Abstract
Cell differentiation involves widespread epigenetic reprogramming, including modulation of DNA methylation patterns. The differentiation potential differences in DNA methylation patterns might function in pluripotency restriction, while tissue-specific differences might work in lineage restriction. To investigate the effects of neuronal induction on promoter methylation pattern in rat bone marrow mesenchymal stem cells (MSCs), we used bisulfite sequencing to analyze the methylation status of the promoter regions in neuron-specific enolase (NSE), microtubule-associated protein Tau, and Oct4 genes in MSCs pre- and post-chemical induction. Neurocytes from the newborn rat brains were used as control. Data showed that NSE and Tau were abundantly expressed in the brain cells and MSC-derived neurocyte-like cells as well but not in the MSCs. However, both NSE promoter (−214 ∼ +57 bp) and Tau promoter (−239 ∼ +131 bp) were hypomethylated (<4 % CpG methylation). Oct4 was expressed in MSCs, and the Oct4 promoter (−293 ∼ −85 bp) was hypermethylated (>79 % CpG methylation). Interestingly, it was found that the methylation of the locus −113 bp upstream of Oct4 transcription start site was specifically enhanced in the process of MSCs' neuronal differentiation. Further experiments in hepatocytes derived from MSCs and hepar tissue proved that the −113 bp locus methylation increased also in non-neurogenic lineages. Tfsitescan prediction showed that AP-2-alpha/gamma and Sp1 might regulate Oct4 transcription upon MSC differentiation by binding the −113 bp locus. So, we conclude that promoter methylation modifies pluripotency-specific gene, rather than regulates the expression of neural-specific genes when MSCs differentiate into neurocyte-like cells.
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References
Ballestar E. An introduction to epigenetics. Cell Biology of Stem Cells 711: 1–11; 2011.
Billon N.; Carlisi D.; Datto M. B.; Van Grunsven L.; Watt A.; Wang X. F.; Rudkin B. B. Cooperation of Sp1 and p300 in the induction of the CDK inhibitor p21WAF1/CIP1 during NGF-mediated neuronal differentiation. Oncogene 18: 2872; 1999.
Bird A. P.; Wolffe A. P. Methylation-induced repression--belts, braces, and chromatin. Cell 99: 451; 1999.
Eden S.; Hashimshony T.; Keshet I.; Cedar H.; Thorne A. DNA methylation models histone acetylation. Nature 394: 842–842; 1998.
Fan Z.; Yamaza T.; Lee J. S.; Yu J.; Wang S.; Fan G.; Shi S.; Wang C. Y. BCOR regulates mesenchymal stem cell function by epigenetic mechanisms. Nature Cell Biology 11: 1002–1009; 2009.
Farthing C. R.; Ficz G.; Ng R. K.; Chan C. F.; Andrews S.; Dean W.; Hemberger M.; Reik W. Global mapping of DNA methylation in mouse promoters reveals epigenetic reprogramming of pluripotency genes. PLoS Genet 4: e1000116; 2008.
Gardiner-Garden M.; Frommer M. CpG islands in vertebrate genomes. J. Mol. Biol. 196: 261–282; 1987.
Greco S. J.; Liu K.; Rameshwar P. Functional similarities among genes regulated by OCT4 in human mesenchymal and embryonic stem cells. Stem Cells 25: 3143–3154; 2007.
Gruenbaum Y.; Stein R.; Cedar H.; Razin A. Methylation of CpG sequences in eukaryotic DNA. Febs Letters 124: 67; 1981.
Hilger-Eversheim K.; Moser M.; Schorle H.; Buettner R. Regulatory roles of AP-2 transcription factors in vertebrate development, apoptosis and cell-cycle control. Gene 260: 1–12; 2000.
Imamura T.; Ohgane J.; Ito S.; Ogawa T.; Hattori N.; Tanaka S.; Shiota K. CpG island of rat sphingosine kinase-1 gene: tissue-dependent DNA methylation status and multiple alternative first exons. Genomics 76: 117–125; 2001.
Irizarry R. A; Ladd-Acosta C.; Wen B.; Wu Z.; Montano C.; Onyango P.; Cui H.; Gabo K.; Rongione M.; Webster M. The human colon cancer methylome shows similar hypo-and hypermethylation at conserved tissue-specific CpG island shores. Nature genetics 41: 178–186; 2009.
Jones P. A.; Takai D. The role of DNA methylation in mammalian epigenetics. Science 293: 1068; 2001.
Kadonaga J. T.; Carner K. R.; Masiarz F. R.; Tjian R. Isolation of cDNA encoding transcription factor Sp1 and functional analysis of the DNA binding domain. Cell 51: 1079–1090; 1987.
Li P.; Tong C.; Mehrian-Shai R.; Jia L.; Wu N.; Yan Y.; Maxson R.; Schulze E.; Song H.; Hsieh C. Germline competent embryonic stem cells derived from rat blastocysts. Cell 135: 1299–1310; 2008.
Li Z.; Liu C.; Xie Z.; Song P.; Zhao R. C.; Guo L.; Liu Z.; Wu Y. Epigenetic dysregulation in mesenchymal stem cell aging and spontaneous differentiation. PLoS One 6: e20526; 2011.
Meissner A.; Mikkelsen T. S.; Gu H.; Wernig M.; Hanna J.; Sivachenko A.; Zhang X.; Bernstein B. E.; Nusbaum C.; Jaffe D. B.; Gnirke A.; Jaenisch R.; Lander E. S. Genome-scale DNA methylation maps of pluripotent and differentiated cells. Nature 454: 766–770; 2008.
Mohibullah N.; Donner A.; Ippolito J. A.; Williams T. SELEX and missing phosphate contact analyses reveal flexibility within the AP-2[alpha] protein: DNA binding complex. Nucleic Acids Research 27: 2760–2769; 1999.
Mohn F.; Weber M.; Rebhan M.; Roloff T. C.; Richter J.; Stadler M. B.; Bibel M.; Schubeler D. Lineage-specific polycomb targets and de novo DNA methylation define restriction and potential of neuronal progenitors. Mol Cell 30: 755–766; 2008.
Pittenger M. F.; Mackay A. M.; Beck S. C.; Jaiswal R. K.; Douglas R.; Mosca J. D.; Moorman M. A.; Simonetti D. W.; Craig S.; Marshak D. R. Multilineage potential of adult human mesenchymal stem cells. Science 284: 143–147; 1999.
Reik W.; Dean W.; Walter J. Epigenetic reprogramming in mammalian development. Science 293: 1089; 2001.
Reyes M.; Lund T.; Lenvik T.; Aguiar D.; Koodie L.; Verfaillie C. M. Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood 98: 2615–2625; 2001.
Sensebe L.; Krampera M.; Schrezenmeier H.; Bourin P.; Giordano R. Mesenchymal stem cells for clinical application. Vox Sanguinis 98: 93–107; 2010.
Song F.; Mahmood S.; Ghosh S.; Liang P.; Smiraglia D. J.; Nagase H.; Held W. A. Tissue specific differentially methylated regions (TDMR): Changes in DNA methylation during development. Genomics 93: 130–139; 2009.
Weber M.; Hellmann I.; Stadler M. B.; Ramos L.; Paabo S.; Rebhan M.; Schubeler D. Distribution, silencing potential and evolutionary impact of promoter DNA methylation in the human genome. Nat Genet 39: 457–466; 2007.
Wen-Hai Y.; Xuan-Hui X.; Xue-Fei H.; Lan M.; Yan X.; Jian-Zhi W.; Ying X. Expression of Tau protein and neuron-specific enolase after differentiation of mesenchymal stem cells into neural cells induced by EGF and bFGF. Chinese Journal of Anatomy 6: 615–617; 2005.
Wenke A. K.; Grassel S.; Moser M.; Bosserhoff A. K. The cartilage-specific transcription factor Sox9 regulates AP-2epsilon expression in chondrocytes. Febs Journal 276: 2494–2504; 2009.
Yagi S.; Hirabayashi K.; Sato S.; Li W.; Takahashi Y.; Hirakawa T.; Wu G.; Hattori N.; Ohgane J.; Tanaka S.; Liu X. S.; Shiota K. DNA methylation profile of tissue-dependent and differentially methylated regions (T-DMRs) in mouse promoter regions demonstrating tissue-specific gene expression. Genome Res 18: 1969–1978; 2008.
Yan W.; Cao M.; Liu J.; Xu Y.; Han X.; Xing Y.; Wang J. Effects of EGF and bFGF on expression of microtubule-associated protein tau and MAP-2 mRNA in human umbilical cord mononuclear cells. Cell biology international 29: 153–157; 2005.
Acknowledgments
This study was supported partly by the National Natural Science Foundation of China (no. 30940025) and Science Foundation of Henan Province (no. 0611043000). We thank our colleague Liu Yongmin for the care of the rats.
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Duan, P., Zhang, Y., Han, X. et al. Effect of neuronal induction on NSE, Tau, and Oct4 promoter methylation in bone marrow mesenchymal stem cells. In Vitro Cell.Dev.Biol.-Animal 48, 251–258 (2012). https://doi.org/10.1007/s11626-012-9494-z
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DOI: https://doi.org/10.1007/s11626-012-9494-z