Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
권호 표지

Schizosaccharomyces pombe nup97, which Genetically Interacts with mex67, is Essential for Growth and Involved in mRNA Export

  • Cho, Hyun-Jin (Department of Biology, Institute of Basic Sciences, Sungshin Women's University) ;
  • Hwang, Duk-Kyung (Department of Biology, Institute of Basic Sciences, Sungshin Women's University) ;
  • Jung, Sun-Im (Department of Biology, Institute of Basic Sciences, Sungshin Women's University) ;
  • Yoon, Jin-Ho (Department of Biology, Institute of Basic Sciences, Sungshin Women's University)
  • Published : 2007.08.30
Download PDF
⟨ Previous Next ⟩

Abstract

We have isolated previously three synthetic lethal mutants in Schizosaccharomyces pombe, which genetically interact with mex67, in order to identify the genes involved in mRNA export. A novel nup97 gene was isolated by complementation of the growth defect in one of the synthetic lethal mutants, SLMex3. The nup97 gene contains one intron and encodes an 851 amino-acid protein that is similar to nucleoporins, Nppl06p in S. pombe and Nic96p in Saccharomyces cerevisiae. The nup97 gene is essential for vegetative growth, and nup97 null mutant harboring pREP41X-Nup97 showed $poly(A)^+$ RNA export defect when expression of nup97 is repressed in the presence of thiamine. These results suggest that nup97 is involved in mRNA export from the nucleus to cytoplasm.

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, NY, USA
  2. Barbet, N., W.J. Muriel, and A.M. Carr. 1992. Versatile shuttle vectors and genomic libraries for use with Schizosaccharomyces pombe. Gene 114, 59-66 https://doi.org/10.1016/0378-1119(92)90707-V
  3. Basi, G., E. Schmid, and K. Maundrell. 1993. TATA box mutations in the Schizosaccharomyces pombe nmt1 promoter affect transcription efficiency but not the transcription start point or thiamine repressibility. Gene 123, 131-136 https://doi.org/10.1016/0378-1119(93)90552-E
  4. Braun, I.C., A. Herold, M. Rode, and E. Izaurralde. 2002. Nuclear export of mRNA by TAP/NXF1 requires two nucleoporinbinding sites but not p15. Mol. Cell. Biol. 22, 5405-5418 https://doi.org/10.1128/MCB.22.15.5405-5418.2002
  5. Brown, J.A., A. Bharathi, A. Ghosh, W. Whalen, E. Fitzgerald, and R. Dhar. 1995. A mutation in the Schizosaccharomyces pombe rae1 gene causes defects in $poly(A)^+$ RNA export and in the cytoskeleton. J. Biol. Chem. 270, 7411-7419 https://doi.org/10.1074/jbc.270.13.7411
  6. Cole, C.N. and J.J. Scarcelli. 2006. Unravelling mRNA export. Nat. Cell. Biol. 8, 645-647 https://doi.org/10.1038/ncb0706-645
  7. 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
  8. Devos, D., S. Dokudovskaya, F. Alber, R. Williams, B.T. Chait, A. Sali, and M.P. Rout. 2004. Components of coated vesicles and nuclear pore complexes share a common molecular architecture. PLoS Biol. 2, e380 https://doi.org/10.1371/journal.pbio.0020380
  9. Devos, D., S. Dokudovskaya, R. Williams, F. Alber, N. Eswar, B.T. Chait, M.P. Rout, and A. Sali. 2006. Simple fold composition and modular architecture of the nuclear pore complex. Proc. Natl. Acad. Sci. USA 103, 2172-2177
  10. Erkmann, J.A. and U. Kutay. 2004. Nuclear export of mRNA: from the site of transcription to the cytoplasm. Exp. Cell Res. 296, 12-20 https://doi.org/10.1016/j.yexcr.2004.03.015
  11. Fahrenkrog, B., E.C. Hurt, U. Aebi, and N. Pante. 1998. Molecular architecture of the yeast nuclear pore complex: localization of Nsp1p subcomplexes. J. Cell Biol. 143, 577-588 https://doi.org/10.1083/jcb.143.3.577
  12. Forsburg, S.L. 1993. Comparison of Schizosaccharomyces pombe expression systems. Nucleic Acids Res. 21, 2955-2956 https://doi.org/10.1093/nar/21.12.2955
  13. Grandi, P., V. Doye, and E.C. Hurt. 1993. Purification of NSP1 reveals complex formation with 'GLFG' nucleoporins and a novel nuclear pore protein NIC96. EMBO J. 12, 3061-3071
  14. Grandi, P., N. Schlaich, H. Tekotte, and E.C. Hurt. 1995. Functional interaction of Nic96p with a core nucleoporin complex consisting of Nsp1p, Nup49p and a novel protein Nup57p. EMBO J. 14, 76-87
  15. Gwizdek, C., N. Iglesias, M.S. Rodriguez, B. Ossareh-Nazari, M. Hobeika, G. Divita, F. Stutz, and C. Dargemont. 2006. Ubiquitin- associated domain of Mex67 synchronizes recruitment of the mRNA export machinery with transcription. Proc. Natl. Acad. Sci. USA 103, 16376-16381
  16. Huang, Y. and J.A. Steitz. 2005. SRprises along a messenger's journey. Mol. Cell. 17, 613-615 https://doi.org/10.1016/j.molcel.2005.02.020
  17. Kozak, L., G. Gopal, J.H. Yoon, Z.E. Sauna, S.V. Ambudkar, A.G. Thakurta, and R. Dhar. 2002. Elf1p, a member of the ABC class of ATPases, functions as a mRNA export factor in Schizosacchromyces pombe. J. Biol. Chem. 277, 33580-33589 https://doi.org/10.1074/jbc.M205415200
  18. Matsumoto, T. and D. Beach. 1991. Premature initiation of mitosis in yeast lacking RCC1 or an interacting GTPase. Cell 66, 347-360 https://doi.org/10.1016/0092-8674(91)90624-8
  19. Maundrell, K. 1993. Thiamine-repressible expression vectors pREP and pRIP for fission yeast. Gene 123, 127-130 https://doi.org/10.1016/0378-1119(93)90551-D
  20. Moore, M.J. 2005. From birth to death: the complex lives of eukaryotic mRNAs. Science 309, 1514-1518 https://doi.org/10.1126/science.1111443
  21. 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
  22. 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
  23. Rodriguez, M.S., C. Dargemont, and F. Stutz. 2004. Nuclear export of RNA. Biol. Cell 96, 639-655 https://doi.org/10.1016/j.biolcel.2004.04.014
  24. Rout, M.P. and J.D. Aitchison. 2001. The nuclear pore complex as a transport machine. J. Biol. Chem. 276, 16593-16596 https://doi.org/10.1074/jbc.R100015200
  25. 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
  26. Segref, A., K. Sharma, V. Doye, A. Hellwig, J. Huber, R. Luhrmann, and E. Hurt. 1997. Mex67p, a novel factor for nuclear mRNA export, binds to both $poly(A)^+$ RNA and nuclear pores. EMBO J. 16, 3256-3271 https://doi.org/10.1093/emboj/16.11.3256
  27. Stoffler, D., B. Fahrenkrog, and U. Aebi. 1999. The nuclear pore complex: from molecular architecture to functional dynamics. Curr. Opin. Cell Biol. 11, 391-401 https://doi.org/10.1016/S0955-0674(99)80055-6
  28. 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
  29. Thakurta, A.G., G. Gopal, J.H. Yoon, L. Kozak, and R. Dhar. 2005. Homolog of BRCA2-interacting Dss1p and Uap56p link Mlo3p and Rae1p for mRNA export in fission yeast. EMBO J. 24, 2512-2523 https://doi.org/10.1038/sj.emboj.7600713
  30. Tran, 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
  31. Vasu, S.K. and D.J. Forbes. 2001. Nuclear pores and nuclear assembly. Curr. Opin. Cell Biol. 13, 363-375 https://doi.org/10.1016/S0955-0674(00)00221-0
  32. Whalen, W.A., J.H. Yoon, R. Shen, and R. Dhar. 1999. Regulation of mRNA export by nutritional status in fission yeast. Genetics 152, 827-838
  33. Yoon, J.H. 2003. Synthetic lethal mutations with spmex67 of Schizosaccharomyces pombe in the mediation of mRNA export. J. Microbiol. 41, 115-120
  34. Yoon, J.H. 2004. Schizosaccharomyces pombe rsm1 genetically interacts with spmex67, which is involved in mRNA export. J. Microbiol. 42, 32-36
  35. Yoon, J.H., D.C. 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
  36. Yoon, J.H., W.A. Whalen, A. Bharathi, R. Shen, and R. Dhar. 1997. Npp106p, a Schizosaccharomyces pombe nucleoporin similar to Saccharomyces cerevisiae Nic96p, functionally interacts with Rae1p in mRNA export. Mol. Cell. Biol. 17, 7047-7060 https://doi.org/10.1128/MCB.17.12.7047
  37. Yu, J.H., Z. Hamari, K.H. Han, J.A. Seo, Y. Reyes-Dominguez, and C. Scazzocchio. 2004. Double-joint PCR: a PCR-based molecular tool for gene manipulations in filamentous fungi. Fungal Genet. Biol. 41, 973-981 https://doi.org/10.1016/j.fgb.2004.08.001
  38. Zabel, U., V. Doye, H. Tekotte, R. Wepf, P. Grandi, and E.C. Hurt. 1996. Nic96p is required for nuclear pore formation and functionally interacts with a novel nucleoporin, Nup188p. J. Cell Biol. 133, 1141-1152 https://doi.org/10.1083/jcb.133.6.1141