Journal of Animal Reproduction and Biotechnology (한국동물생명공학회지)

The Korean Society of Animal Reproduction and Biotechnology (한국동물생명공학회)
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M6A reader hnRNPA2/B1 is essential for porcine embryo development via gene expression regulation

  • Kwon, Jeongwoo (Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Jo, Yu-Jin (Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Yoon, Seung-Bin (Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • You, Hyeong-ju (Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Youn, Changsic (Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Yejin (Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Lee, Jiin (Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Nam-Hyung (Guangdong Provincial Key Laboratory of Large Animal models for Biomedicine, Wuyi university) ;
  • Kim, Ji-Su (Primate Resources Center (PRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB))
  • Received : 2022.06.08
  • Accepted : 2022.06.22
  • Published : 2022.06.30
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Abstract

Heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNPA2/B1) is an N6-methyladenosine (m6A) RNA modification regulator and a key determinant of prem-RNA processing, mRNA metabolism and transportation in cells. Currently, m6A reader proteins such as hnRNPA2/B1 and YTHDF2 has functional roles in mice embryo. However, the role of hnRNPA2/B1 in porcine embryogenic development are unclear. Here, we investigated the developmental competence and mRNA expression levels in porcine parthenogenetic embryos after hnRNPA2/B1 knock-down. HhnRNPA2/B1 was localized in the nucleus during subsequent embryonic development since zygote stage. After hnRNPA2/B1 knock-down using double stranded RNA injection, blastocyst formation rate decreased than that in the control group. Moreover, hnRNPA2/B1 knock-down embryos show developmental delay after compaction. In blastocyste stage, total cell number was decreased. Interestingly, gene expression patterns revealed that transcription of Pou5f1, Sox2, TRFP2C, Cdx2 and PARD6B decreased without changing the junction protein, ZO1, OCLN, and CDH1. Thus, hnRNPA2/B1 is necessary for porcine early embryo development by regulating gene expression through epigenetic RNA modification.

Keywords

Acknowledgement

This study was supported by grants from the KRIBB Research Initiative Program (KGM5162221).

References

  1. Alarcon CR, Goodarzi H, Lee H, Liu X, Tavazoie S, Tavazoie SF. 2015. HNRNPA2B1 is a mediator of m(6)A-dependent nuclear RNA processing events. Cell 162:1299-1308. https://doi.org/10.1016/j.cell.2015.08.011
  2. Aoki F, Worrad DM, Schultz RM. 1997. Regulation of transcriptional activity during the first and second cell cycles in the preimplantation mouse embryo. Dev. Biol. 181:296-307. https://doi.org/10.1006/dbio.1996.8466
  3. Apostolou E and Hochedlinger K. 2013. Chromatin dynamics during cellular reprogramming. Nature 502:462-471. https://doi.org/10.1038/nature12749
  4. Batista PJ, Molinie B, Wang J, Qu K, Zhang J, Li L, Bouley DM, Lujan E, Haddad B, Daneshvar K, Carter AC, Flynn RA, Zhou C, Lim KS, Dedon P, Wernig M, Mullen AC, Xing Y, Giallourakis CC, Chang HY. 2014. m(6)A RNA modification controls cell fate transition in mammalian embryonic stem cells. Cell Stem Cell 15:707-719. https://doi.org/10.1016/j.stem.2014.09.019
  5. Beyer AL, Christensen ME, Walker BW, LeStourgeon WM. 1977. Identification and characterization of the packaging proteins of core 40S hnRNP particles. Cell 11:127-138. https://doi.org/10.1016/0092-8674(77)90323-3
  6. Bokar JA, Shambaugh ME, Polayes D, Matera AG, Rottman FM. 1997. Purification and cDNA cloning of the AdoMet-binding subunit of the human mRNA (N6-adenosine)-methyltransferase. RNA 3:1233-1247.
  7. Burton A and Torres-Padilla ME. 2014. Chromatin dynamics in the regulation of cell fate allocation during early embryogenesis. Nat. Rev. Mol. Cell Biol. 15:723-734. https://doi.org/10.1038/nrm3885
  8. Chang H, Lim J, Ha M, Kim VN. 2014. TAIL-seq: genome-wide determination of poly(A) tail length and 3' end modifications. Mol. Cell 53:1044-1052. https://doi.org/10.1016/j.molcel.2014.02.007
  9. Choi HS, Lee HM, Jang YJ, Kim CH, Ryu CJ. 2013. Heterogeneous nuclear ribonucleoprotein A2/B1 regulates the selfrenewal and pluripotency of human embryonic stem cells via the control of the G1/S transition. Stem Cells 31:2647-2658. https://doi.org/10.1002/stem.1366
  10. Desrosiers RC, Friderici KH, Rottman FM. 1975. Characterization of Novikoff hepatoma mRNA methylation and heterogeneity in the methylated 5' terminus. Biochemistry 14:4367-4374. https://doi.org/10.1021/bi00691a004
  11. Dreyfuss G, Matunis MJ, Pinol-Roma S, Burd CG. 1993. hnRNP proteins and the biogenesis of mRNA. Annu. Rev. Biochem. 62:289-321. https://doi.org/10.1146/annurev.bi.62.070193.001445
  12. Fawcett KA and Barroso I. 2010. The genetics of obesity: FTO leads the way. Trends Genet. 26:266-274. https://doi.org/10.1016/j.tig.2010.02.006
  13. Hanna JH, Saha K, Jaenisch R. 2010. Pluripotency and cellular reprogramming: facts, hypotheses, unresolved issues. Cell 143:508-525. https://doi.org/10.1016/j.cell.2010.10.008
  14. Hyttel P, Laurincik J, Rosenkranz Ch, Rath D, Niemann H, Ochs RL, Schellander K. 2000. Nucleolar proteins and ultrastructure in preimplantation porcine embryos developed in vivo. Biol. Reprod. 63:1848-1856. https://doi.org/10.1095/biolreprod63.6.1848
  15. Ivanova I, Much C, Di Giacomo M, Azzi C, Morgan M, Moreira PN, Monahan J, Carrieri C, Enright AJ, O'Carroll D. 2017. The RNA m6A reader YTHDF2 is essential for the post-transcriptional regulation of the maternal transcriptome and oocyte competence. Mol. Cell 67:1059-1067.e4. https://doi.org/10.1016/j.molcel.2017.08.003
  16. Kashyap V, Rezende NC, Scotland KB, Shaffer SM, Persson JL, Gudas LJ, Mongan NP. 2009. Regulation of stem cell pluripotency and differentiation involves a mutual regulatory circuit of the NANOG, OCT4, and SOX2 pluripotency transcription factors with polycomb repressive complexes and stem cell microRNAs. Stem Cells Dev. 18:1093-1108. https://doi.org/10.1089/scd.2009.0113
  17. Kwon J, Jo YJ, Namgoong S, Kim NH. 2019. Functional roles of hnRNPA2/B1 regulated by METTL3 in mammalian embryonic development. Sci. Rep. 9:8640. https://doi.org/10.1038/s41598-019-44714-1
  18. Latham KE, Garrels JI, Chang C, Solter D. 1991. Quantitative analysis of protein synthesis in mouse embryos. I. Extensive reprogramming at the one- and two-cell stages. Development 112:921-932. https://doi.org/10.1242/dev.112.4.921
  19. Liu J, Yue Y, Han D, Wang X, Fu Y, Zhang L, Jia G, Yu M, Lu Z, Deng X, Dai Q, Chen W, He C. 2014. A METTL3-METTL14 complex mediates mammalian nuclear RNA N6-adenosine methylation. Nat. Chem. Biol. 10:93-95. https://doi.org/10.1038/nchembio.1432
  20. Perry RP, Kelley DE, Friderici K, Rottman F. 1975. The methylated constituents of L cell messenger RNA: evidence for an unusual cluster at the 5' terminus. Cell 4:387-394. https://doi.org/10.1016/0092-8674(75)90159-2
  21. Tadros W and Lipshitz HD. 2009. The maternal-to-zygotic transition: a play in two acts. Development 136:3033-3042. https://doi.org/10.1242/dev.033183
  22. Tang C, Klukovich R, Peng H, Wang Z, Yu T, Zhang Y, Zheng H, Klungland A, Yan W. 2018. ALKBH5-dependent m6A demethylation controls splicing and stability of long 3'-UTR mRNAs in male germ cells. Proc. Natl. Acad. Sci. U. S. A. 115: E325-E333.
  23. Telford NA, Watson AJ, Schultz GA. 1990. Transition from maternal to embryonic control in early mammalian development: a comparison of several species. Mol. Reprod. Dev. 26:90-100. https://doi.org/10.1002/mrd.1080260113
  24. Ulitsky I, Shkumatava A, Jan CH, Subtelny AO, Koppstein D, Bell GW, Sive H, Bartel DP. 2012. Extensive alternative polyadenylation during zebrafish development. Genome Res. 22:2054-2066. https://doi.org/10.1101/gr.139733.112
  25. Wang G, Xiao Q, Luo Z, Ye S, Xu Q. 2012. Functional impact of heterogeneous nuclear ribonucleoprotein A2/B1 in smooth muscle differentiation from stem cells and embryonic arteriogenesis. J. Biol. Chem. 287:2896-2906. https://doi.org/10.1074/jbc.M111.297028
  26. Wang X, Lu Z, Gomez A, Hon GC, Yue Y, Han D, Fu Y, Parisien M, Dai Q, Jia G, Ren B, Pan T, He C. 2014. N6-methyladenosine-dependent regulation of messenger RNA stability. Nature 505:117-120. https://doi.org/10.1038/nature12730
  27. Wang X, Zhao BS, Roundtree IA, Lu Z, Han D, Ma H, Weng X, Chen K, Shi H, He C. 2015. N(6)-methyladenosine modulates messenger RNA translation efficiency. Cell 161:1388-1399. https://doi.org/10.1016/j.cell.2015.05.014
  28. Wei CM, Gershowitz A, Moss B. 1975. Methylated nucleotides block 5' terminus of HeLa cell messenger RNA. Cell 4:379-386. https://doi.org/10.1016/0092-8674(75)90158-0
  29. Zhao BS, Wang X, Beadell AV, Lu Z, Shi H, Kuuspalu A, Ho RK, He C. 2017. M6A-dependent maternal mRNA clearance facilitates zebrafish maternal-to-zygotic transition. Nature 542: 475-478. https://doi.org/10.1038/nature21355
  30. Zhao BS and He C. 2017. "Gamete on" for m6A: YTHDF2 exerts essential functions in female fertility. Mol. Cell 67:903-905. https://doi.org/10.1016/j.molcel.2017.09.004
  31. Zheng G, Dahl JA, Niu Y, Fedorcsak P, Huang CM, Li CJ, Vagbo CB, Shi Y, Wang WL, Song SH, Lu Z, Bosmans RP, Dai Q, Hao YJ, Yang X, Zhao WM, Tong WM, Wang XJ, Bogdan F, Furu K, Fu Y, Jia G, Zhao X, Liu J, Krokan HE, Klungland A, Yang YG, He C. 2013. ALKBH5 is a mammalian RNA demethylase that impacts RNA metabolism and mouse fertility. Mol. Cell 49: 18-29. https://doi.org/10.1016/j.molcel.2012.10.015