title: Autophagy regulates homeostasis of pluripotency-associated proteins in hESCs.
abstract: The pluripotency of embryonic stem cells (ESCs) is maintained by intracellular networks of many pluripotency-associated (PA) proteins such as OCT4, SOX2, and NANOG. However, the mechanisms underlying the regulation of protein homeostasis for pluripotency remain elusive. Here, we first demonstrate that autophagy acts together with the ubiquitin-proteasome system (UPS) to modulate the levels of PA proteins in human ESCs (hESCs). Autophagy inhibition impaired the pluripotency despite increment of PA proteins in hESCs. Immunogold-electron microscopy confirmed localization of OCT4 molecules within autophagosomes. Also, knockdown of LC3 expression led to accumulation of PA proteins and reduction of pluripotency in hESCs. Interestingly, autophagy and the UPS showed differential kinetics in the degradation of PA proteins. Autophagy inhibition caused enhanced accumulation of both cytoplasmic and nuclear PA proteins, whereas the UPS inhibition led to preferentially degrade nuclear PA proteins. Our findings suggest that autophagy modulates homeostasis of PA proteins, providing a new insight in the regulation of pluripotency in hESCs.
authors.affiliation: Department of Biological Sciences and Center for Stem Cell Differentiation, Daejeon, Republic of Korea.
authors.forename: Yun-Hee
authors.initials: YH
authors.lastname: Cho
authors.forename: Kyu-Min
authors.initials: KM
authors.lastname: Han
authors.forename: Dongkyu
authors.initials: D
authors.lastname: Kim
authors.forename: Joonsun
authors.initials: J
authors.lastname: Lee
authors.forename: Sang-Hee
authors.initials: SH
authors.lastname: Lee
authors.forename: Kyeng-Won
authors.initials: KW
authors.lastname: Choi
authors.forename: Jungho
authors.initials: J
authors.lastname: Kim
authors.forename: Yong-Mahn
authors.initials: YM
authors.lastname: Han
mesh_terms: D001343:Autophagy / Q000235:genetics*; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000378:metabolism; D006706:Homeostasis; D006801:Humans; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D025801:Ubiquitin / Q000235:genetics
chemical_list: D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D025801:Ubiquitin; D046988:Proteasome Endopeptidase Complex
keywords: Cell culture; Cell signaling; Embryonic stem cells; Pluripotency