Ubiquitination-deficit of Cnot4 impairs the capacity of proliferation and differentiation in mouse embryonic stem cells

Wenxin Ding; Chenyao He; Xin Liu; Chunlei Hou; Qi Wang; Tiantian Gong; Jiahao Yang; Jingling Shen; Zhiyan Shan; Ruizhen Sun

doi:10.1016/j.bbrc.2024.151260

Ubiquitination-deficit of Cnot4 impairs the capacity of proliferation and differentiation in mouse embryonic stem cells

Biochem Biophys Res Commun. 2024 Dec 27:747:151260. doi: 10.1016/j.bbrc.2024.151260. Online ahead of print.

Authors

Wenxin Ding¹, Chenyao He¹, Xin Liu¹, Chunlei Hou¹, Qi Wang¹, Tiantian Gong¹, Jiahao Yang¹, Jingling Shen², Zhiyan Shan³, Ruizhen Sun⁴

Affiliations

¹ Department of Histology and Embryology, School of Basic Medical Sciences, Harbin Medical University, Harbin, 150081, China.
² Institute of Life Sciences, College of Life and Environmental Science, Wenzhou University, Wenzhou, 325035, China. Electronic address: jingling_shen@wzu.edu.cn.
³ Department of Histology and Embryology, School of Basic Medical Sciences, Harbin Medical University, Harbin, 150081, China. Electronic address: szy@hrbmu.edu.cn.
⁴ Department of Histology and Embryology, School of Basic Medical Sciences, Harbin Medical University, Harbin, 150081, China. Electronic address: srz_1983@hrbmu.edu.cn.

PMID: 39798536
DOI: 10.1016/j.bbrc.2024.151260

Abstract

Neurodevelopmental abnormalities are significant contributors to a variety of neurological disorders. Ubiquitination is essential for embryonic development and plays a pivotal role in neurodevelopment. Although Cnot4 possesses E3-ubiquitin ligase activity, its function in neurodevelopment and embryonic stem cells (ESCs) remains inadequately understood. This study examined the impact of Cnot4 ubiquitination-deficit in mouse ESCs using flow cytometry, CCK-8 assays, immunofluorescence, western blotting, RNA sequencing (RNA-seq), and intracellular Ca²⁺ measurement. Findings demonstrated that the lack of ubiquitination in Cnot4 reduced ESC proliferation rates and facilitated ectodermal differentiation during spontaneous ESC differentiation. RNA-seq analysis identified that the differentially expressed genes were primarily linked to glucose metabolism and Ca²⁺ signaling pathways. Additionally, results indicated that the ubiquitination-deficit in Cnot4 caused increased intracellular Ca²⁺ levels in mESCs. These findings suggest that Cnot4 plays a critical role in the regulation of proliferation and differentiation of mESCs through ubiquitination, providing a basis for further exploration of its involvement in embryonic and neural development.

Keywords: Cnot4; Embryonic stem cells; Neural differentiation; Ubiquitination.