TY - JOUR
T1 - Protein arginine methyltransferase 7-mediated microRNA- 221 repression maintains Oct4, Nanog, and Sox2 levels in mouse embryonic stem cells
AU - Chen, Tsai Yu
AU - Lee, Sung Hun
AU - Dhar, Shilpa S.
AU - Lee, Min Gyu
N1 - Funding Information:
This study was supported by National Institutes of Health Grants R01 CA207098, R01 CA207109, and R01 GM095659 (to M.G.L.); the Cancer Pre-vention and Research Institute of Texas Grant RP140271 (to M. G. L.); and the Center for Cancer Epigenetics at M. D. Anderson Grant (to M. G. L.). The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Funding Information:
Acknowledgments—We thank the research staff in the Flow Cytometry & Cellular Imaging Facility, which is supported in part by the National Institutes of Health through M. D. Anderson’s Cancer Center Support Grant CA016672. We also thank Ann Sutton (Department of Scientific Publications, M. D. Anderson) for the editorial assistance.
Funding Information:
This study was supported by National Institutes of Health Grants R01 CA207098, R01 CA207109, and R01 GM095659 (to M.G.L.); the Cancer Prevention and Research Institute of Texas Grant RP140271 (to M. G. L.); and the Center for Cancer Epigenetics at M. D. Anderson Grant (to M. G. L.). The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Publisher Copyright:
© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2018/3/16
Y1 - 2018/3/16
N2 - The stemness maintenance of embryonic stem cells (ESCs) requires pluripotency transcription factors, including Oct4, Nanog, and Sox2. We have previously reported that protein arginine methyltransferase 7 (PRMT7), an epigenetic modifier, is an essential pluripotency factor that maintains the stemness of mouse ESCs, at least in part, by down-regulating the expression of the anti-stemness microRNA (miRNA) miR-24-2. To gain greater insight into the molecular basis underlying PRMT7-mediated maintenance of mouse ESC stemness, we searched for new PRMT7-down-regulated anti-stemness miRNAs. Here, we show that miR-221 gene-encoded miR-221-3p and miR-221-5p are anti-stemness miRNAs whose expression levels in mouse ESCs are directly repressed by PRMT7. Notably, both miR-221-3p and miR-221-5p targeted the 3' untranslated regions of mRNA transcripts of the major pluripotency factors Oct4, Nanog, and Sox2 to antagonize mouse ESC stemness. Moreover, miR-221-5p silenced also the expression of its own transcriptional repressor PRMT7. Transfection of miR-221-3p and miR-221-5p mimics induced spontaneous differentiation of mouse ESCs. CRISPR-mediated deletion of the miR-221 gene, as well as specific antisense inhibitors of miR-221-3p and miR- 221-5p, inhibited the spontaneous differentiation of PRMT7- depleted mouse ESCs. Taken together, these findings reveal that the PRMT7-mediated repression of miR-221-3p and miR- 221-5p expression plays a critical role in maintaining mouse ESC stemness. Our results also establish miR-221-3p and miR- 221-5p as anti-stemness miRNAs that target Oct4, Nanog, and Sox2 mRNAs in mouse ESCs.
AB - The stemness maintenance of embryonic stem cells (ESCs) requires pluripotency transcription factors, including Oct4, Nanog, and Sox2. We have previously reported that protein arginine methyltransferase 7 (PRMT7), an epigenetic modifier, is an essential pluripotency factor that maintains the stemness of mouse ESCs, at least in part, by down-regulating the expression of the anti-stemness microRNA (miRNA) miR-24-2. To gain greater insight into the molecular basis underlying PRMT7-mediated maintenance of mouse ESC stemness, we searched for new PRMT7-down-regulated anti-stemness miRNAs. Here, we show that miR-221 gene-encoded miR-221-3p and miR-221-5p are anti-stemness miRNAs whose expression levels in mouse ESCs are directly repressed by PRMT7. Notably, both miR-221-3p and miR-221-5p targeted the 3' untranslated regions of mRNA transcripts of the major pluripotency factors Oct4, Nanog, and Sox2 to antagonize mouse ESC stemness. Moreover, miR-221-5p silenced also the expression of its own transcriptional repressor PRMT7. Transfection of miR-221-3p and miR-221-5p mimics induced spontaneous differentiation of mouse ESCs. CRISPR-mediated deletion of the miR-221 gene, as well as specific antisense inhibitors of miR-221-3p and miR- 221-5p, inhibited the spontaneous differentiation of PRMT7- depleted mouse ESCs. Taken together, these findings reveal that the PRMT7-mediated repression of miR-221-3p and miR- 221-5p expression plays a critical role in maintaining mouse ESC stemness. Our results also establish miR-221-3p and miR- 221-5p as anti-stemness miRNAs that target Oct4, Nanog, and Sox2 mRNAs in mouse ESCs.
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U2 - 10.1074/jbc.RA117.000425
DO - 10.1074/jbc.RA117.000425
M3 - Article
C2 - 29378844
AN - SCOPUS:85044115524
SN - 0021-9258
VL - 293
SP - 3925
EP - 3936
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 11
ER -