Arginine 65 Methylation of Neurogenin 3 by PRMT1 Is Required for Pancreatic Endocrine Development of hESCs

Abstract

Neurogenin3 (NGN3) is a key transcription factor in the cell fate determination of endocrine progenitors (EPs) in the developing pancreas. Previous studies have shown that the stability and activity of NGN3 are regulated by phosphorylation. However, the role of NGN3 methylation is poorly understood. Here, we report that protein arginine methyltransferase-1 (PRMT1)-mediated arginine 65 methylation of NGN3 is required for the pancreatic endocrine development of human embryonic stem cells (hESCs) in vitro. We found that inducible PRMT1-knockout (P-iKO) hESCs did not differentiate from EPs to endocrine cells (ECs) in the presence of doxycycline. Loss of PRMT1 caused NGN3 accumulation in the cytoplasm of EPs and decreased the transcriptional activity of NGN3. We investigated that PRMT1 specifically methylates NGN3 arginine 65, this modification is a prerequisite for ubiquitin-mediated degradation. Our findings demonstrate that arginine 65 methylation of NGN3 is a key molecular switch in hESCs, permitting the differentiation into pancreatic ECs.

Funding Information: This work was supported by Korean Fund for Regenerative Medicine (KFRM) 21A0402L1-12, and the National Research Foundation (NRF) 2021R1A2C300510811, Republic of Korea.

Declaration of Interests: The authors declare no competing interests.

Ethics Approval Statement: Korea advanced institute of science and technology IRB approved the usage of hESCs conducted in this manuscript (KH2021-191).

Keywords: Arginine methylation, NGN3, PRMT1, Pancreatic endocrine cell development, hESC