Analysis and characterization of sumoylation substrates in anti-cancer drugs-related apoptosis

• Project/Area Number: 16590966
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Hematology
• Research Institution: JAPANESE FOUNDATION FOR CANCER RESEARCH
• Principal Investigator: TERUI Yasuhito JAPANESE FOUNDATION FOR CANCER RESEARCH, RESEARCHER, 癌化学療法センター臨床部, 副部長 (10285786)
• Project Period (FY): 2004 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥3,100,000 (Direct Cost: ¥3,100,000); Fiscal Year 2005: ¥1,200,000 (Direct Cost: ¥1,200,000); Fiscal Year 2004: ¥1,900,000 (Direct Cost: ¥1,900,000)
• Keywords: SUMO-1 / anti-cancer drugs / SUMO foci / apoptosis / GFP-SUMO-1 / K562 / molecular target / SUMO修飾 / NFAT1 / 転写活性 / 核内保持

Research Abstract

The nuclear import of NFAT transcription factors is critical for regulating its activity. Here we demonstrate that the sumoylation of NFAT1 defines a novel mechanism of the nuclear anchorage and transcriptional activation downstream from the known mechanism of calcineurin-mediated dephosphorylation and nuclear import. We show that K684 and K897 of NFAT1 can be sumoylated. The sumoylation at K684 is required for NFAT1 transcriptional activity and subsequent sumoylation of K897 whereas the sumoylation of K897 is only required for nuclear anchorage. Since K897 of NFAT1 is not conserved among other members of the NFAT family, we propose that sumoylation of K897 may provide a mechanism for NFAT1 isotype specific regulation of nuclear anchorage and transcriptional activation. Furthermore, we found that the treatment of both ionomycin and PMA ensured an efficient nuclear anchorage with the recruitment of NFAT1 into the SUMO-1 bodies, while the treatment of ionomycin alone induces the nuclear translocation of NFAT1 but not the recruitment into the SUMO-1 bodies. Our results suggest that the recruitment of NFAT1 into SUMO-1 bodies may be required for the progressive transcriptional activity of NFAT1 upon co-stimulation of ionomycin and PMA, whereas anergic transcription stimulated by ionomycin alone may occur without being recruited into the SUMO-1 bodies. As exploring the relationship among sumoylation, anti-tumor reagents, and apoptosis, intracellular GFP-SUMO-1 showed three types of subcellular distribution ; (1)GFP-SUMO-1 located homogeneously in nuclei by treatment with TPA ; (2)GFP-SUMO-1 formed foci in nuclei by treatment with doxorubicin, etoposide (VP-16), mitoxantrone (MIT), imatinib, and IFN-a ; (3)Both the first type and the second type were seen by treatment with arabinoside (AraC), fludarabine, cisplatin (CDDP), and methotrexate (MTX). When K562/GFP-SUMO-1 cells were treated with TPA plus MIT, GFP-SUMO-1 foci became larger in nuclei, and apoptosis was induced more than with MIT alone.

Research Products

• [Journal Article] Blockade of bulky lymphoma-associated CD55 expression by RNA interference overcomes resistance to CDC with rituximab. (2006)
  • Author(s): Terui Y, Sakurai T, Mishima Y, Mishima Y, Sugimura N, Sasaoka C, Kojima K, Yokoyama M, Mizunuma N, Takahashi S, Ito Y, Hatake K
  • Journal Title: Cancer Science 97(1) Pages: 72-79
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Differential Regulation of Eotaxin-1/CCL11 and Eotaxin-3/CCL26 Production by the TNF-a and IL-4 stimulated Human Lung Fibroblast. (2006)
  • Author(s): Rokudai A, Terui Y, Kuniyoshi R, Mishima Y, Mishima Y, Aizu-Yokota Y, Sonoda Y, Kasahara T, Hatake K
  • Journal Title: Biol. Pharm. Bull. 29(In press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Blockade of bulky lymphoma-associated CD55 expression by RNA interference overcomes resistance to CDC with rituximab. (2006)
  • Author(s): Terui Y, Sakurai T, Mishima Y, Mishima Y, Sugimura N, Sasaoka C, Kojima K, Yokoyama M, Mizunuma N, Takahashi S, Ito Y, Hatake K
  • Journal Title: Cancer Science. 97 Pages: 72-79
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Differential Regulation of Eotaxin-1/CCL11 and Eotaxin-3/CCL26 Production by the TNF-a and IL-4 stimulated Human Lung Fibroblast. (2006)
  • Author(s): Rokudai A, Terui Y, Kuniyoshi R, Mishima Y, Mishima Y, Aizu-Yokota Y, Sonoda Y, Kasahara T, Hatake K
  • Journal Title: Biol.Pharm.Bull. 29(in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Differential Regulation of Eotaxin-1/CCL11 and Eotaxin-3/CCL26 Production by the TNF-a and IL-4 stimulated Human Lung Fibroblast. (2006)
  • Author(s): Rokudai A, Terui Y, Kuniyoshi R, Mishima Y, Mishima Y, Aizu-Yokota Y, Sonoda Y, Kasahara T, Hatake K
  • Journal Title: Biol.Pharm.Bull. 29(In press)
• [Journal Article] Element array by scanning X-ray fluorescence microscopy after cis-diamminedichloro-platinum (II) treatment. (2005)
  • Author(s): Shimura M, Saito A, Matsuyama S, Sakuma T, Terui Y, Ueno K, Yumoto H, Yamauchi K, Yamamura K, Mimura H, Sano Y, Yabashi M, Tamasaku K, Nishio K, Nishino Y, Endo K, Hatake K, Mori Y, Ishizaka Y, Ishikawa T:
  • Journal Title: Cancer Res 65 Pages: 4998-5002
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Element array by scanning X-ray fluorescence microscopy after cis-diamminedichloro-platinum (II) treatment. (2005)
  • Author(s): Shimura M, Saito A, Matsuyama S, Sakuma T, Terui Y, Ueno K, Yumoto H, Yamauchi K, Yamamura K, Mimura H, Sano Y, Yabashi M, Tamasaku K, Nishio K, Nishino Y, Endo K, Hatake K, Mori Y, Ishizaka Y, Ishikawa T
  • Journal Title: Cancer Res. 65 Pages: 4998-5002
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Element array by scanning X-ray fluorescence microscopy after cis-diamminedichloro-platinum (II) treatment. (2005)
  • Author(s): Shimura M, Saito A, Matsuyama S, Sakuma T, Terui Y, Ueno K, Yumoto H, yamauchi K, Yamamura K, Mimura H, Sano Y, Yabashi M, Tamasaku K, Nishio K, Nishino Y, Endo K, Hatake K, Mori Y, Ishizaki Y, Ishikawa T
  • Journal Title: Cancer Res 65 Pages: 4998-5002
• [Journal Article] Dual role of sumoylation in the nuclear localization and transcriptional activation of NFAT1 (2004)
  • Author(s): Terui Y. et al.
  • Journal Title: Journal of Biological Chemistry 279 Pages: 28257-28265
• [Journal Article] Combination chemotherapy, CADO-CVP, with radiotherapy and stem cell support can be beneficial for adolescent and adult estheisoneuroblastoma (2004)
  • Author(s): Mishima Y, Nagasaki E, Terui Y, et al.
  • Journal Title: Cancer. 101 Pages: 1437-1444