Abstract 7081: Comprehensive investigation of NAT10’s catalytic and non-catalytic functions in cancer cell proliferation

Abstract

Abstract N-acetyltransferase 10 (NAT10) is emerging as an oncoprotein in multiple cancers including acute myeloid leukemia and liver cancer. In addition, my preliminary studies show that NAT10 is a stress response that promotes the resilience of cancer cells to anti-tumor drugs. However, the underlying mechanisms by which NAT10 promotes cancer growth and drug resistance are unknown. NAT10 is an RNA acetyltransferase, where it acetylates cytidines in rRNA, tRNA, mRNA, and lncRNAs. Our previous work demonstrated that NAT10 promotes translation efficiency and mRNA stability through the acetylation of cytidines in mRNA. However, NAT10 is a multifunctional protein. Besides acetylating RNA, NAT10 also acetylates proteins at lysine residues, has RNA helicase activity, and can serve as a scaffold in ribonucleoprotein complexes. Based on this complexity, it is imperative to elucidate whether NAT10 exerts its oncogenic activity via RNA acetylation, protein acetylation, disruption of RNA structures, or non-catalytic functions. Strikingly, our preliminary results indicate that the acetyltransferase function of NAT10 is dispensable for cell proliferation. To evaluate NAT10’s oncogenic activity unbiasedly, we are now performing a deep mutagenesis scanning (DMS) assay. For this purpose, we generated a library of mutants in hundreds of NAT10 residues. This library was transduced in multiple cell lines from acute myeloid leukemia and liver cancer carrying a PROTAC system, which allow us to deplete NAT10 in a rapid and controlled manner while expressing ectopically thousands of NAT10 mutants. In this conference, I will talk about, my results, how DMS is a powerful tool to delineate the molecular functions of candidate oncoproteins, and the implications of our results for the potential drugging of NAT10 in cancer. Keywords: Cell proliferation, NAT10, RNA modifications, Deep mutagenesis scanning. Citation Format: Sweta Pradeep Raikundalia, Mahmood Hassan Dalhat, Emmely A. Patrasso, Adam Suh, Daniel Arango. Comprehensive investigation of NAT10’s catalytic and non-catalytic functions in cancer cell proliferation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7081.