# Rescue and repair of stalled ribosome damaged by ribosome-specific ribotoxins

> **NIH NIH R01** · UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN · 2023 · $59,474

## Abstract

Project Summary/Abstract
 Colicin E3 was the first ribotoxin to be characterized 50 years ago. It makes a single but precise cut of
16S rRNA in the decoding center of bacterial ribosome, resulting in stalled ribosome and eventual cell
death. Over the last half of a century, it is unclear whether there exists a biological system that is able to
reverse the ribosomal damage by colicin E3 to allow cell to survive. Employing approaches of
bioinformatics, biochemistry, structural biology, and microbiology, we have uncovered a bacterial two-
component system, RtcB and PrfH, as the antidote of colicin E3. Specifically, bacterial PrfH recognizes the
damaged and stalled ribosome and performs ribosomal rescue. This is followed by RtcB repairing the
damaged 30S ribosomal subunit. The sequential events described above are supported by abundant
preliminary data from both our in vitro and in vivo studies. In this application, we plan to significantly expand
our preliminary studies to systematically characterize the rescue and repair of bacterial ribosome with
specific damage in the decoding center with the following three main aims: (1) We will provide insight into
bacterial PrfH recognizing and rescuing the damaged and stalled 70S ribosome in vitro; (2) We will
biochemically and structurally characterize bacterial RtcB in vitro, with the emphasis of PrfH-coupled RtcB
repairing the damaged 30S ribosomal subunit; and (3) We will elucidate in vivo biological functions of RtcB-
PrfH using an in vivo attenuated RNA damage system we have developed.

## Key facts

- **NIH application ID:** 10799097
- **Project number:** 3R01GM145060-02S1
- **Recipient organization:** UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
- **Principal Investigator:** Raven H Huang
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $59,474
- **Award type:** 3
- **Project period:** 2022-05-01 → 2026-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10799097

## Citation

> US National Institutes of Health, RePORTER application 10799097, Rescue and repair of stalled ribosome damaged by ribosome-specific ribotoxins (3R01GM145060-02S1). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10799097. Licensed CC0.

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