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Studies on Intracellular Functions of the mas3 Gene in the Fission Yeast, Schizosaccharomyces pombe

분열형 효모에서 mas3 유전자의 세포내 기능 연구

  • Hwang Mi Ra (Division of Biological Sciences, Wonkwang university) ;
  • Cha Jae Young (Division of Biological Sciences, Wonkwang university) ;
  • Shin Sang Min (Division of Biological Sciences, Wonkwang university) ;
  • Park Jong Kun (Division of Biological Sciences, Wonkwang university)
  • 황미라 (원광대학교 자연대 생명과학부) ;
  • 차재영 (원광대학교 자연대 생명과학부) ;
  • 신상민 (원광대학교 자연대 생명과학부) ;
  • 박종군 (원광대학교 자연대 생명과학부)
  • Published : 2005.02.01

Abstract

The regulation of gene expression plays an important rolet in cell cycle controls. In this study, a novel $mas3^+$ (mitosis associated protein) gene, a homolog of human SMARCADl, was isolated and characterized from a fission yeast Schizosaccharomyces pombe. The overall homology between the helicase proteins of the two species is $87\%$. This DEAD/H box-containing molecule has seven highly conserved sequence regions that allow us to place it in the SNF2 family of the helicase superfamily. Knock-out cell of $mas3^+$ gene was constructed using kanMX6 as a selection marker. Survival of mas3 null mutant exposed to UV or MMS was similar to those of wild type cells. $mas3^+$ expression was lowest at $G_2$ and gradually increased. Cytokinesis of mas3 null mutant was abnormal at $26^{\circ}C\;and\;35^{\circ}C$ and a large number of multi-septate cells were produced. These results indicate that the $mas3^+$ is involved in cytokinesis and cell shape control.

세포주기 조절에서 유전자 발현의 조절은 매우 중요한 부분이다. 본 연구에서는 인간의 유전자인 SMARCADl과 상동성을 가지는 분열형 효모의 새로운 유전자 $mas3^+$를 분리하였다. 이 두 유전자는 $87\%$의 상동성을 보인다. $mas3^+$유전자는 DEAD/H box를 포함한 7개의 motif를 가지고 있어서 helicase superfamily 중에서도 SNF2 family에 속한다. kanMX6를 선별 표지로 이용하여 $mas3^+$유전자 결손 세포를 구성하였고 $mas3^+$ 유전자 결손 세포는 UV와 MMS처리 실험에서 정상의 세포와 생존율이 비슷하여 DNA상해회복과는 관련이 없음을 알 수 있었다. $mas3^+$ 유전자의 세포주기별 발현 양을 분석한 결과 $G_2$단계부터 점차적으로 발현양이 늘어났다. $mas3^+$결손 돌연변이를 $26^{\circ}C$$35^{\circ}C$에서 배양한 결과, 비정상적인 세포질 분열 과정으로 인해 다중 격막 세포의 빈도가 증가하였다. 이상의 결과들은 $mas3^+$유전자는 세포질 분열과 세포형태 형성에 연관되어 있음을 시사한다.

Keywords

References

  1. David, A., S. T. Guertin and M. C. Dannel. 2002. Cytokinesis in Eukaryotes. Micro. Mol. Bio Rev. 66, 155-178 https://doi.org/10.1128/MMBR.66.2.155-178.2002
  2. McInerny, C. J. 2004. Cell cycle-regulated transcription in fission yeast. Biochem. Soc. Trans. 32, 967-72 https://doi.org/10.1042/BST0320967
  3. Hirt, H., A. Pay, J. Gyorgyey, L. Bako, K. Nemeth, L. Bogre, R. J. Schweyen, E. Heberle-Bors and D. Dudits. 1991. Complementation of a yeast cell cycle mutant by an alfalfa cDNA encoding a protein kinase homologous to p34cdc2. Proc. Natl. Acad. Sci. USA. 88, 1636-40
  4. Jaspersen, S. L., B. J. Huneycutt, T. H. Giddings, K. A. Resing, N. G. Ahn and M. Winey. 2004. Cdc28/Cdk1 regulates spindle pole body duplication through phosphorylation of Spc42 and Mps1. Dev. Cell. 7, 263-74 https://doi.org/10.1016/j.devcel.2004.07.006
  5. Bartlett, R. and P. Nurse. 1990. Yeast as a model system for understanding the control of DNA replication in Eukaryotes. Bioessays. 12, 457-63 https://doi.org/10.1002/bies.950121002
  6. Russell, P. and P. Nurse. 1986. $cdc25^+$ functions as an inducer in the mitotic control of fission yeast. Cell. 45, 145-53 https://doi.org/10.1016/0092-8674(86)90546-5
  7. Krapp, A., E. Cano and V. Simanis. 2003. Mitotic hyperphosphorylation of the fission yeast SIN scaffold protein cdc11p is regulated by the protein kinase cdc7p. Curr. Biol. 13, 168-72 https://doi.org/10.1016/S0960-9822(02)01417-3
  8. Liu, J. L., P. Rigolet, S. X. Dou, P. Y. Wang and X. G. Xi. 2004. The zinc finger motif of Escherichia coli RecQ is implicated in both DNA binding and protein folding. J. Biol. Chem. 279, 42794-802 https://doi.org/10.1074/jbc.M405008200
  9. Lowe, J., A Sheerin, K. Jennert-Burston, D. Burton, E. L. Ostler, J. Bird, M. H. Green and R. G. Faragher. 2004. Camptothecin sensitivity in Werner syndrome fibroblasts as assessed by the COMET technique. Ann. N. Y. Acad. Sci. 1019, 256-9 https://doi.org/10.1196/annals.1297.042
  10. Garcia, P. L., Y. Liu, J. Jiricny, S. C. West and P. Janscak, 2004. Human RECQ5beta, a protein with DNA helicase and strand-annealing activities in a single polypeptide. EMBO J. 23, 2882-91 https://doi.org/10.1038/sj.emboj.7600301
  11. Miyajima A. 2002. Functional analysis of yeast homologue gene associated with human DNA helicase causative syndromes. Kokuritsu lyakuhin Shokuhin Eisei Kenkyusho Hokoku. 120, 53-74
  12. Van den Bosch, M., J. B. Zonneveld, K. Vreeken, F. A. de Vries. P. H. Lohman and A Pastink. 2002. Differential expression and requirements for Schizosaccharomyces pombe RAD52 homologs in DNA repair and recombination. Nucleic Acids Res. 30, 1316-24 https://doi.org/10.1093/nar/30.6.1316
  13. Cryer, D. R, R. Eccleshull and J. Marmur. 1975. in Methods cell Biology, Vol. 12, 39-44, Academic press
  14. Adra, C. N., J. L. Donato, R. Badovinac, F. Syed, R. Kheraj, H. Cai, C. Moran, M. T. Kolker, H. Turner, S. Weremowicz, T. Shirakawa, C. C. Morton, L. E. Schnipper and R. Drews. 2000. SMARCAD1, a novel human helicase family-defining member associated with genetic instability: cloning, expression, and mapping to 4q22-q23, a band rich in breakpoints and deletion mutants involved in several human diseases. Genomics. 69, 162-73 https://doi.org/10.1006/geno.2000.6281
  15. Catlett MG, Forsburg SL. 2003. Schizosaccharomyces pombe Rdh54 (TID1) acts with Rhp54 (RAD54) to repair meiotic double-strand breaks. Mol Biol Cell. 14, 4707-20 https://doi.org/10.1091/mbc.E03-05-0288
  16. Takeda, T., T. Kawate and F. Chang. 2004. Organization of a sterol-rich membrane domain by cdc15p during cytokinesis in fission yeast. Nat. Cell Biol. 6, 1142-4 https://doi.org/10.1038/ncb1189
  17. Krapp, A, S. Schmidt, E. Cano and V. Simanis. 2001. S. pombe cdc11p, together with sid4p, provides an anchor for septation initiation network proteins on the spindle pole body. Curr. Biol. 11, 1559-68 https://doi.org/10.1016/S0960-9822(01)00478-X
  18. Moreno, S., J. Hayles and P. Nurse. 1989. Regulation of p34cdc2 protein kinase during mitosis. Cell. 58, 361-72 https://doi.org/10.1016/0092-8674(89)90850-7
  19. Berlin, A., A. Paoletti and F. Chang. 2003. Mid2p stabilizes septin rings during cytokinesis in fission yeast. J. Cell Biol. 160, 1083-92 https://doi.org/10.1083/jcb.200212016
  20. Chang, L. and K. L. Gould. 2000. Sid4p is required to localize components of the septation initiation pathway to the spindle pole body in fission yeast. Proc. Natl. Acad. Sci. USA. 97. 5249-54