Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Regulation of the Drosophila Fascin by Raf Signaling

Raf 신호에 의한 초파리 fascin의 조절

  • Pyo, Jung-Hoon (Department of Molecular Biology, College of Natural Science, Pusan National University) ;
  • Choi, Na-Hyun (Department of Molecular Biology, College of Natural Science, Pusan National University) ;
  • Lee, Shin-Hae (Department of Molecular Biology, College of Natural Science, Pusan National University) ;
  • Kim, Young-Shin (Department of Molecular Biology, College of Natural Science, Pusan National University) ;
  • Yoo, Mi-Ae (Department of Molecular Biology, College of Natural Science, Pusan National University)
  • 표정훈 (부산대학교 자연과학대학 분자생물학과) ;
  • 최나현 (부산대학교 자연과학대학 분자생물학과) ;
  • 이신해 (부산대학교 자연과학대학 분자생물학과) ;
  • 김영신 (부산대학교 자연과학대학 분자생물학과) ;
  • 유미애 (부산대학교 자연과학대학 분자생물학과)
  • Published : 2009.07.30
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Abstract

Fascin, a cytoskeleton actin binding protein, functions in cell adhesion and cell migration. Fascin is also known as a candidate biomarker for various cancers, however, regulatory mechanisms of fascin expression remains little understood. In this study, we found an abnormal bristle phenotype, which is similar to that of the Drosophila fascin mutant, in Drafmutant flies. Hence, we investigated whether fascin expression is regulated by Raf signaling. RT-PCR and Western blot analysis showed that Drosophila fascin expression was down-regulated in Draf mutant flies and the level was increased in larvae expressing the oncogenic form of Draf (Draf$^{got}$) under the GAL4-UAS system. Immunostaining analysis showed increased fascin in the hemocytes over-expressing Draf$^{got}$. Our results indicate that fascin expression is regulated by Raf signaling and suggest that Raf signaling may play an important role in the actin cytoskeleton-associated developmental process and tumor progression via regulation of fascin gene.

Fascin은 액틴결합 단백질로 형성을 포함한 많은 발생과정에 있어서 중요한 역할을 한다. Fascin은 암세포에 대한 생체표지인자로도 잘 알려져 있다. 그러나 이러한 fascin 유전자의 발현조절기전은 현재까지 잘 알려져있다. 그러나 이러한 fascin 유전자의 발현조절기전은 현재까지 잘 알려져 있지 않다. 본 연구에서는 Raf 돌연변이 초파리에서 이미 보고 되어있는 초파리 fascin 돌연변이 초파리의 휘어진 등털 표현형을 관찰함으로써 초파리 fascin 유전자의 발현이 Raf신호체계에 의해 조절되는 가를 조사하였다. RT-PCR과 Western blot의 실험 방법으로 Raf 유전자 돌연변이 초파리에서 fascin의 발현이 감소되어 있는 것을 확인하였으며, GAL4-UAS계로 Raf를 과발현시킨 초파리에서 fascin 발현이 증가하는 것을 확인할 수 있었다. 또한 세포의 증식과 이동 연구모델계로 잘 알려져 있는 초파리의 혈액세포에서 이러한 조절기전을 확인하였다. 이러한 결과들은 fascin유전자의 발현이 Raf 신호체계에 의해 조절된다는 것을 나타낸다.

Keywords

References

  1. Adams, J. C. 2004. Roles of fascin in cell adhesion and motility. Curr. Opin. Cell Biol. 16, 590-596 https://doi.org/10.1016/j.ceb.2004.07.009
  2. Adams, J. C., J. D. Clelland, G. D. Collett, F. Matsumura, S. Yamashir, and L. Zhang. 1999. Cell-matrix adhesions differentially regulate fascin phosphorylation. Mol. Biol. Cell 10, 4177-4190 https://doi.org/10.1093/emboj/cdg521
  3. Asha, H., I. Nagy, G. Kovacs, D. Stetson, I. Ando, and C. R. Dearolf. 2003. Analysis of Ras-induced overproliferation in Drosophila hemocytes. Genetics 163, 203-215
  4. Brand, A. H. and N. Perrimon. 1993. Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development. 118, 401-415
  5. Bros, M., X. L. Ross, A. Pautz, A. B. Reske-Kunz, and R. Ross. 2003. The human fascin gene promoter is highly active in mature dendritic cells due to a stage-specific enhancer. J. Immunol. 171, 1825-1834 https://doi.org/10.4049/jimmunol.171.4.1825
  6. Bryan, J., R. Edwards, P. Matsudaira, J. Otto, and J. Wulfkuhle. 1993. Fascin, an echinoid actin-bundling protein, is a homolog of the Drosophila singed gene product. Proc. Natl. Acad. Sci. U. S. A. 90, 9115-9119 https://doi.org/10.1073/pnas.90.19.9115
  7. Buda, A. and M. Pignatelli. 2004. Cytoskeletal network in colon cancer: from genes to clinical application. Int. J. Biochem. Cell Biol. 36, 759-765 https://doi.org/10.1016/j.biocel.2003.09.004
  8. Cant, K., B. A. Knowles, M. S. Mooseker, and L. Cooley. 1994. Drosophila singed, a fascin homolog, is required for actin bundle formation during oogenesis and bristle extension. J. Cell Biol. 125, 369-380 https://doi.org/10.1083/jcb.125.2.369
  9. De Arcangelis, A., E. Georges-Labouesse, and J. C. Adams. 2004. Expression of fascin-1, the gene encoding the actin-bundling protein fascin-1, during mouse embryogenesis. Gene Expr. Patterns 4, 637-643 https://doi.org/10.1016/j.modgep.2004.04.012
  10. Guan, Y., P. L. Woo, N. M. Rubenstein, and G. L. Firestone. 2002. Transforming growth factor-alpha abrogates the glucocorticoid stimulation of tight junction formation and reverses the steroid-induced down-regulation of fascin in rat mammary epithelial tumor cells by a Ras-dependent pathway. Exp. Cell Res. 273, 1-11 https://doi.org/10.1006/excr.2001.5415
  11. Hashimoto, Y., M. Parsons, and J. C. Adams. 2007. Dual actin-bundling and protein kinase C-binding activities of fascin regulate carcinoma cell migration downstream of Rac and contribute to metastasis. Mol. Biol. Cell 18, 4591-4602 https://doi.org/10.1091/mbc.E07-02-0157
  12. Helfman, D. M. and G. Pawlak. 2005. Myosin light chain kinase and acto myosin contractility modulate activation of the ERK cascade downstream of oncogenic Ras. J. Cell Biochem. 95, 1069-1080 https://doi.org/10.1002/jcb.20498
  13. Howe, A. K., A. E. Aplin, and R. L. Juliano. 2002. Anchorage-dependent ERK signaling--mechanisms and consequences. Curr. Opin. Genet. Dev. 12, 30-35 https://doi.org/10.1016/S0959-437X(01)00260-X
  14. Kolch, W. 2005. Coordinating ERK/MAPK signalling through scaffolds and inhibitors. Nat. Rev. Mol. Cell Biol. 6, 827-837 https://doi.org/10.1038/nrm1743
  15. Kureishy, N., V. Sapountzi, S. Prag, N. Anilkumar, and J. C. Adams. 2002. Fascins, and their roles in cell structure and function. Bioessays. 24, 350-361 https://doi.org/10.1002/bies.10070
  16. Melnick, M. B., L. A. Perkins, M. Lee, L. Ambrosio, and N. Perrimon. 1993. Developmental and molecular characterization of mutations in the Drosophila-raf serine/threonine protein kinase. Development 118, 127-138
  17. Otto, J. J., R. E. Kane, and J. Bryan. 1979. Formation of filopodia in coelomocytes: localization of fascin, a 58,000 dalton actin cross-linking protein. Cell 17, 285-293 https://doi.org/10.1016/0092-8674(79)90154-5
  18. Paterson, J. and K. O'Hare. 1991. Structure and transcription of the singed locus of Drosophila melanogaster. Genetics 129, 1073-1084
  19. Patwari, P. and R. T. Lee. 2008. Mechanical control of tissue morphogenesis. Circ. Res. 103, 234-243 https://doi.org/10.1161/CIRCRESAHA.108.175331
  20. Phelps, C. B. and A. H. Brand. 1998. Ectopic gene expression in Drosophila using GAL4 system. Methods 14, 367-379 https://doi.org/10.1006/meth.1998.0592
  21. Ramos, J. W. 2008. The regulation of extracellular signalregulated kinase (ERK) in mammalian cells. Int. J. Biochem. Cell Biol. 40, 2707-2719 https://doi.org/10.1016/j.biocel.2008.04.009
  22. Xie, J., J. Qian, J. Yang, S. Wang, M. E. Freeman 3rd, and Q. Yi. 2005. Critical roles of Raf/MEK/ERK and PI3K/AKT signaling and inactivation of p38 MAP kinase in the differentiation and survival of monocyte-derived immature dendritic cells. Exp. Hematol. 33, 564-572 https://doi.org/10.1016/j.exphem.2005.03.001