분열효모 Schizosaccharomyces pombe에서 spThp1 유전자 결실돌연변이의 제조와 특성 조사

Construction of Schizosaccharomyces pombe spThp1 Null Mutants and its Characterization

  • 윤진호 (성신여자대학교 생물학과 및 기초과학연구소)
  • Yoon Jin-Ho (Department of Biology and Institute of Basic Sciences, Sungshin Women's University)
  • 발행 : 2006.06.01

초록

분열효모인 Schizosaccharomyces pombe에서 mRNA의 핵에서 세포질로의 이동에 관여할 것으로 여겨지는 spThp1 유전자의 결실돌연변이주(deletion mutant)를 제조하여 그 특성을 조사하였다. 이배체(diploid) 균주의 한 spThp1 유전자를 결실시킨 후 4분체분석(tetrad analysis)을 수행한 결과, 이 유전자는 생장에 필수적이지 않았다. 또한 결실돌연변이주는 mRNA 수송도 큰 결함을 보이지 않았다. 하지만 spThp1 는 mRNA의 운반체를 암호화하고 있는 spMex67와 합성치사(synthetic lethality)를 보였다. 이 결과는 분열효모의 spThpl도 mRNA의 핵에서 세포질로의 이동에 역할을 하고 있음을 암시한다.

The sp%pl null mutant was constructed to study the function of fission yeast Schizosaccharomyces pombe spThp1, which is homologous to budding yeast Saccharomyces cerevisiae THP1. Tetrad analysis showed that the spThp1 is not essential for vegetative growth. The spThp1 null mutant also showed no massive poly(A)+ RNA export defect. However, spThp1 null is genetically associated with spMex67 null. These results suggest that spThp1 is involved in mRNA export out of the nucleus.

키워드

참고문헌

  1. Alfa, C., P. Fantes, J. Hyams, M. Mcleod, and E. Warbrick. 1993. Experiments with Fission Yeast. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory
  2. Aguilera. A. 2005. Cotranscriptional mRNP assembly: from the DNA to the nuclear pore. Curr. Opin. Cell Biol. 17, 242-250 https://doi.org/10.1016/j.ceb.2005.03.001
  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. Dreyfuss, G., V.N. Kim, and N. Kataoka. 2002. Messenger-RNAbinding proteins and the messages they carry. Nat. Rev. Mol. Cell Biol. 3, 195-205 https://doi.org/10.1038/nrm760
  6. 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
  7. Fischer, T., K. Strasser, A. Racz, S. Rodriguez-Navarro, M. Oppizzi, P. Ihrig, J. Lechner, and E. Hurt. 2002. The mRNA export machinery requires the novel Sac3p-Thp1p complex to dock at the nucleoplasmic entrance of the nuclear pores. EMBO J. 21, 5843-5852 https://doi.org/10.1093/emboj/cdf590
  8. Kang, Y. and B.R. Cullen. 1999. The human Tap protein is a nuclear mRNA export factor that contains novel RNA-binding and nucleocytoplasmic transport sequences. Genes Dev. 13(9); 1126-1139 https://doi.org/10.1101/gad.13.9.1126
  9. 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
  10. Lei, E.P., C.A. Stern, B. Fahrenkrog, H. Krebber, T.I. Moy, U. Aebi, and P.A. Silver. 2003. Sac3 is an mRNA export factor that localizes to cytoplasmic fibrils of nuclear pore complex. Mol. Biol. Cell. 14, 836-847 https://doi.org/10.1091/mbc.E02-08-0520
  11. 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
  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. Rodriguez-Navarro, S., T. Fischer, M. J. Luo, O. Antunez, S. Brettschneider, J. Lechner, J.E. Perez-Ortin, R. Reed, and E. Hurt. 2004. Sus1, a functional component of the SAGA histone acetylase complex and the nuclear pore-associated mRNA export machinery. Cell. 116, 75-86 https://doi.org/10.1016/S0092-8674(03)01025-0
  15. Rout, M.P., A.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
  16. 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
  17. Sommer, P. and U. Nehrbass. 2005. Quality control of messenger ribonucleoprotein particles in the nucleus and at the pore. Curr. Opin. Cell Biol. 17, 294-301 https://doi.org/10.1016/j.ceb.2005.04.007
  18. Strasser, K., S. Masuda, P. Mason, J. Pfannstiel, M. Oppizzi, S. Rodriguez-Navarro, A.G. Rondon, A. Aguilera, K. Struhl, R. Reed, and E. Hurt. 2002. TREX is a conserved complex coupling transcription with messenger RNA export. Nature. 417, 304-308 https://doi.org/10.1038/nature746
  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. Zenklusen, D., P. Vinciguerra, J.C. Wyss, and F. Stutz. 2002. Stable mRNP formation and export require cotranscriptional recruitment of the mRNA export factors Yra1p and Sub2p by Hpr1p. Mol. Cell. Biol. 22, 8241-8253 https://doi.org/10.1128/MCB.22.23.8241-8253.2002