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Real-time analysis of ribosomal frameshifting and its impact on immunity and disease

Project description

Exploring ribosomal frameshifting in human cells

Many viruses, including HIV, employ a process known as programmed ribosomal frameshifting (PRF) where, during protein synthesis, the ribosome slips to another position. This process alters the code and leads to the generation of different proteins from the same mRNA, adding another level of gene expression regulation. The EU-funded T-FRAME project aims to study the phenomenon of PRF in human cells and identify the factors that regulate it. Scientists will employ human T lymphocytes and HIV-1 as a model system to investigate the impact of PRF on infection and innate immunity. Results will provide unprecedented knowledge on this alternative pathway to standard decoding and pave the way for the design of novel antiviral therapies.

Objective

The coding region of many genes contains sequence elements that constitute roadblocks during mRNA translation. These roadblocks present problems, but also opportunities for the cell to increase the coding capacity of the genome by so-called programmed ribosome frameshifting (PRF). PRF is a universal gene expression strategy that brings plasticity to the proteome. It is widely studied in viruses and bacteria, yet poorly understood in humans.

My own mechanistic discoveries have highlighted that, regardless of various stimulatory structures and slippery nucleotides involved, PRF in bacterial and viral mRNAs follows one of the two general pathways depending on substrate. In contrast, many questions remain regarding the occurrences of PRF in humans and its regulation by recently discovered trans-acting viral and cellular factors.

In T-FRAME, I will investigate the mechanism of PRF and its role in disease using HIV infection of human-T lymphocytes as an in vivo paradigm. To study molecular details of how trans-factors modulate PRF, I will leverage cutting-edge ensemble and single-molecule techniques to monitor the ribosome as it moves along the mRNA codon per codon. In addition, I will assess mRNA interactions with PRF factors during this process. Using a PRF-RNA-interactome capture assay that we have recently developed, I will search for new trans- factors in human cells. Finally, I will study the translation of T cells and HIV-1 on a global level during infection to understand the full scope and dynamic nature of PRF in this model system.

To summarize, T-FRAME will advance our understanding of how RNA structure and trans-factors shape translation regimes in higher eukaryotes and how deviations from the standard decoding path impact infection and innate immunity. I envision harnessing these findings to develop novel tools for synthetic biology and new design principles for RNA-centric antiviral and immune therapies.

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ERC-STG - Starting Grant

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Call for proposal

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(opens in new window) ERC-2020-STG

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Host institution

UNIVERSITAET REGENSBURG
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.

€ 874 370,50
Address
UNIVERSITATSSTRASSE 31
93053 Regensburg
Germany

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Region
Bayern Oberpfalz Regensburg, 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.

€ 874 370,50

Beneficiaries (2)

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