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O-GlcNAcylation of Sox2 at threonine 258 regulates the self-renewal and early cell fate of embryonic stem cells

김희연|최지웅|신현무|윤홍덕|Jang Hyonchol|Lee Eun Young|Kim Dong Keon|Lee Jang-Seok|Jang Hansol|Han Suji|Hwang In-Young|You Hye Jin
생화학분자생물학회 (Korean Society Of Biochemistry And Molecular Biology)|2021.12|Experimental and Molecular Medicine|제53권 |pp. 1-10|DOI 10.1038/s12276-021-00707-7

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주제분류생화학

초록

Sox2 is a core transcription factor in embryonic stem cells (ESCs), and O -GlcNAcylation is a type of post-translational modification of nuclear-cytoplasmic proteins. Although both factors play important roles in the maintenance and differentiation of ESCs and the serine 248 (S248) and threonine 258 (T258) residues of Sox2 are modified by O -GlcNAcylation, the function of Sox2 O -GlcNAcylation is unclear. Here, we show that O -GlcNAcylation of Sox2 at T258 regulates mouse ESC self-renewal and early cell fate. ESCs in which wild-type Sox2 was replaced with the Sox2 T258A mutant exhibited reduced self-renewal, whereas ESCs with the Sox2 S248A point mutation did not. ESCs with the Sox2 T258A mutation heterologously introduced using the CRISPR/Cas9 system, designated E14-Sox2 TA/WT , also exhibited reduced self-renewal. RNA sequencing analysis under self-renewal conditions showed that upregulated expression of early differentiation genes, rather than a downregulated expression of self-renewal genes, was responsible for the reduced self-renewal of E14-Sox2 TA/WT cells. There was a significant decrease in ectodermal tissue and a marked increase in cartilage tissue in E14-Sox2 TA/WT -derived teratomas compared with normal E14 ESC-derived teratomas. RNA sequencing of teratomas revealed that genes related to brain development had generally downregulated expression in the E14-Sox2 TA/WT -derived teratomas. Our findings using the Sox2 T258A mutant suggest that Sox2 T258 O -GlcNAc has a positive effect on ESC self-renewal and plays an important role in the proper development of ectodermal lineage cells. Overall, our study directly links O -GlcNAcylation and early cell fate decisions.

Abstract

Sox2 is a core transcription factor in embryonic stem cells (ESCs), and O -GlcNAcylation is a type of post-translational modification of nuclear-cytoplasmic proteins. Although both factors play important roles in the maintenance and differentiation of ESCs and the serine 248 (S248) and threonine 258 (T258) residues of Sox2 are modified by O -GlcNAcylation, the function of Sox2 O -GlcNAcylation is unclear. Here, we show that O -GlcNAcylation of Sox2 at T258 regulates mouse ESC self-renewal and early cell fate. ESCs in which wild-type Sox2 was replaced with the Sox2 T258A mutant exhibited reduced self-renewal, whereas ESCs with the Sox2 S248A point mutation did not. ESCs with the Sox2 T258A mutation heterologously introduced using the CRISPR/Cas9 system, designated E14-Sox2 TA/WT , also exhibited reduced self-renewal. RNA sequencing analysis under self-renewal conditions showed that upregulated expression of early differentiation genes, rather than a downregulated expression of self-renewal genes, was responsible for the reduced self-renewal of E14-Sox2 TA/WT cells. There was a significant decrease in ectodermal tissue and a marked increase in cartilage tissue in E14-Sox2 TA/WT -derived teratomas compared with normal E14 ESC-derived teratomas. RNA sequencing of teratomas revealed that genes related to brain development had generally downregulated expression in the E14-Sox2 TA/WT -derived teratomas. Our findings using the Sox2 T258A mutant suggest that Sox2 T258 O -GlcNAc has a positive effect on ESC self-renewal and plays an important role in the proper development of ectodermal lineage cells. Overall, our study directly links O -GlcNAcylation and early cell fate decisions.

참고문헌 (53)

  1. Hardiville, S., "Nutrient regulation of signaling, transcription, and cell physiology by O-GlcNAcylation", Cell Metab, 20, pp.208~213 (2014)
  2. Schjoldager, K. T., "Global view of human protein glycosylation pathways and functions", Nat. Rev. Mol. Cell Biol., 21, pp.729~749 (2020)
  3. Jang, H., "O-GlcNAc regulates pluripotency and reprogramming by directly acting on core components of the pluripotency network", Cell Stem Cell, 11, pp.62~74 (2012)
  4. Zhu, Q., "O-GlcNAcylation regulates the methionine cycle to promote pluripotency of stem cells", Proc. Natl Acad. Sci. USA, 117, pp.7755~7763 (2020)
  5. Hao, Y., "Next-generation unnatural monosaccharides reveal that ESRRB O-GlcNAcylation regulates pluripotency of mouse embryonic stem cells", Nat. Commun, 10, pp.4065~ (2019)
  6. Andres, L. M., "Chemical modulation of protein O-GlcNAcylation via OGT inhibition promotes human neural cell differentiation", ACS Chem. Biol, 12, pp.2030~2039 (2017)
  7. Kim, H. S., "Excessive O-GlcNAcylation of proteins suppresses spontaneous cardiogenesis in ES cells", FEBS Lett, 583, pp.2474~2478 (2009)
  8. Li, M., "Deconstructing the pluripotency gene regulatory network", Nat. Cell Biol, 20, pp.382~392 (2018)
  9. Chambers, I., "The transcriptional foundation of pluripotency", Development, 136, pp.2311~2322 (2009)
  10. Boyer, L. A., "Core transcriptional regulatory circuitry in human embryonic stem cells", Cell, 122, pp.947~956 (2005)
  11. Lee, T. I., "Control of developmental regulators by Polycomb in human embryonic stem cells", Cell, 125, pp.301~313 (2006)
  12. Avilion, A. A., "Multipotent cell lineages in early mouse development depend on SOX2 function", Genes Dev, 17, pp.126~140 (2003)
  13. Thomson, M., "Pluripotency factors in embryonic stem cells regulate differentiation into germ layers", Cell, 145, pp.875~889 (2011)
  14. Takemoto, T., "Tbx6-dependent Sox2 regulation determines neural or mesodermal fate in axial stem cells", Nature, 470, pp.394~398 (2011)
  15. Sarkar, A., "The sox family of transcription factors : versatile regulators of stem and progenitor cell fate", Cell Stem Cell, 12, pp.15~30 (2013)
  16. Kamachi, Y., "Sox proteins : regulators of cell fate specification and differentiation", Development, 140, pp.4129~4144 (2013)
  17. Zhang, S., "Functional characterization of SOX2 as an anticancer target", Signal Transduct. Target. Ther, 5, pp.135~ (2020)
  18. Schaefer, T., "SOX2 protein biochemistry in stemness, reprogramming, and cancer : the PI3K", AKT, Oncogene, pp.2020, 39[DOI]
  19. Khidekel, N., "Exploring the O-GlcNAc proteome : direct identification of O-GlcNAc-modified proteins from the brain", Proc. Natl Acad. Sci. USA, 101, pp.13132~13137 (2004)
  20. Myers, S. A., "Polycomb repressive complex 2 is necessary for the normal site-specific O-GlcNAc distribution in mouse embryonic stem cells", Proc. Natl Acad. Sci. USA, 108, pp.9490~9495 (2011)
  21. Myers, S. A., "SOX2 O-GlcNAcylation alters its protein-protein interactions and genomic occupancy to modulate gene expression in pluripotent cells", Elife, 5, pp.e10647~ (2016)
  22. Hwang, I. Y., "Psat1-dependent fluctuations in alpha-ketoglutarate affect the timing of ESC differentiation", Cell Metab, 24, pp.494~501 (2016)
  23. Bong, S. M., "Regulation of mRNA export through API5 and nuclear FGF2interaction", Nucleic Acids Res, 48, pp.6340~6352 (2020)
  24. 김희연, "Farnesyl diphosphate synthase is important for the maintenance of glioblastoma stemness", Experimental and Molecular Medicine, 50, pp.1~12 (2018), 생화학분자생물학회
  25. Kim, D. K., "Phosphorylation of OCT4 serine 236 inhibits germ cell tumor growth by inducing differentiation", Cancers, 12, pp.2601~ (2020)
  26. Kim, H., "Core Pluripotency Factors Directly Regulate Metabolism in Embryonic Stem Cell to Maintain Pluripotency", Stem Cells, 33, pp.2699~2711 (2015)
  27. Abulaiti, X., "Phosphorylation of threonine(343)is crucial for OCT4 interaction with SOX2 in the maintenance of mouse embryonic stem cell pluripotency", Stem Cell Rep, 9, pp.1630~1641 (2017)
  28. Jang, H., "Cabin1 restrains p53 activity on chromatin", Nat. Struct. Mol. Biol, 16, pp.910~915 (2009)
  29. POOJAN SHIV, "Cancer cells undergoing epigenetic transition show short-term resistance and are transformed into cells with medium-term resistance by drug treatment", Experimental and Molecular Medicine, 52, pp.1~14 (2020), 생화학분자생물학회[DOI]
  30. Lee, J. S., "SEZ6L2 is an important regulator of drug-resistant cells and tumor spheroid cells in lung adenocarcinoma", Biomedicines, 8, pp.500~ (2020)
  31. Kwak, S., "Zinc finger proteins orchestrate active gene silencing during embryonic stem cell differentiation", Nucleic Acids Res, 46, pp.6592~6607 (2018)
  32. Whyte, W. A., "Master transcription factors and mediator establish superenhancers at key cell identity genes", Cell, 153, pp.307~319 (2013)
  33. Robinson, J. T., "Integrative genomics viewer", Nat. Biotechnol, 29, pp.24~26 (2011)
  34. Kim, H., "A quenched annexin V-fluorophore for the real-time fluorescence imaging of apoptotic processes in vitro and in vivo", Adv. Sci, 7, pp.2002988~ (2020)
  35. Swaney, D. L., "Human embryonic stem cell phosphoproteome revealed by electron transfer dissociation tandem mass spectrometry", Proc. Natl Acad. Sci. USA, 106, pp.995~1000 (2009)
  36. Rigbolt, K. T., "System-wide temporal characterization of the proteome and phosphoproteome of human embryonic stem cell differentiation", Sci. Signal., 4, pp.rs3~ (2011)
  37. Masui, S., "Pluripotency governed by Sox2 via regulation of Oct3", 4 expression in mouse embryonic stem cells, 2007, pp.9, 625~635
  38. Pannell, D., "Silencing of gene expression : implications for design of retrovirus vectors", Rev. Med. Virol, 11, pp.205~217 (2001)
  39. Niwa, H., "Efficient selection for high-expression transfectants with a novel eukaryotic vector", Gene, 108, pp.193~199 (1991)
  40. Jang, J., "Nrf2, a regulator of the proteasome, controls self-renewal and pluripotency in human embryonic stem cells", Stem Cells, 32, pp.2616~2625 (2014)
  41. Wang, Q. Y., "Glutathione peroxidase-1 is required for self-renewal of murine embryonic stem cells", Biochem. Biophys. Res. Commun, 448, pp.454~460 (2014)
  42. Han, X., "Nonsense-mediated mRNA decay : a ‘nonsense’ pathway makes sense in stem cell biology", Nucleic Acids Res, 46, pp.1038~1051 (2018)
  43. metabolism controls stem cell identity, "Cell Death Differ", 2015, 1094~1105
  44. Lu, R., "Dual functions of T-box 3(Tbx3)in the control of selfrenewal and extraembryonic endoderm differentiation in mouse embryonic stem cells", J. Biol. Chem, 286, pp.8425~8436 (2011)
  45. Savarese, F., "Satb1 and Satb2 regulate embryonic stem cell differentiation and Nanog expression", Genes Dev, 23, pp.2625~2638 (2009)
  46. Yang, Y. R., "O-GlcNAcase is essential for embryonic development and maintenance of genomic stability", Aging Cell, 11, pp.439~448 (2012)
  47. Muha, V., "Loss of O-GlcNAcase catalytic activity leads to defects in mouse embryogenesis", J. Biol. Chem, 296, pp.100439~ (2021)
  48. Shafi, R., "The O-GlcNAc transferase gene resides on the X chromosome and is essential for embryonic stem cell viability and mouse ontogeny", Proc. Natl Acad. Sci. USA, 97, pp.5735~5739 (2000)
  49. Takashima, Y., "Resetting transcription factor control circuitry toward groundstate pluripotency in human", Cell, 158, pp.1254~1269 (2014)
  50. Kopp, J. L., "Small increases in the level of Sox2 trigger the differentiation of mouse embryonic stem cells", Stem Cells, 26, pp.903~911 (2008)
  51. Metz, E. P., "Sox2 dosage : a critical determinant in the functions of Sox2 in both normal and tumor cells", J. Cell. Physiol, 234, pp.19298~19306 (2019)
  52. Shu, J., "Induction of pluripotency in mouse somatic cells with lineage specifiers", Cell, 153, pp.963~975 (2013)
  53. Holmes, G., "The Sox2 high mobility group transcription factor inhibits mature osteoblast function in transgenic mice", Bone, 49, pp.653~661 (2011)

관련 논문AI추천

OCT4와 SOX2에 의한 인간 Nanog 유전자의 전사 조절김영은, 박정아, 석현정, 이영희한국발생생물학회 · 발생과 생식2010 · 제14권 제2호Oct4 resetting by Aurkb–PP1 cell cycle axis determines the identity of mouse embryonic stem cells신지훈, 윤홍덕생화학분자생물학회 · BMB Reports2016 · 제49권 제10호In vivo putative O-GlcNAcylation of human SCP1 and evidence for possible role of its N-terminal disordered structure구재형, 박영일생화학분자생물학회 · BMB Reports2014 · 제47권 제10호Translationally controlled tumor protein (TCTP) downregulates Oct4 expression in mouse pluripotent cellsHuayan Wang, Jie Deng, Junhua Li, Shuyan Meng, Xiang Cheng, Zhenzhen Li생화학분자생물학회 · BMB Reports2012 · 제45권 제1호Specific Knockdown of Nanog Expression by RNA Interference in P19 Embryonal Carcinoma Stem Cells임도선, 최승철한국발생생물학회 · 발생과 생식2008 · 제12권 제2호Suppression of SIRT2 and altered acetylation status of human pluripotent stem cells: possible link to metabolic switch during reprogramming권옥선, 차혁진, 한민준생화학분자생물학회 · BMB Reports2017 · 제50권 제9호

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