# Imaging Protein Synthesis on the Ribosome using Single-Molecule FRET

> **NIH NIH R01** · ST. JUDE CHILDREN'S RESEARCH HOSPITAL · 2020 · $500,813

## Abstract

PROJECT ABSTRACT:
The mechanism of protein synthesis and its regulation in the cell determines the diversity and capacity of the
proteome. The central integration point for this regulatory control is the ribosome: a two-subunit, megadalton
RNA-protein assembly. Highlighting the exquisite sensitivity of translation and the ribosome to regulation, the
majority of known antibiotics either dysregulate or block ribosome function. Correspondingly, delineation of the
protein synthesis mechanism in molecular detail has the potential to inform on paradigms of gene expression
control and on how to combat the global health threat of emerging and drug resistant pathogens. As the loss of
translation control is a hallmark of cancer, a deeper understanding of the protein synthesis mechanism also
holds the promise of targeted therapeutic strategies for human disease treatments that are currently lacking.
Investigations into structure-function relationships governing the translation mechanism have been principally
conducted in bacteria using traditional ensemble methods. Such studies have revealed that the phase of
translation in which protein is synthesized from messenger RNA (mRNA), termed elongation, is the most time
intensive and commonly drug-targeted. They have also discerned that elongation entails the ribosome transiently
interacting with specific cellular components through an ordered series of events, where the decoding of each
mRNA codon is accompanied by large-scale conformational changes within the ribosome and interacting factors,
and between the ribosome and its mRNA and transfer RNA (tRNA) substrates. The need for large amounts of
homogenous material has thwarted analogous investigations of the human translation mechanism. Hence,
conserved and divergent features of the translation mechanism between single-cell organisms and mammals
that determine the molecular basis of antibiotic specificity have remained largely obscure. Here, we seek to
delineate common and distinct features of bacterial and human protein synthesis — and the translation
mechanisms in healthy and cancerous human cells — to: 1] improve the efficacies of existing antibiotics; 2]
develop new strategies for antibiotic interventions; and 3] explore the possibility of therapies targeting unchecked
proliferative cell growth and metastatic spread. We will do so by establishing quantitative, structural and kinetic
frameworks for the elemental steps of elongation in bacteria and humans using an integrated battery of
biophysical methods, including single-molecule fluorescence imaging and state-of-the-art cryo-electron
microscopy. Our collaborative investigations will delineate the order and timing of conformational events
underpinning fidelity in bacterial and human elongation cycles and the structural and mechanistic distinctions
that determine the efficacies of clinically relevant antibiotics targeting these processes. These insights will shed
light on how translation control is achieved,...

## Key facts

- **NIH application ID:** 10072525
- **Project number:** 2R01GM079238-15
- **Recipient organization:** ST. JUDE CHILDREN'S RESEARCH HOSPITAL
- **Principal Investigator:** Scott C Blanchard
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $500,813
- **Award type:** 2
- **Project period:** 2006-09-15 → 2024-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10072525

## Citation

> US National Institutes of Health, RePORTER application 10072525, Imaging Protein Synthesis on the Ribosome using Single-Molecule FRET (2R01GM079238-15). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10072525. Licensed CC0.

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