Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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The Study on Function and Localization of Nup97 in Fission Yeast

분열효모에서 Nup97의 기능과 세포 내 위치에 대한 연구

  • Hwang, Duk-Kyung (Basic Science Research Institute and Department of Biology, College of Natural Sciences, Sungshin Women's University) ;
  • Yoon, Jin-Ho (Basic Science Research Institute and Department of Biology, College of Natural Sciences, Sungshin Women's University)
  • 황덕경 (성신여자대학교 자연과학대학 생물학과 및 기초과학연구소) ;
  • 윤진호 (성신여자대학교 자연과학대학 생물학과 및 기초과학연구소)
  • Published : 2008.06.30
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Abstract

We studied on the function and localization of fission yeast Schizosaccharomyces pombe Nup97p, which is homologous to nucleoporin Nic96p in budding yeast Saccharomyces cerevisiae. There was no effect on growth and $poly(A)^{+}$ RNA distribution of cells when nup97 gene was overexpressed. However, the haploid ${\Delta}nup97::kan^{r}$ null mutants confirmed extensive $poly(A)^{+}$ RNA accumulation in the nucleus, abnormal DNA distribution, and cessation of growth when nup97 expression was repressed. We determined the subcellular localization of Nup97 tagged at the N terminus or the C terminus with GFP. Both fusions complemented growth defect of ${\Delta}nup97::kan^{r}$ null mutants. An integrated version of the nup97-GFP fusion was constructed at the nup97 locus. Nup97-GFP fusions expressed from its own promoter was localized at the nuclear periphery with a punctate appearance. These results suggest that Nup97p in fission yeast is also nucleoporin, which is involved in mRNA export.

발아효모 Saccharomyces cerevisiae의 핵공단백질인 Nic96 단백질과 유사성을 보이는 분열효모 Schizosaccharomyces pombe의 Nup97의 기능과 세포 내 위치를 조사하였다. nup97을 과 발현시켰을 때는 생장과 $poly(A)^{+}$ RNA의 분포에 별다른 이상을 보이지 않았다. 하지만, $kan^{r}$ 유전자를 이용하여 제작한 ${\Delta}nup97$ 결실돌연변이는 nup97의 발현이 억제되면, $poly(A)^{+}$ RNA가 핵 안에 축적되었고 비정상적인 DNA 분포를 보였으며 결국 생장하지 못했다. 한편, Nup97p의 N말단 또는 C말단에 GFP를 붙인 Nup97-GFP 융합단배질의 세포 내 위치를 확인하고자 하였다. 이러한 융합단백질들이 ${\Delta}nup97$ 결실돌연변이의 생장결함을 정상적으로 상보하는 것을 확인하고, nup97-GFP 유전자를 염색체의 nup97 유전자 위치에 삽입한 균주를 제작하였다. 자신의 프로모터에서 발현된 Nup97-GFP 융합단백질은 정상적인 기능을 보였으며, 핵막 주위에 점점 이 위치하였다. 이와 같은 결과들은 Nup97 단백질 역시 핵공단배질로 mRNA의 핵에서 세포질로의 이동에 중요하다는 것을 시사한다.

Keywords

References

  1. Alfa, C., P. Fantes, J. Hyams, M. Mcleod, and E. Warbrick. 1993. Experiments with Fission Yeast. Cold Spring Harbor, Cold Spring Harbor Laboratory, New York, NY, USA
  2. Cho, H.J., D.K. Hwang, S.I. Jung, and J.H. Yoon. 2007. Schizosaccharomyces pombe nup97, which genetically interacts with mex67, is essential for growth and involved in mRNA export. J. Microbiol. 45, 344-349
  3. Cronshaw, J.M., A.N. Krutchinsky, W. Zhang, B.T. Chait, and M.J. Matunis. 2002. Proteomic analysis of the mammalian nuclear pore complex. J. Cell Biol. 158, 915-927 https://doi.org/10.1083/jcb.200206106
  4. Cullen, B.R. 2003. Nuclear RNA export. J. Cell Sci. 116, 587-597 https://doi.org/10.1242/jcs.00268
  5. Erkmann, J.A. and U. Kutay. 2004. Nuclear export of mRNA: from the site of transcription of the cytoplasm. Exp. Cell Res. 296, 12-20 https://doi.org/10.1016/j.yexcr.2004.03.015
  6. Fahrenkrog, B. and U. Aebi. 2003. The nuclear pore complex: nucleocytoplasmic transport and beyond. Nat. Rev. Mol. Cell Biol. 4, 757-766 https://doi.org/10.1038/nrm1230
  7. Grlich, D. and U. Kutay. 1999. Transport between the cell nucleus and the cytoplasm. Annu. Rev. Cell Dev. Biol. 15, 607-660 https://doi.org/10.1146/annurev.cellbio.15.1.607
  8. Hetzer, M.W., T.C. Walther, and I.W. Mattaj. 2005. Pushing the envelope: structure, function, and dynamics of the nuclear periphery. Annu. Rev. Cell Dev. Biol. 21, 347-380 https://doi.org/10.1146/annurev.cellbio.21.090704.151152
  9. Izaurralde, E. 2002. A novel family of nuclear transport receptors mediates the export of messenger RNA to the cytoplasm. Eur. J. Cell Biol. 81, 577-584 https://doi.org/10.1078/0171-9335-00273
  10. Lei, E.P. and P.A. Silver. 2002. Protein and RNA export from the nucleus. Dev. Cell 2, 261-272 https://doi.org/10.1016/S1534-5807(02)00134-X
  11. Moore, M.S. and G.A. Blobel. 1994. G protein involved in nucleocytoplasmic transport: the role of Ran. Trends Biochem. Sci. 19, 211-216 https://doi.org/10.1016/0968-0004(94)90024-8
  12. Moreno, S., A. Klar, and P. Nurse. 1991. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol. 194, 795-823 https://doi.org/10.1016/0076-6879(91)94059-L
  13. Reed, R. and E. Hurt. 2002. A conserved mRNA export machinery coupled to pre-mRNA splicing. Cell 108, 523-531 https://doi.org/10.1016/S0092-8674(02)00627-X
  14. Rout, M.P., J.D. Aitchison, A. Suprapto, K. Hjertaas, Y. Zhao, and B.T. Chait. 2000. The yeast nuclear pore complex: composition, architecture, and transport mechanism. J. Cell. Biol. 148, 635-651 https://doi.org/10.1083/jcb.148.4.635
  15. Suntharalingam, M. and S.R. Wente. 2003. Peering through the pore: nuclear pore complex structure, assembly, and function. Dev. Cell 4, 775-789 https://doi.org/10.1016/S1534-5807(03)00162-X
  16. Trans, E.J. and S.R. Wente. 2006. Dynamic nuclear pore complexes: life on the edge. Cell 125, 1041-1053 https://doi.org/10.1016/j.cell.2006.05.027
  17. Vinciguerra, P. and F. Stutz. 2004. mRNA export: an assembly line from genes to nuclear pores. Curr. Opin. Cell Biol. 16, 285-292 https://doi.org/10.1016/j.ceb.2004.03.013
  18. Weis, K. 2002. Nucleocytoplasmic transport: cargo trafficking across the border. Curr. Opin. Cell Biol. 14, 328-335 https://doi.org/10.1016/S0955-0674(02)00337-X
  19. Yoon, J.H. 2003. Synthetic lethal mutations with spmex67 of Schizosaccharomyces pombe in the mediation of mRNA export. J. Microbiol. 41, 115-120
  20. Yoon, J.H., D. Love, A. Guhathakurta, J.A. Hanover, and R. Dhar. 2000. Mex67p of Schizosaccharomyces pombe interacts with Rae1p in mediating mRNA export. Mol. Cell. Biol. 20, 8767-8782 https://doi.org/10.1128/MCB.20.23.8767-8782.2000
  21. Yu, J.H., Z. Hamari, K.H. Han, J.A. Seo, Y. Reyes-Domnguez, and C. Scazzocchio. 2004. Double-joint PCR: a PCR-based molecular fool for gene manipulations in filamentous fungi. Fungal Genet. Biol. 41, 973-981 https://doi.org/10.1016/j.fgb.2004.08.001