# Oxidative stress response and metabolic reprogramming by protein posttranslational arginylation

> **NIH NIH R01** · UNIVERSITY OF MIAMI SCHOOL OF MEDICINE · 2020 · $314,675

## Abstract

Oxidative stress response and metabolic reprogramming by protein posttranslational arginylation
Posttranslational arginylation is the addition of one extra arginine to a protein. This modification often leads to
rapid protein degradation. In fungi and animals, arginylation is solely mediated by Arginyltransferase1 (ATE1).
Multiple lines of evidence from our group and others have shown that ATE1 and its arginylation activity are
essential for cellular response to a variety of oxidative stressors and the associated metabolic reprogramming
including glycolysis. However, the molecular mechanisms by which arginylation regulate oxidative stress
response (OSR) and metabolism remain unknown. This gap of knowledge makes it difficult to devise
approaches to intervene in arginylation for the prevention or treatment of diseases such as inflammation,
cardiovascular abnormalities, cancer, and aging-related maladies, which are often derived from dysregulated
OSR and the associated metabolic alterations.
Following from our recent studies where we showed that ATE1 and arginylation activity are increased in cells
under acute oxidative stress, and downregulated upon chronic exposures to stressors, we aim to understand
exactly how arginylation influences OSR. The lack of understanding for arginylation is largely due to major
technical challenges in the field for identifying the majority of arginylation substrates, which degrade rapidly. To
overcome this problem, we redesign a new approach combining indirect methods to identify the impact of
arginylation on protein stability and direct methods to identify arginylation modification on proteins. The power
of this new approach was demonstrated in our preliminary screening, in which we identified several new
arginylation candidates including hypoxia-inducible factor 1 (HIF1), a critical regulator of OSR, glycolysis,
and mitochondrial respiration. Our data further suggested that the functional role of HIF1a is regulated by
arginylation. Following from this breakthrough, the objective of this proposed study is to reveal how arginylation
regulates oxidative stress response and associated metabolic reprogramming by affecting critical proteins
including HIF1. We will apply and expand our newly developed approach to identify additional arginylated
proteins (Aim1), elucidate the effects of arginylation on key substrates including HIF1 in regulating stress
response/metabolism (Aim 2), and using high-throughput methods to characterize the global impact of
arginylation on different cellular pathways (Aim 3).
In the end of this study, we are expected to reveal major molecular mechanisms of how posttranslational
arginylation of HIF1 and other critical proteins regulates OSR and metabolism. We would also uncover global
impacts of arginylation, which is mediated by a single enzyme ATE1, in many cellular pathways. A long-lasting
impact is further anticipated by the discoveries of many new and unexpected targets of arginylation.

## Key facts

- **NIH application ID:** 10030477
- **Project number:** 1R01GM138557-01
- **Recipient organization:** UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
- **Principal Investigator:** FANGLIANG ZHANG
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $314,675
- **Award type:** 1
- **Project period:** 2020-09-01 → 2024-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10030477

## Citation

> US National Institutes of Health, RePORTER application 10030477, Oxidative stress response and metabolic reprogramming by protein posttranslational arginylation (1R01GM138557-01). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10030477. Licensed CC0.

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