DOI QR코드

DOI QR Code

Zebrafish Dnd protein binds to 3'UTR of geminin mRNA and regulates its expression

  • Chen, Shu (Department of Medical Genetics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University) ;
  • Zeng, Mei (Department of Medical Genetics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University) ;
  • Sun, Huaqin (Department of Medical Genetics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University) ;
  • Deng, Wenqian (Department of Medical Genetics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University) ;
  • Lu, Yilu (Department of Medical Genetics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University) ;
  • Tao, Dachang (Department of Medical Genetics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University) ;
  • Liu, Yunqiang (Department of Medical Genetics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University) ;
  • Zhang, Sizhong (Department of Medical Genetics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University) ;
  • Ma, Yongxin (Department of Medical Genetics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University)
  • Received : 2010.04.29
  • Accepted : 2010.05.01
  • Published : 2010.06.30

Abstract

Dnd (dead end) gene encodes an RNA binding protein and is specifically expressed in primordial germ cells (PGCs) as a vertebrate-specific component of the germ plasma throughout embryogenesis. By utilizing a technique of specific nucleic acids associated with proteins (SNAAP), 13 potential target mRNAs of zebrafish Dnd (ZDnd) protein were identified from 8-cell embryo, and 8 target mRNAs have been confirmed using an RT-PCR analysis. Of the target mRNAs, the present study is focused on the regulation of geminin, which is an inhibitor of DNA replication. Using electrophoretic mobility shift assay (EMSA), we demonstrated that ZDND protein bound the 67-nucleotide region from 864 to 931 in the 3'UTR of geminin mRNA, a sequence containing 60.29% of uridine. Results from a dual-luciferase assay in HEK293 cells showed that ZDND increases the translation of geminin. Taken together, the identification of target mRNA for ZDnd will be helpful to further explore the biological function of Dnd in zebrafish germ-line development as well as in cancer cells.

Keywords

References

  1. Glisovic, T., Bachorik, J. L., Yong, J. and Dreyfuss, G. (2008) RNA-binding proteins and post-transcriptional gene regulation. FEBS Lett. 582, 1977-1986. https://doi.org/10.1016/j.febslet.2008.03.004
  2. Venables, J. P. and Eperon, I. (1999) The roles of RNAbinding proteins in spermatogenesis and male infertility. Curr. Opin. Genet. Dev. 9, 346-354. https://doi.org/10.1016/S0959-437X(99)80052-5
  3. Chen, X., Li, W. Z., Shao, Y. and QC, Z. (2007) Progress in RNA-binding proteins in animals and plants. Biotechnology Bulletin 3, 7.
  4. Horvay, K., Claussen, M., Katzer, M., Landgrebe, J. and Pieler, T. (2006) Xenopus Dead end mRNA is a localized maternal determinant that serves a conserved function in germ cell development. Dev. Biol. 291, 1-11. https://doi.org/10.1016/j.ydbio.2005.06.013
  5. Youngren, K. K., Coveney, D., Peng, X., Bhattacharya, C., Schmidt, L. S., Nickerson, M. L., Lamb, B. T., Deng, J. M., Behringer, R. R., Capel, B., Rubin, E. M., Nadeau, J. H. and Matin, A. (2005) The Ter mutation in the dead end gene causes germ cell loss and testicular germ cell tumours. Nature 435, 360-364. https://doi.org/10.1038/nature03595
  6. Weidinger, G., Stebler, J., Slanchev, K., Dumstrei, K., Wise, C., Lovell-Badge, R., Thisse, C., Thisse, B. and Raz, E. (2003) dead end, a novel vertebrate germ plasm component, is required for zebrafish primordial germ cell migration and survival. Curr. Biol. 13, 1429-1434. https://doi.org/10.1016/S0960-9822(03)00537-2
  7. Liu, L., Hong, N., Xu, H., Li, M., Yan, Y., Purwanti, Y., Yi, M., Li, Z., Wang, L. and Hong, Y. (2009) Medaka dead end encodes a cytoplasmic protein and identifies embryonic and adult germ cells. Gene. Expr. Patterns. 9, 541-548. https://doi.org/10.1016/j.gep.2009.06.008
  8. Aramaki, S., Sato, F., Kato, T., Soh, T., Kato, Y. and Hattori, M. A. (2007) Molecular cloning and expression of dead end homologue in chicken primordial germ cells. Cell Tissue. Res. 330, 45-52. https://doi.org/10.1007/s00441-007-0435-1
  9. Lam, M. Y., Heaney, J. D., Youngren, K. K., Kawasoe, J. H. and Nadeau, J. H. (2007) Trans-generational epistasis between Dnd1Ter and other modifier genes controls susceptibility to testicular germ cell tumors. Hum. Mol. Genet. 16, 2233-2240. https://doi.org/10.1093/hmg/ddm175
  10. Linger, R., Dudakia, D., Huddart, R., Tucker, K., Friedlander, M., Phillips, K. A., Hogg, D., Jewett, M. A., Lohynska, R., Daugaard, G., Richard, S., Chompret, A., Stoppa-Lyonnet, D., Bonaiti-Pellie, C., Heidenreich, A., Albers, P., Olah, E., Geczi, L., Bodrogi, I., Daly, P. A., Guilford, P., Fossa, S. D., Heimdal, K., Tjulandin, S. A., Liubchenko, L., Stoll, H., Weber, W., Einhorn, L., McMaster, M., Korde, L., Greene, M. H., Nathanson, K. L., Cortessis, V., Easton, D. F., Bishop, D. T., Stratton, M. R. and Rapley, E. A. (2008) Analysis of the DND1 gene in men with sporadic and familial testicular germ cell tumors. Genes Chromosomes Cancer 47, 247-252. https://doi.org/10.1002/gcc.20526
  11. Kedde, M., Strasser, M. J., Boldajipour, B., Oude Vrielink, J. A., Slanchev, K., le Sage, C., Nagel, R., Voorhoeve, P. M., van Duijse, J., Orom, U. A., Lund, A. H., Perrakis, A., Raz, E. and Agami, R. (2007) RNA-binding protein Dnd1 inhibits microRNA access to target mRNA. Cell 131, 1273-1286. https://doi.org/10.1016/j.cell.2007.11.034
  12. Trifillis, P., Day, N., and Kiledjian, M. (1999) Finding the right RNA: identification of cellular mRNA substrates for RNA-binding proteins. RNA 5, 1071-1082. https://doi.org/10.1017/S1355838299981803
  13. McGarry, T. J. and Kirschner, M. W. (1998) Geminin, an inhibitor of DNA replication, is degraded during mitosis. Cell 93, 1043-1053. https://doi.org/10.1016/S0092-8674(00)81209-X
  14. Pitulescu, M., Kessel, M. and Luo, L. (2005) The regulation of embryonic patterning and DNA replication by geminin. Cell Mol. Life Sci. 62, 1425-1433. https://doi.org/10.1007/s00018-005-4553-1
  15. Luo, L., Uerlings, Y., Happel, N., Asli, N. S., Knoetgen, H. and Kessel, M. (2007) Regulation of geminin functions by cell cycle-dependent nuclear-cytoplasmic shuttling. Mol. Cell Biol. 27, 4737-4744. https://doi.org/10.1128/MCB.00123-07
  16. Tachibana, K. E., Gonzalez, M. A., Guarguaglini, G., Nigg, E. A. and Laskey, R. A. (2005) Depletion of licensing inhibitor geminin causes centrosome overduplication and mitotic defects. EMBO Rep. 6, 1052-1057. https://doi.org/10.1038/sj.embor.7400527
  17. Xouri, G., Lygerou, Z., Nishitani, H., Pachnis, V., Nurse, P. and Taraviras, S. (2004) Cdt1 and geminin are downregulated upon cell cycle exit and are over-expressed in cancer-derived cell lines. Eur. J. Biochem. 271, 3368-3378. https://doi.org/10.1111/j.1432-1033.2004.04271.x
  18. Westerfield, M. (1993) The Zebrafish Book. University of Oregon Press, Oregon, USA.
  19. Kimmel, C. B., Ballard, W. W., Kimmel, S. R., Ullmann, B. and Schilling, T. F. (1995) Stages of embryonic development of the zebrafish. Dev. Dyn. 203, 253-310. https://doi.org/10.1002/aja.1002030302
  20. Jiao, X., Trifillis, P. and Kiledjian, M. (2002) Identification of target messenger RNA substrates for the murine deleted in azoospermia-like RNA-binding protein. Biol. Reprod. 66, 475-485. https://doi.org/10.1095/biolreprod66.2.475
  21. Paronetto, M. P., Achsel, T., Massiello, A., Chalfant, C. E. and Sette, C. (2007) The RNA-binding protein Sam68 modulates the alternative splicing of Bcl-x. J. Cell Biol. 176, 929-939. https://doi.org/10.1083/jcb.200701005
  22. King, P. H. (2000) RNA-binding analyses of HuC and HuD with the VEGF and c-myc 3'-untranslated regions using a novel ELISA-based assay. Nucleic. Acids. Res. 28, E20. https://doi.org/10.1093/nar/28.7.e20

Cited by

  1. Germ cell-specific expression of dead end (dnd) in rare minnow (Gobiocypris rarus) vol.41, pp.2, 2015, https://doi.org/10.1007/s10695-015-0029-x
  2. Geminin a multi task protein involved in cancer pathophysiology and developmental process: A review vol.131, 2016, https://doi.org/10.1016/j.biochi.2016.09.022
  3. In Vitro Generation of Zebrafish PGC-Like Cells1 vol.91, pp.5, 2014, https://doi.org/10.1095/biolreprod.114.121491
  4. The germ cell marker dead end reveals alternatively spliced transcripts with dissimilar expression vol.9, pp.1, 2019, https://doi.org/10.1038/s41598-019-39101-9