# Translational Control by 5'-untranslated regions

> **NIH NIH R01** · YALE UNIVERSITY · 2021 · $331,220

## Abstract

PROJECT SUMMARY
Significance: Translation initiation is a highly regulated step in eukaryotic gene expression and
its dysregulation is linked to heritable human diseases and cancer. Translational regulation
depends on cellular condition-dependent differences in the protein output of mRNAs, but the
key mRNA features that distinguish efficiently translated mRNAs are largely unknown. A
predictive understanding of these mRNA characteristics is needed to harness the broad
therapeutic potential of drugs that target translation factors, develop new translation-based
therapies, and engineer therapeutic mRNAs.
Approach: Our work aims to comprehensively identify 5′-UTR sequences that enhance or
repress translation and illuminate their underlying mechanisms. We hypothesize that 5′-UTRs
with unexplained high or low translation activity bind proteins that function as mRNA-specific
translational activators or repressors. We envision two broad categories of translational
enhancers: 5′-UTR sequences or RNA structures that bind to core initiation factors
preferentially, and 5′-UTR elements that bind novel factors whose roles in ribosome recruitment
remain to be elucidated. In support of the first model, we have recently demonstrated strong
sequence preferences for yeast eIF4G1 and found that its preferred binding motif, oligo-uridine,
occurs naturally in the 5′-UTRs of hundreds of genes and enhances their translation. Aim 1 will
leverage a new method developed in our laboratory that allows highly parallel dissection of
candidate cis regulatory elements to define the contribution of specific 5′-UTR features to
ribosome recruitment activity in cell lysates. We use S. cerevisiae (budding yeast) because 5′-
UTR isoforms are experimentally well defined in this organism and we can exploit a wealth of
genetic, biochemical and structural data. Aim 2 will reveal how much of the observed variance
in ribosome recruitment is directly explained by differences in affinities for core initiation factors.
Aim 3 will identify novel factors that bind 5′-UTR enhancer and silencer elements using
crosslinking and mass spectrometry; investigate their mechanisms that alter ribosome
recruitment in vitro; and explore their impact on regulated translation in vivo. Together, this work
will reveal how differences in mRNA primary sequence lead to large and regulated differences
in translation. Because the eukaryotic translational machinery and regulatory paradigms are
highly conserved, the molecular insights gained here are likely to be broadly illuminating for
understanding translational control, including in pathophysiological processes in humans.

## Key facts

- **NIH application ID:** 10223370
- **Project number:** 5R01GM132358-03
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** Wendy Victoria Gilbert
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $331,220
- **Award type:** 5
- **Project period:** 2019-09-17 → 2023-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10223370

## Citation

> US National Institutes of Health, RePORTER application 10223370, Translational Control by 5'-untranslated regions (5R01GM132358-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10223370. Licensed CC0.

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