PT  - JOURNAL ARTICLE
AU  - Mahmood Hassan Dalhat
AU  - Hani Choudhry
AU  - Mohammad Imran Khan
TI  - NAT10, an RNA acetyl cytidine transferase restrains ferroptosis in cancer cells by maintaining SLC7A11 RNA stability
AID  - 10.1101/2022.11.22.517547
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.11.22.517547
4099  - http://biorxiv.org/content/early/2022/11/24/2022.11.22.517547.short
4100  - http://biorxiv.org/content/early/2022/11/24/2022.11.22.517547.full
AB  - Recently, we reported that N-acetyltransferase 10 (NAT10) regulates fatty acid metabolism through ac4C-dependent RNA modification of key genes in cancer cells. During this work, we noticed ferroptosis as one of the most negatively enriched pathways among other pathways in NAT10 depleted cancer cells. In the current work, we explored the possibility of whether NAT10 acts as an epitrancriptomic regulator of ferroptosis pathway in cancer cells. Global ac4C levels and expression of NAT10 with other ferroptosis-related genes were assessed via dotblot and RT-qPCR respectively. Flow cytometry and biochemical analysis were used to assess oxidative stress and ferroptosis features. The ac4C mediated mRNA stability was conducted RIP-PCR and mRNA stability assay. Metabolites were profiled using LC-MS/MS. Our results showed significant downregulation in expression of essential genes related to ferroptosis namely SLC7A11, GCLC, MAP1LC3A, and SLC39A8 in NAT10 depleted cancer cells. Further, we noticed a reduction in cystine uptake and reduced GSH levels along with elevated ROS, and lipid peroxidation levels in NAT10 depleted cells. Consistently, overproduction of oxPLs as well as increased mitochondrial depolarization and decreased activities of antioxidant enzymes support the notion of ferroptosis induction in NAT10 depleted cancer cells. Mechanistically, reduced ac4C level shortens the half-life of GCLC and SLC7A11 mRNA, resulting in low levels of intracellular cystine and reduced GSH, failing to detoxify ROS leading to increased cellular oxPLs which facilitates ferroptosis induction. Collectively, our findings suggest that NAT10 restrains ferroptosis by stabilizing the SLC7A11 mRNA transcripts to avoid oxidative stress that induces oxidation of phospholipids to initiate ferroptosis.Competing Interest StatementThe authors have declared no competing interest.