# Viral Nucleic Acid Stimulation of Translation

> **NIH NIH R21** · UNIVERSITY OF TEXAS AT AUSTIN · 2020 · $189,443

## Abstract

Project Summary
 Innate immune responses involve the detection of foreign nucleic acids, including those from
viruses. Multiple protein sensors detect the presence of viral RNA and DNA, leading to type I interferon
(IFN) signaling and translational suppression of capped RNAs. Nevertheless, pathogens thwart this
response by various mechanisms. Our published results revealed that unintegrated DNA containing
specific retroviral sequences leads to increased cap-dependent translation of co-introduced genes (termed
superinduction). High levels of lentiviral, gammaretroviral, and betaretroviral DNA resulted in superinduction
in rat, mouse, and human cell lines using multiple transfection methods. Surprisingly, no differences were
observed in type I IFN or mTOR signaling as well as the stress, DNA damage, or unfolded protein
responses in the presence of retroviral sequences relative to standard plasmid vectors. These results led to
the hypothesis that high levels of unintegrated retroviral DNA are detected by a factor that signals to the
cap-binding complex to increase translation of capped RNAs from newly introduced DNAs. In the first
specific aim, different parts of the superinduction pathway will be identified. An antibody-based screen that
detects differences in phosphorylation in multiple signal transduction pathways will be used to detect
signaling differences during superinduction. Protein-protein interactions with the translation initiation factor
eIF-4E, which increases during superinduction, or other translation-related factors detected in the screen
will be used to covalently trap and identify other signaling partners using a newly developed method,
Ubiquitin-Activated Interaction Traps or UBAITs. Trapped proteins then will be purified and identified by
mass spectroscopy. In the second specific aim, optimal sequences for superinduction will be defined by
testing both engineered and synthesized retroviral and non-retroviral sequences after transduction or
transfection. Using an optimized superinducing sequence compared to a non-inducing sequence,
biotinylated DNAs will be introduced into cells, and the associated proteins will be purified using streptavidin
beads and identified by mass spectrometry. The goal of these studies is to understand how mammalian
cells respond to the presence of foreign DNA. This proposal has immediate short-term benefits for
optimization of DNA transfection for protein expression as well as for retroviral transduction experiments.
Long-term benefits include identification of the signaling pathways for detection of foreign nucleic acids as
well as therapeutic targets with applications for gene therapy and anti-viral drugs.

## Key facts

- **NIH application ID:** 9830012
- **Project number:** 5R21AI142418-02
- **Recipient organization:** UNIVERSITY OF TEXAS AT AUSTIN
- **Principal Investigator:** Jaquelin Page Dudley
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $189,443
- **Award type:** 5
- **Project period:** 2018-12-01 → 2021-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9830012

## Citation

> US National Institutes of Health, RePORTER application 9830012, Viral Nucleic Acid Stimulation of Translation (5R21AI142418-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9830012. Licensed CC0.

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