OCT4 translationally promotes AKT signaling as an RNA-binding protein in stressed pluripotent stem cells
Abstract Background Despite numerous studies addressing the molecular mechanisms by which pluripotent stem cells (PSCs) maintain self-renewal and pluripotency under normal culture conditions, the fundamental question of how PSCs manage to survive stressful conditions remains largely unresolved. Post...
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BMC
2025-02-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | https://doi.org/10.1186/s13287-025-04229-1 |
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| author | Wenjie Chen Xinyu Chen Cheng Chen Shiqi She Xia Li Lina Shan Xiaobing Zhang Songsong Dan Yisha Wang Yan-Wen Zhou Qingyi Cao Wenxin Wang Jianwen Hu Yaxun Wei Yaqiang Xue Yi Zhang Songying Zhang Ying-Jie Wang Bo Kang |
| author_facet | Wenjie Chen Xinyu Chen Cheng Chen Shiqi She Xia Li Lina Shan Xiaobing Zhang Songsong Dan Yisha Wang Yan-Wen Zhou Qingyi Cao Wenxin Wang Jianwen Hu Yaxun Wei Yaqiang Xue Yi Zhang Songying Zhang Ying-Jie Wang Bo Kang |
| author_sort | Wenjie Chen |
| collection | DOAJ |
| description | Abstract Background Despite numerous studies addressing the molecular mechanisms by which pluripotent stem cells (PSCs) maintain self-renewal and pluripotency under normal culture conditions, the fundamental question of how PSCs manage to survive stressful conditions remains largely unresolved. Post-transcriptional/translational regulation emerges to be vital for PSCs, but how PSCs coordinate and balance their survival and differentiation at translational level under extrinsic and intrinsic stress conditions is unclear. Methods The high-throughput sequencing of cross-linking immunoprecipitation cDNA library (HITS-CLIP) was employed to decipher the genome-wide OCT4-RNA interactome in human PSCs, a combined RNC-seq/RNA-seq analysis to assess the role of OCT4 in translational regulation of hypoxic PSCs, and an OCT4-protein interactome to search for OCT4 binding partners that regulate cap-independent translation initiation. By taking the Heterozygous Knocking In N-terminal Tags (HKINT) approach that specifically disrupts the 5′-UTR secondary structure and tagging its protein product of the mRNA from one allele while leaving that from the other allele intact, we examined the effect of disrupting the OCT4/5′-UTR interaction on translation of AKT1 mRNA. Results We revealed OCT4 as a bona fide RNA-binding protein (RBP) in human PSCs that bound to the 5′-UTR, 3′-UTR and CDS regions of mRNAs. Multiple known proteins participating in IRES-mediated translation initiation were detected in the OCT4-protein interactome, and a combined RNC-seq/RNA-seq analysis further confirmed a crucial role of OCT4 in translational regulation of PSCs in response to hypoxic stress. Remarkably, OCT4 bound to the GC-rich elements in the 5′-UTR of AKT1 and multiple PI3K/AKT-pathway-gene mRNAs, and promoted their translation initiation via IRES-mediated pathways under stress conditions. Specifically disrupting the AKT1 mRNA 5′-UTR structure and the OCT4/5′-UTR interaction by the HKINT approach significantly reduced the translation level of AKT1 that led to a higher susceptibility of PSCs to oxidative stress-induced apoptotic death and prioritized differentiation toward ectoderm and endoderm. Conclusions Our results reveal OCT4 as an anti-stress RBP for translational regulation that critically coordinates the survival and differentiation of PSCs in response to various stressors. |
| format | Article |
| id | doaj-art-1b29b56cf5a4439e9075f1e218e0efc3 |
| institution | OA Journals |
| issn | 1757-6512 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | BMC |
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| series | Stem Cell Research & Therapy |
| spelling | doaj-art-1b29b56cf5a4439e9075f1e218e0efc32025-08-20T02:16:34ZengBMCStem Cell Research & Therapy1757-65122025-02-0116112610.1186/s13287-025-04229-1OCT4 translationally promotes AKT signaling as an RNA-binding protein in stressed pluripotent stem cellsWenjie Chen0Xinyu Chen1Cheng Chen2Shiqi She3Xia Li4Lina Shan5Xiaobing Zhang6Songsong Dan7Yisha Wang8Yan-Wen Zhou9Qingyi Cao10Wenxin Wang11Jianwen Hu12Yaxun Wei13Yaqiang Xue14Yi Zhang15Songying Zhang16Ying-Jie Wang17Bo Kang18State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityDepartment of Obstetrics and Gynecology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang UniversityDepartment of Colorectal SurgerySir Run Run Shaw Hospital,, School of Medicine, Zhejiang UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityDepartment of Infectious Diseases, the Second Xiangya Hospital, Central South UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversitySchool of Medicine, Zhejiang UniversityShanghai Bioprofile Technology Co., Ltd.Center for Genome Analysis, ABLife Inc.Center for Genome Analysis, ABLife Inc.Center for Genome Analysis, ABLife Inc.Department of Obstetrics and Gynecology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityAbstract Background Despite numerous studies addressing the molecular mechanisms by which pluripotent stem cells (PSCs) maintain self-renewal and pluripotency under normal culture conditions, the fundamental question of how PSCs manage to survive stressful conditions remains largely unresolved. Post-transcriptional/translational regulation emerges to be vital for PSCs, but how PSCs coordinate and balance their survival and differentiation at translational level under extrinsic and intrinsic stress conditions is unclear. Methods The high-throughput sequencing of cross-linking immunoprecipitation cDNA library (HITS-CLIP) was employed to decipher the genome-wide OCT4-RNA interactome in human PSCs, a combined RNC-seq/RNA-seq analysis to assess the role of OCT4 in translational regulation of hypoxic PSCs, and an OCT4-protein interactome to search for OCT4 binding partners that regulate cap-independent translation initiation. By taking the Heterozygous Knocking In N-terminal Tags (HKINT) approach that specifically disrupts the 5′-UTR secondary structure and tagging its protein product of the mRNA from one allele while leaving that from the other allele intact, we examined the effect of disrupting the OCT4/5′-UTR interaction on translation of AKT1 mRNA. Results We revealed OCT4 as a bona fide RNA-binding protein (RBP) in human PSCs that bound to the 5′-UTR, 3′-UTR and CDS regions of mRNAs. Multiple known proteins participating in IRES-mediated translation initiation were detected in the OCT4-protein interactome, and a combined RNC-seq/RNA-seq analysis further confirmed a crucial role of OCT4 in translational regulation of PSCs in response to hypoxic stress. Remarkably, OCT4 bound to the GC-rich elements in the 5′-UTR of AKT1 and multiple PI3K/AKT-pathway-gene mRNAs, and promoted their translation initiation via IRES-mediated pathways under stress conditions. Specifically disrupting the AKT1 mRNA 5′-UTR structure and the OCT4/5′-UTR interaction by the HKINT approach significantly reduced the translation level of AKT1 that led to a higher susceptibility of PSCs to oxidative stress-induced apoptotic death and prioritized differentiation toward ectoderm and endoderm. Conclusions Our results reveal OCT4 as an anti-stress RBP for translational regulation that critically coordinates the survival and differentiation of PSCs in response to various stressors.https://doi.org/10.1186/s13287-025-04229-1OCT4AKTRNA-binding protein (RBP)Translation initiationOxidative stressPluripotent stem cells (PSCs) |
| spellingShingle | Wenjie Chen Xinyu Chen Cheng Chen Shiqi She Xia Li Lina Shan Xiaobing Zhang Songsong Dan Yisha Wang Yan-Wen Zhou Qingyi Cao Wenxin Wang Jianwen Hu Yaxun Wei Yaqiang Xue Yi Zhang Songying Zhang Ying-Jie Wang Bo Kang OCT4 translationally promotes AKT signaling as an RNA-binding protein in stressed pluripotent stem cells Stem Cell Research & Therapy OCT4 AKT RNA-binding protein (RBP) Translation initiation Oxidative stress Pluripotent stem cells (PSCs) |
| title | OCT4 translationally promotes AKT signaling as an RNA-binding protein in stressed pluripotent stem cells |
| title_full | OCT4 translationally promotes AKT signaling as an RNA-binding protein in stressed pluripotent stem cells |
| title_fullStr | OCT4 translationally promotes AKT signaling as an RNA-binding protein in stressed pluripotent stem cells |
| title_full_unstemmed | OCT4 translationally promotes AKT signaling as an RNA-binding protein in stressed pluripotent stem cells |
| title_short | OCT4 translationally promotes AKT signaling as an RNA-binding protein in stressed pluripotent stem cells |
| title_sort | oct4 translationally promotes akt signaling as an rna binding protein in stressed pluripotent stem cells |
| topic | OCT4 AKT RNA-binding protein (RBP) Translation initiation Oxidative stress Pluripotent stem cells (PSCs) |
| url | https://doi.org/10.1186/s13287-025-04229-1 |
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