세포사멸 조절 단백질인 IEX-1의 새로운 결합단백질로서의 CATHEPSIN B의 발견
Identification of CATHEPSIN B as a Novel Binding Protein of the Cell Death Regulating Protein IEX-1
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선행연구에서 본 연구자는 IEX-1 단백질이 난소 암세포에서 세포사멸(apoptosis)을 유도하는 기능을 수행함을확인하였으나, 세포사멸과 세포생존의 여러 단계에서 어떠한 신호전달 체계로 IEX-1이 작용하는지는 정확히 알지 못하고있다. 따라서 IEX-1 단백질과 결합하는 새로운 단백질을 찾기 위해 yeast two-hybrid system을 이용하였다. 그 결과 IEX-1이 여러 다양한 인간 암 세포에서 세포사멸을 유도하는 CATHEPSIN B 단백질과 결합함을 밝혔다. 본 연구에서는 리소좀프로테아제의 일원인 CATHEPSIN B 단백질과 IEX-1 단백질의 결합을 면역침강법과 western blot 분석으로 확인하였다. 그러므로 이 결과들을 통해서 IEX-1과 CATHEPSIN B는 세포 내에서 서로의 기능에 영향을 미칠 것으로 예상되며, 세포사멸을 유도하는 IEX-1 단백질이 lysosome-mediated apoptotic pathway에 관여할 가능성을 시사한다.
Abstract
Previously, we identified that the overexpression of IEX-1 induces apoptosis in ovarian cancer cells. Herein we report a new binding partner of IEX-1, CATHEPSIN B, as a lysosomal enzyme which contributes to the various apoptotic signaling in tumor cells. To investigate how IEX-1 regulates cellular survival and death event, we performed yeast two-hybrid screening of rat ovarian cDNA library using IEX-1 as the bait and found CATHEPSIN B. In the present study,CATHEPSIN B and IEX-1 proteins were overexpressed in 293T cells and their specific association was determined by immunoprecipitation and immunoblot analysis. In addition, the endogenous interaction between CATHEPSIN B and IEX-1was confirmed in HeLa cells. The current finding of lysosomal CATHEPSIN B as the IEX-1-binding partner implies that IEX-1 may involve in lysosome-mediated apoptotic signaling pathways.
참고문헌 (27)
- Arlt A, "Expression of the NF-kappa B target gene IEX-1 does not prevent cell death but instead triggers apoptosis in Hela cells", Oncogene, 20, pp.69~76 (2001)
- Arlt A, "The early response gene IEX-1 attenuates NF-kappaB activation in 293 cells, a possible counter-regulatory process leading to enhanced cell death", Oncogene, 22, pp.3343~3351 (2003)
- Arlt A, "IEX-1 directly interferes with RelA", p65 dependent transactivation and regulation of apoptosis, 2008, pp.1783, 941~952
- Barrett AJ, "The possible role of neutrophil proteinases in damage to articular cartilage", Agents Actions, 43, pp.194~200 (1994)
- Charles CH, "Genomic structure, cDNA sequence, and expression of gly96, a growth factor-inducible immediate-early gene encoding a short-lived glycosylated protein", Oncogene, 8, pp.797~801 (1993)
- Felbor U, "Secreted cathepsin L generates endostatin from collagen XVIII", Embo J, 19, pp.1187~1194 (2000)
- Garcia J, "IEX-1: a new ERK substrate involved in both ERK survival activity and ERK activation", Embo J, 21, pp.5151~5163 (2002)
- Green DR, "Mitochondria and apoptosis", Science, 281, pp.1309~1312 (1998)
- Im HJ, "Characterization of a novel hexameric repeat DNA sequence in the promoter of the immediate early gene, IEX-1, that mediates 1alpha,25-dihydroxyvitamin D(3)- associated IEX-1 gene repression", Oncogene, 21, pp.3706~3714 (2002)
- 김재홍, "KGN(난소과립세포)에서 BCL2L10 단백질의 세포사멸 유도 기능 연구", 발생과 생식, 15(2), pp.113~120 (2011), 한국발생생물학회
- Kobayashi T, "Regulation of a novel immediate early response gene, IEX-1, in keratinocytes by 1alpha, 25-dihydroxyvitamin D3", Biochem Biophys Res Commun, 251, pp.868~873 (1998)
- Koblinski JE, "Unraveling the role of proteases in cancer", Clin Chim Acta, 291, pp.113~135 (2000)
- Kruse ML, "Immediate early gene X1 is organized in subnuclear structures and partially co-localizes with promyelocytic leukemia protein in HeLa cells", J Biol Chem, 280, pp.24849~24856 (2005)
- Kumar R, "Immediate early gene X-1 interacts with proteins that modulate apoptosis", Biochem Biophys Res Commun, 323, pp.1293~1298 (2004)
- Li W, "3-Aminopropanal, formed during cerebral ischaemia, is a potent lysosomotropic neurotoxin", Biochem J, 371, pp.429~436 (2003)
- Li W, "Induction of cell death by the lysosomotropic detergent MSDH", FEBS Lett, 470, pp.35~39 (2000)
- Manna SK, "Overexpression of manganese superoxide dismutase suppresses tumor necrosis factor-induced apoptosis and activation of nuclear transcription factorkappaB and activated protein-1", J Biol Chem, 273, pp.13245~13254 (1998)
- Osawa Y, "Expression of the NF-kappa B target gene X-ray-inducible immediate early response factor-1 short enhances TNF-alphainduced hepatocyte apoptosis by inhibiting Akt activation", J Immunol, 170, pp.4053~4060 (2003)
- Schilling D, "IEX-1, an immediate early gene, increases the rate of apoptosis in keratinocytes", Oncogene, 20, pp.7992~7997 (2001)
- Shen L, "Distinct domains for anti- and pro-apoptotic activities of IEX-1", J Biol Chem, 281, pp.15304~15311 (2006)
- Stoka V, "Lysosomal protease pathways to apoptosis. Cleavage of bid, not pro-caspases, is the most likely route", J Biol Chem, 276, pp.3149~3157 (2001), is the most likely route. J Biol Chem 276 : 3149-3157.
- Tait DL, "Ovarian cancer BRCA1 gene therapy: Phase I and II trial differences in immune response and vector stability", Clin Cancer Res, 5, pp.1708~1714 (1999)
- Turk B, "Apoptotic pathways: involvement of lysosomal proteases", Biol Chem, 383, pp.1035~1044 (2002)
- Vancompernolle K, "Atractyloside-induced release of cathepsin B, a protease with caspase-processing activity", FEBS Lett, 438, pp.150~158 (1998)
- Yoon S, "IEX-1-induced cell death requires BIM and is modulated by MCL-1", Biochem Biophys Res Commun, 382, pp.400~404 (2009)
- 윤성민, "난소암 세포에서 IEX-1과 MCL-1 단백질들의 세포사멸 기능에 관한 상호작용", 발생과 생식, 14(2), pp.83~89 (2010), 한국발생생물학회
- Zhang Y, "Development of T-cell lymphomas in Emu-IEX-1 mice", Oncogene, 22, pp.6845~6851 (2003)