietic development has been gradually established in the past decade at the cellular level. However, the plasticity and the phase separation behavior of such intrinsically disordered proteins (IDPs) largely hinder our understanding of their functions in gene regulation at the molecular level. In this project, I will develop platform technologies that combine chemical biology, microfluidics, and high-resolved molecular imaging to study the structure and biophysical function of NHA9 in vitro and in cells, and open up a new pathway to unravel its role in gene dysregulation from molecular perspective. Firstly, I will dual-label NHA9 at specific sites using the cutting-edge genetic code expansion technology developed by the host laboratory, and characterize the plasticity of NHA9 in live cells using Fluorescence lifetime imaging (FLIM) based Förster resonance energy transfer (FRET) platform. Next, I will use my strengths in microfluidics to design a new platform for tracking the phase separation behaviors of NHA9 with high temporal resolution in vitro. Finally, I will fuse NHA9 with proximity-dependent biotin identification (BioID) tags, and visualize the dynamic interaction networks of NHA9 using super-resolution microscopy (SRM) in live cells. By integrating those interdisciplinary approaches, the proposed research would make a conceptual breakthrough in understanding the molecular mechanism of NHA9-driven leukemogenesis and may provide a rationale for the search of potential therapeutic approaches in the future.

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MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)

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Call for proposal

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(opens in new window) H2020-MSCA-IF-2019

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Coordinator

JOHANNES GUTENBERG-UNIVERSITAT MAINZ
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 174 806,40
Address
SAARSTRASSE 21
55122 MAINZ
Germany

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Region
Rheinland-Pfalz Rheinhessen-Pfalz Mainz, Kreisfreie Stadt
Activity type
Higher or Secondary Education Establishments
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Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

€ 174 806,40
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