Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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APEX-1 Regulates Cell Proliferation through GDNF/GFRα1 Signaling

APEX-1은 GDNF/GFRα1 시그널을 통해 세포증식을 조절한다

  • Received : 2013.06.28
  • Accepted : 2013.10.08
  • Published : 2013.10.30
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Abstract

Human apurinic/apyrimidinic endonuclease (APEX-1) is a multifunctional protein that is capable of repairing abasic sites and single-strand breaks in damaged DNA. In addition, it serves as a redox-modifying factor for a number of transcription factors. Identifying the transcriptional targets of APEX-1 is essential for understanding how it affects various cellular outcomes. Expression array analysis was used to identify glial cell-derived neurotropic factor receptor ${\alpha}1$ ($GFR{\alpha}1$), which is an encoding receptor for the glial cell-derived neurotropic factor (GDNF) family, the expression of which is induced by APEX-1. A target of GDNF/$GFR{\alpha}$ signaling, c-Src (Tyr418) was strongly phosphorylated by GNDF in the APEX-1 expressing cells. Moreover, GDNF initiated cell proliferation, measured by counting the number of cells, in the APEX-1 expressing cells. Importantly, the down-regulation of APEX-1 by siRNA caused a marked reduction in the $GFR{\alpha}1$ expression level, and it reduced the ability of GDNF to phosphorylate c-Src (Tyr418) and stimulate cell proliferation. These results demonstrate an association between APEX-1 and GDNF/$GFR{\alpha}$ signaling and suggest a potential molecular mechanism for the involvement of APEX-1 in cell survival and proliferation.

APEX-1 (인간 apyrimidinic/apurinic 효소)은 염기성 사이트 및 DNA단일 가닥 결손으로 손상된 DNA을 복구 할 수 있는 다기능 단백질이다. 또한 APEX-1은 많은 전사 인자들의 redox-modifying factor (산화 환원 수정 요소)로서의 역할을 한다고 알려져 있다. 이런 APEX-1의 전사 타겟을 동정하는 것은 APEX-1의 다양한 세포 내 작용 메커니즘을 이해하는데 필수적이다. 따라서 이 논문에서는 먼저 Expression array analysis를 통해 glial cell-derived neurotropic factor receptor ${\alpha}1$ ($GFR{\alpha}1$)을 동정하였다. $GFR{\alpha}1$은 glial cell-derived neurotropic factor (GDNF) family 수용체이며 APEX-1에 의해 발현이 증가된다. APEX-1이 과발현된 세포에서 GDNF처리에 의해 GDNF/$GFR{\alpha}1$ 시그널 타겟인 c-Src가 Tyr418잔기에서 인산화 됨을 관찰하였다. 또한 APEX-1이 과발현된 세포에 GDNF처리하면, 세포증식이 증가함을 보았다. 반면, APEX-1 발현을 siRNA을 이용하여 감소시키면 $GFR{\alpha}1$ 발현과 GDNF에 의한 c-Src 인산화 및 세포증식이 감소함을 확인하였다. 이상의 결과는 APEX-1은 GDNF/$GFR{\alpha}1$ 시그널을 통해 세포 생존과 증식을 조절함을 증명하였다. 따라서 본 연구를 통해 APEX-1의 세포 증식을 조절하는 새로운 기전을 규명하였다.

Keywords

References

  1. Airaksinen, M. S. and Saarma, M. 2002. The GDNF family: signalling, biological functions and therapeutic value. Nat Rev Neurosci 3, 383-394. https://doi.org/10.1038/nrn812
  2. Airaksinen, M. S., Titievsky, A. and Saarma, M. 1999. GDNF family neurotrophic factor signaling: four masters, one servant? Mol Cell Neurosci 13, 313-325. https://doi.org/10.1006/mcne.1999.0754
  3. Bobola, M. S., Blank, A., Berger, M. S., Stevens, B. A. and Silber, J. R. 2001. Apurinic/apyrimidinic endonuclease activity is elevated in human adult gliomas. Clin Cancer Res 7, 3510-3518.
  4. Cacalano, G., Farinas, I., Wang, L. C., Hagler, K., Forgie, A., Moore, M., Armanini, M., Phillips, H., Ryan, A. M., Reichardt, L. F., Hynes, M., Davies, A. and Rosenthal, A. 1998. GFRalpha1 is an essential receptor component for GDNF in the developing nervous system and kidney. Neuron 21, 53-62. https://doi.org/10.1016/S0896-6273(00)80514-0
  5. Evans, A. R., Limp-Foster, M. and Kelley, M. R. 2000. Going APE over ref-1. Mutat Res 461, 83-108. https://doi.org/10.1016/S0921-8777(00)00046-X
  6. Fritz, G., Grosch, S., Tomicic, M. and Kaina, B. 2003. APE/Ref-1 and the mammalian response to genotoxic stress. Toxicology 193, 67-78. https://doi.org/10.1016/S0300-483X(03)00290-7
  7. Grosch, S., Fritz, G. and Kaina, B. 1998. Apurinic endonuclease (Ref-1) is induced in mammalian cells by oxidative stress and involved in clastogenic adaptation. Cancer Res 58, 4410-4416.
  8. Iwahashi, N., Nagasaka, T., Tezel, G., Iwashita, T., Asai, N., Murakumo, Y., Kiuchi, K., Sakata, K., Nimura, Y. and Takahashi, M. 2002. Expression of glial cell line-derived neurotrophic factor correlates with perineural invasion of bile duct carcinoma. Cancer 94, 167-174. https://doi.org/10.1002/cncr.10169
  9. Jing, S., Wen, D., Yu, Y., Holst, P. L., Luo, Y., Fang, M., Tamir, R., Antonio, L., Hu, Z., Cupples, R., Louis, J. C., Hu, S., Altrock, B. W. and Fox, G. M. 1996. GDNF-induced activation of the ret protein tyrosine kinase is mediated by GDNFR-alpha, a novel receptor for GDNF. Cell 85, 1113-1124. https://doi.org/10.1016/S0092-8674(00)81311-2
  10. Kelley, M. R., Cheng, L., Foster, R., Tritt, R., Jiang, J., Broshears, J. and Koch, M. 2001. Elevated and altered expression of the multifunctional DNA base excision repair and redox enzyme Ape1/ref-1 in prostate cancer. Clin Cancer Res 7, 824-830.
  11. Klein, R. D., Sherman, D., Ho, W. H., Stone, D., Bennett, G. L., Moffat, B., Vandlen, R., Simmons, L., Gu, Q., Hongo, J. A., Devaux, B., Poulsen, K., Armanini, M., Nozaki, C., Asai, N., Goddard, A., Phillips, H., Henderson, C. E., Takahashi, M. and Rosenthal, A. 1997. A GPI-linked protein that interacts with Ret to form a candidate neurturin receptor. Nature 387, 717-721. https://doi.org/10.1038/42722
  12. Lin, L. F., Doherty, D. H., Lile, J. D., Bektesh, S. and Collins, F. 1993. GDNF: a glial cell line-derived neurotrophic factor for midbrain dopaminergic neurons. Science 260, 1130-1132. https://doi.org/10.1126/science.8493557
  13. Meng, X., Lindahl, M., Hyvonen, M.E., Parvinen, M., de Rooij, D. G., Hess, M. W., Raatikainen-Ahokas, A., Sainio, K., Rauvala, H., Lakso, M., Pichel, J. G., Westphal, H., Saarma, M. and Sariola, H. 2000. Regulation of cell fate decision of undifferentiated spermatogonia by GDNF. Science 287, 1489-1493. https://doi.org/10.1126/science.287.5457.1489
  14. Moore, D. H., Michael, H., Tritt, R., Parsons, S. H. and Kelley, M. R. 2000. Alterations in the expression of the DNA repair/redox enzyme APE/ref-1 in epithelial ovarian cancers. Clin Cancer Res 6, 602-609.
  15. Moore, M. W., Klein, R. D., Farinas, I., Sauer, H., Armanini, M., Phillips, H., Reichardt, L. F., Ryan, A. M., Carver- Moore, K. and Rosenthal, A. 1996. Renal and neuronal abnormalities in mice lacking GDNF. Nature 382, 76-79. https://doi.org/10.1038/382076a0
  16. Pichel, J. G., Shen, L., Sheng, H. Z., Granholm, A. C., Drago, J., Grinberg, A., Lee, E. J., Huang, S. P., Saarma, M., Hoffer, B. J. Sariola, H. and Westphal, H. 1996. Defects in enteric innervation and kidney development in mice lacking GDNF. Nature 382, 73-76. https://doi.org/10.1038/382073a0
  17. Popsueva, A., Poteryaev, D., Arighi, E., Meng, X., Angers-Loustau, A., Kaplan, D., Saarma, M. and Sariola, H. 2003. GDNF promotes tubulogenesis of GFRalpha1-expressing MDCK cells by Src-mediated phosphorylation of Met receptor tyrosine kinase. J Cell Biol 161, 119-129. https://doi.org/10.1083/jcb.200212174
  18. Sanchez, M. P., Silos-Santiago, I., Frisen, J., He, B., Lira, S. A. and Barbacid, M. 1996. Renal agenesis and the absence of enteric neurons in mice lacking GDNF. Nature 382, 70-73. https://doi.org/10.1038/382070a0
  19. Veit, C., Genze, F., Menke, A., Hoeffert, S., Gress, T. M., Gierschik, P. and Giehl, K. 2004. Activation of phosphatidylinositol 3-kinase and extracellular signal-regulated kinase is required for glial cell line-derived neurotrophic factor-induced migration and invasion of pancreatic carcinoma cells. Cancer Res 64, 5291-5300. https://doi.org/10.1158/0008-5472.CAN-04-1112
  20. Walker, L. J., Craig, R. B., Harris, A. L. and Hickson, I. D. 1994. A role for the human DNA repair enzyme HAP1 in cellular protection against DNA damaging agents and hypoxic stress. Nucleic Acids Res 22, 4884-4889. https://doi.org/10.1093/nar/22.23.4884
  21. Wilson, T. M., Rivkees, S. A., Deutsch, W. A. and Kelley, M. R. 1996. Differential expression of the apurinic / apyrimidinic endonuclease (APE/ref-1) multifunctional DNA base excision repair gene during fetal development and in adult rat brain and testis. Mutat Res 362, 237-248. https://doi.org/10.1016/0921-8777(95)00053-4
  22. Xanthoudakis, S. and Curran, T. 1992. Identification and characterization of Ref-1, a nuclear protein that facilitates AP-1 DNA-binding activity. EMBO J 11, 653-665.
  23. Xanthoudakis, S., Smeyne, R. J., Wallace, J. D. and Curran, T. 1996. The redox/DNA repair protein, Ref-1, is essential for early embryonic development in mice. Proc Natl Acad Sci USA 93, 8919-8923. https://doi.org/10.1073/pnas.93.17.8919
  24. Xu, Y., Moore, D. H., Broshears, J., Liu, L., Wilson, T. M. and Kelley, M. R. 1997. The apurinic/apyrimidinic endonuclease (APE/ref-1) DNA repair enzyme is elevated in premalignant and malignant cervical cancer. Anticancer Res 17, 3713-3719.