Stub1 maintains proteostasis of master transcription factors in embryonic stem cells

Md Mahfuz Al Mamun; Muhammad Riaz Khan; Yifu Zhu; Yuwei Zhang; Shuai Zhou; Ran Xu; Ihtisham Bukhari; Rick F Thorne; Jinming Li; Xu Dong Zhang; Guangzhi Liu; Song Chen; Mian Wu; Xiaoyuan Song

doi:10.1016/j.celrep.2022.110919

Stub1 maintains proteostasis of master transcription factors in embryonic stem cells

Cell Rep. 2022 Jun 7;39(10):110919. doi: 10.1016/j.celrep.2022.110919.

Authors

Md Mahfuz Al Mamun¹, Muhammad Riaz Khan², Yifu Zhu³, Yuwei Zhang⁴, Shuai Zhou¹, Ran Xu⁵, Ihtisham Bukhari¹, Rick F Thorne⁶, Jinming Li¹, Xu Dong Zhang⁷, Guangzhi Liu⁸, Song Chen⁹, Mian Wu¹⁰, Xiaoyuan Song¹¹

Affiliations

¹ Translational Research Institute, Henan Provincial People's Hospital, Henan Key Laboratory of Stem Cell Differentiation and Modification, School of Clinical Medicine, Henan University, Zhengzhou 450003, China; Zhengzhou City Key Laboratory of Long Non-coding RNA and Cancer Metabolism, Henan Provincial Key Laboratory of Long Non-coding RNA and Cancer Metabolism, Henan International Joint Laboratory of Non-coding RNA and Metabolism in Cancer, Zhengzhou 450003, China.
² Translational Research Institute, Henan Provincial People's Hospital, Henan Key Laboratory of Stem Cell Differentiation and Modification, School of Clinical Medicine, Henan University, Zhengzhou 450003, China; Zhengzhou City Key Laboratory of Long Non-coding RNA and Cancer Metabolism, Henan Provincial Key Laboratory of Long Non-coding RNA and Cancer Metabolism, Henan International Joint Laboratory of Non-coding RNA and Metabolism in Cancer, Zhengzhou 450003, China; Research Center on Aging, Centre Intégré Universitaire de Santé et Services Sociaux de l'Estrie-Centre Hospitalier Universitaire de Sherbrooke, Sherbrooke, QC, Canada; Department of Biochemistry and Functional Genomics, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, QC J1E 4K8 Canada.
³ CAS Key Laboratory of Innate Immunity and Chronic Disease, CAS Centre for Excellence in Molecular Cell Science, The First Affiliated Hospital of University of Science and Technology of China, Hefei 230027, China.
⁴ Translational Research Institute, Henan Provincial People's Hospital, Henan Key Laboratory of Stem Cell Differentiation and Modification, School of Clinical Medicine, Henan University, Zhengzhou 450003, China.
⁵ School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, NSW 2308, Australia.
⁶ Translational Research Institute, Henan Provincial People's Hospital, Henan Key Laboratory of Stem Cell Differentiation and Modification, School of Clinical Medicine, Henan University, Zhengzhou 450003, China; Zhengzhou City Key Laboratory of Long Non-coding RNA and Cancer Metabolism, Henan Provincial Key Laboratory of Long Non-coding RNA and Cancer Metabolism, Henan International Joint Laboratory of Non-coding RNA and Metabolism in Cancer, Zhengzhou 450003, China; Molecular Pathology Center, Academy of Medical Science, Zhengzhou University, Zhengzhou 450053, China; School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW 2258, Australia.
⁷ Translational Research Institute, Henan Provincial People's Hospital, Henan Key Laboratory of Stem Cell Differentiation and Modification, School of Clinical Medicine, Henan University, Zhengzhou 450003, China; Zhengzhou City Key Laboratory of Long Non-coding RNA and Cancer Metabolism, Henan Provincial Key Laboratory of Long Non-coding RNA and Cancer Metabolism, Henan International Joint Laboratory of Non-coding RNA and Metabolism in Cancer, Zhengzhou 450003, China; Molecular Pathology Center, Academy of Medical Science, Zhengzhou University, Zhengzhou 450053, China; School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, NSW 2308, Australia.
⁸ Translational Research Institute, Henan Provincial People's Hospital, Henan Key Laboratory of Stem Cell Differentiation and Modification, School of Clinical Medicine, Henan University, Zhengzhou 450003, China. Electronic address: guangzhi72@126.com.
⁹ Translational Research Institute, Henan Provincial People's Hospital, Henan Key Laboratory of Stem Cell Differentiation and Modification, School of Clinical Medicine, Henan University, Zhengzhou 450003, China; Zhengzhou City Key Laboratory of Long Non-coding RNA and Cancer Metabolism, Henan Provincial Key Laboratory of Long Non-coding RNA and Cancer Metabolism, Henan International Joint Laboratory of Non-coding RNA and Metabolism in Cancer, Zhengzhou 450003, China; Molecular Pathology Center, Academy of Medical Science, Zhengzhou University, Zhengzhou 450053, China; Institute of Medicinal Biotechnology, Jiangsu College of Nursing, Huai'an, Jiangsu 223300, China. Electronic address: schen@zzu.edu.cn.
¹⁰ Translational Research Institute, Henan Provincial People's Hospital, Henan Key Laboratory of Stem Cell Differentiation and Modification, School of Clinical Medicine, Henan University, Zhengzhou 450003, China; Zhengzhou City Key Laboratory of Long Non-coding RNA and Cancer Metabolism, Henan Provincial Key Laboratory of Long Non-coding RNA and Cancer Metabolism, Henan International Joint Laboratory of Non-coding RNA and Metabolism in Cancer, Zhengzhou 450003, China; CAS Key Laboratory of Innate Immunity and Chronic Disease, CAS Centre for Excellence in Molecular Cell Science, The First Affiliated Hospital of University of Science and Technology of China, Hefei 230027, China; Molecular Pathology Center, Academy of Medical Science, Zhengzhou University, Zhengzhou 450053, China. Electronic address: wumian@ustc.edu.cn.
¹¹ MOE Key Laboratory for Cellular Dynamics, Hefei National Research Center for Physical Sciences at the Microscale, CAS Key Laboratory of Brain Function and Disease, School of Life Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China. Electronic address: songxy5@ustc.edu.cn.

PMID: 35675767
DOI: 10.1016/j.celrep.2022.110919

Abstract

The pluripotency and differentiation states of embryonic stem cells (ESCs) are regulated by a set of core transcription factors, primarily Sox2, Oct4, and Nanog. Although their transcriptional regulation has been studied extensively, the contribution of posttranslational modifications in Sox2, Oct4, and Nanog are poorly understood. Here, using a CRISPR-Cas9 knockout library screen in murine ESCs, we identify the E3 ubiquitin ligase Stub1 as a negative regulator of pluripotency. Manipulation of Stub1 expression in murine ESCs shows that ectopic Stub1 expression significantly reduces the protein half-life of Sox2, Oct4, and Nanog. Mechanistic investigations reveal Stub1 catalyzes the polyubiquitination and 26S proteasomal degradation of Sox2 and Nanog through K48-linked ubiquitin chains and Oct4 via K63 linkage. Stub1 deficiency positively enhances somatic cell reprogramming and delays differentiation, whereas its enforced expression triggers ESC differentiation. The discovery of Stub1 as an integral pluripotency regulator strengthens our understanding of ESC regulation beyond conventional transcriptional control mechanisms.

Keywords: CP: Molecular biology; CP: Stem cell research; Nanog; Oct4; Sox2; Stub1; differentiation; embryonic stem cells; induced pluripotent stem cells; pluripotency; ubiquitination.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Differentiation / physiology
Embryonic Stem Cells* / metabolism
Gene Expression Regulation
Homeodomain Proteins / metabolism
Mice
Nanog Homeobox Protein / metabolism
Octamer Transcription Factor-3 / metabolism
Proteostasis*
SOXB1 Transcription Factors / metabolism
Transcription Factors* / metabolism
Ubiquitin-Protein Ligases* / metabolism

Substances

Homeodomain Proteins
Nanog Homeobox Protein
Octamer Transcription Factor-3
SOXB1 Transcription Factors
Transcription Factors
Stub1 protein, mouse
Ubiquitin-Protein Ligases