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Regulation of mRNA homeostasis by Poly(A) binding proteins

Project description

Investigating PABPCs in RNA regulation and homeostasis

RNA is essential for life, and cells maintain its concentration (RNA homeostasis) by balancing synthesis and degradation. A feedback mechanism may coordinate mRNA levels through a ‘sensor’ that communicates mRNA concentrations to transcription and degradation controls. Cytoplasmic poly(A)-binding proteins (PABPCs) bind to mRNAs to regulate their stability and translation, but their role in maintaining RNA homeostasis under physiological conditions remains unclear. Supported by the Marie Skłodowska-Curie Actions programme, the RNAhome project will use PABPC titration, metabolic labelling, and multiplexed imaging to analyse RNA dynamics in response to varying PABPC levels and localisation, aiming to uncover fundamental mechanisms of RNA homeostasis and broader principles of gene regulation.

Objective

RNA is a biomolecule central to all life forms, and cells regulate RNA production and degradation rates to maintain stable RNA concentrations (RNA homeostasis) through a largely unexplored mechanism. Increasing evidence indicates that a feedback mechanism coordinates mRNA synthesis and degradation, allowing cells to buffer mRNA concentration. For the coordination of these processes, I hypothesise the existence of a “sensor” that “measures” mRNA concentration and conveys this information to the transcription and/or degradation machinery, establishing a coordinated response.

Cytoplasmic Poly(A)-binding proteins (PABPCs) are highly conserved across eukaryotes, and specifically bind to polyadenylated mRNAs, regulating their stability, translation, and degradation. Several properties of PABPCs, such as nuclear shuttling and transcription inhibition in response to viral infection, make them ideal candidates as mRNA sensors, yet it is unclear whether they control mRNA homeostasis in physiological conditions.

RNAhome will characterise the role of PABPCs in mRNA buffering and homeostasis using a systemic approach. I will conduct PABPC titration, followed by metabolic labelling and multiplexed imaging, to analyse RNA dynamics in response to varying PABPC expression and localisation. I will use my extensive expertise in nanoinjection to introduce mRNA into live cells and assess if PABPCs can directly sense mRNA levels and modulate transcription/turnover accordingly. Finally, I will use SLAMseq to measure genome-wide RNA dynamics with transcript-level resolution, establishing the impact of PABPC expression on mRNA dynamics.

RNAhome is a prime example of interdisciplinary science, combining the researcher’s expertise in bio-nanotechnology and physical sciences with RNA dynamics knowledge of the host group, and world-class facilities of the host organisation to unravel the fundamental mechanisms governing RNA homeostasis and drive innovation in RNA-based technologies.

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HORIZON-TMA-MSCA-PF-EF - HORIZON TMA MSCA Postdoctoral Fellowships - European Fellowships

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(opens in new window) HORIZON-MSCA-2024-PF-01

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Coordinator

FONDAZIONE HUMAN TECHNOPOLE
Net EU contribution

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€ 193 643,28
Address
VIALE LEVI MONTALCINI RITA 1
20157 Milano
Italy

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Region
Nord-Ovest Lombardia Milano
Activity type
Research Organisations
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