Functional analyses of chromatin structure changing-related factors in higher eukaryotic cells by gene targeting techniques

• Project/Area Number: 19310127
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 基礎ゲノム科学
• Research Institution: University of Miyazaki
• Principal Investigator: NAKAYAMA Tatsuo University of Miyazaki, 理事 (60031712)
• Co-Investigator(Kenkyū-buntansha): TAKAMI Yasunari 宮崎大学, 医学部, 准教授 (80236356) ; KIKUCHI Hidehiko 宮崎大学, 医学部, 助教 (10301384)
• Project Period (FY): 2007 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥20,410,000 (Direct Cost: ¥15,700,000、Indirect Cost: ¥4,710,000); Fiscal Year 2009: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000); Fiscal Year 2008: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000); Fiscal Year 2007: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000)
• Keywords: ヒストン / クロマチン / ヌクレオソーム / ジーンノックアウト法 / アポトーシス / 転写制御 / クロマチン構造 / ヒストンシャペロン / DNA複製 / 細胞周期 / チェックポイント / DNA障害 / DT40 / ジーンノックアスト法

Research Abstract

To clarify individual roles of chromatin structure changing-related factors in higher eukaryotic cells, we generated systematically a number of chicken DT40 mutant cell lines, respectively, lacking each of appropriate genes. By analyses of these mutant cells, we obtained important results as follows. (1) CAF-1-dependent nucleosome assembly is essential in DNA replication and cell proliferation via its interaction with PCNA. (2) HDAC2 regulates indirectly gene expressions of IgM H- and ・L chains via opposite controls of gene expressions of EBF1, Pax5, Aiolos plus Ikaros, and E2A. (3) GCN5 and BCR signaling regulate cooperatively pre-mature B cell apoptosis through both increases of caspase-3 and caspase-8 activities and decreases of ICAD and IAP2.

Research Products

• [Journal Article] GCN5 regulates the activation of PI3K/Akt survival pathway in B cells exposed to oxidative stress via controlling gene expressions of Syk and Btk. (2011)
  • Author(s): Kikuchi, H., Kuribayashi, F., Takami, Y., Imajoh-Ohmi, S., Nakayama, T.
  • Journal Title: Biochem. Biophys. Res. Commun. 405 Pages: 657-661
  • Peer Reviewed
• [Journal Article] GCN5 regulates the superoxide-generating system in Leukocytes via controlling gp91-phox gene expression. (2011)
  • Author(s): Kikuchi, H., Kuribayashi, F., Kiwaki, N., Takami, Y., Nakayama, T.
  • Journal Title: J. Immunology 186 Pages: 3015-3022
  • Peer Reviewed
• [Journal Article] Improved applications of the tetracycline-regulated gene depletion system. (2010)
  • Author(s): Nishijima, H., Takami, Y., Nakayama, T., Adachi, N., Shibahara, K.
  • Journal Title: BioScience Trends 3 Pages: 161-167
• [Journal Article] Histone H1 null vertebrate cells exhibit altered nucleosome architecture. (2010)
  • Author(s): Hashimoto, H., Takami, Y., Sonoda, E., Iwasaki, T., Iwano, H., Tachibana, M., Takeda, S., Nakayama, T., Kimura, H., Shinkai, Y.
  • Journal Title: Nucleic Acids Res. 38 Pages: 3533-3545
  • Peer Reviewed
• [Journal Article] Curcumin dramatically enhances retinoic acid-induced superoxide generating activity via accumulation of p47-phox and p67-phox proteins in U937 cells. (2010)
  • Author(s): Kikuchi, H., Kuribayashi, F., Kiwaki, N., Nakayama, T.
  • Journal Title: Biochem. Biophys. Res. Commun. 395 Pages: 61-65
  • Peer Reviewed
• [Journal Article] Histone H1 null vertebrate cells exhibit altered nucleosome architecture (2010)
  • Author(s): Hashimoto H.
  • Journal Title: Nucleic Acids Res. Volume: 38 Pages: 3533-3545
  • Peer Reviewed
• [Journal Article] E2A participates in a fine control of pre-mature B cell apoptosis mediated by B cell receptor signaling via transcription regulations of survivin, IAP2 and caspase-8 genes. (2009)
  • Author(s): Toyonaga, K., Kikuchi, H., Yamashita, K., Nakayama, M., Chijiiwa, K., Nakayama, T.
  • Journal Title: FEBS J. 276/5 Pages: 1418-1428
  • Peer Reviewed
• [Journal Article] Lacking of Aiolos accelerates pre-mature B cell apoptosis mediated by BCR signaling through elevation in cytochrome c release. (2009)
  • Author(s): Kikuchi, H., Yamashita, K., Nakayama, M., Toyonaga, K., Tsuneyoshi, I., Takasaki, M., Nakayama, T.
  • Journal Title: BBA-Molecular Cell Research 1793 Pages: 1304-1314
  • Peer Reviewed
• [Journal Article] E2A participates in a fine control of pre-mature B-cell apoptosis mediated by B-cell receptor signaling via transcriptional regulation of survivin, IAP2 and caspase-8 (2009)
  • Author(s): Toyonaga K.
  • Journal Title: FEBS J. Volume: 276 Pages: 1418-1428
  • Peer Reviewed
• [Journal Article] Lacking of Aiolos accelerates pre-mature B cell apoptosis mediated by BCR signaling through elevation in cytochrome c release (2009)
  • Author(s): Kikuchi H.
  • Journal Title: Biochim Biophys Acta. Volume: 1793 Pages: 1304-1314
  • Peer Reviewed
• [Journal Article] Improved applications of the tetracycline-regulated gene depletion system (2009)
  • Author(s): Nishijima H.
  • Journal Title: BioScience Trends Volume: 3 Pages: 161-167
• [Journal Article] E2A participates in a fine control of pre-mature B cell apoptosis mediated by B cell receptorsignaling via transcription regulations of survivin, IAP2 and caspase-8 genes. (2009)
  • Author(s): *Toyonaga, K., *Kikuchi, H., Yamashita, K., Nakayama, M., Chijiiwa, K., Nakayama, T.
  • Journal Title: FEBS J. 276/5 Pages: 1418-1428
  • Peer Reviewed
• [Journal Article] Studies on epigenetic control of B cell functions using the DT40 cell line. (2009)
  • Author(s): Kikuchi, H., Barman, H. K., Nakayama, M., Takami, Y., Nakayama, T.
  • Journal Title: Immunogenetics Research Progress, Nova Science Publishers, Inc. NY (in press)
  • Peer Reviewed
• [Journal Article] クロマチン構築を介したDNA複製制御 (2009)
  • Author(s): 高見恭成
  • Journal Title: 蛋白質・核酸・酵素 54 Pages: 496-501
• [Journal Article] GCN5 and BCR signalling collaborate to induce pre-mature B cell apoptosis through depletion of ICAD and IAP2 and activation of caspase activities. (2008)
  • Author(s): Kikuchi, H., Nakayama, T.
  • Journal Title: Gene 419 Pages: 48-55
  • Peer Reviewed
• [Journal Article] Histone acetyltransferase-1 regulates integrity of cytosolic histone H3-H4 containing complex. (2008)
  • Author(s): Barman, H.K., Takami, Y., Nishijima, H., Shibahara, K., Sanematsu, F., Nakayama, T.
  • Journal Title: Biochem. Biophys. Res. Commun. 373 Pages: 624-630
  • Peer Reviewed
• [Journal Article] Histone acetyltransferase-1 regulates integrity of cytosolic histone H3-H4 containing complex. (2008)
  • Author(s): Barman, H. K., Takami, Y., Nishijima, H., Shibahara, K., Sanematsu, F., Nakayama, T.
  • Journal Title: Biochem. Biophys. Res. Commun. 373 Pages: 624-630
  • Peer Reviewed
• [Journal Article] Studies on epigenetic control of B cell functions using the DT40 cell line (2008)
  • Author(s): Kikuchi, H., Barman, H. K., Nakayama, M., Takami, Y. and Nakayama, T.
  • Journal Title: Immunogenetics Research Progress, Nova Science Publishers, Inc. NY (in press)
  • Peer Reviewed
• [Journal Article] Essential role of CAF-1-mediated rapid nucleosome assembly for DNA replication and cell division in vertebrate cells. (2007)
  • Author(s): Takami, Y., Ono, T., Fukagawa, T., Shibahara, K., Nakayama, T.
  • Journal Title: Mol. Biol. Cell 18 Pages: 129-141
  • Peer Reviewed
• [Journal Article] Collaborative roles of rH2AX and the Rad51 paralog Xrcc3 in homologous recombinational repair. (2007)
  • Author(s): Sonoda, E., Zhao, G.-Y., Kohzaki, M., Dhar, P.K., Kikuchi, K., Redon, C., Pilch, D.R., Bonner, W.W., Nakano, A., Watanabe, M., Nakayama, T., Takeda, S., Takami, Y.
  • Journal Title: DNA Repair 6 Pages: 280-292
  • Peer Reviewed
• [Journal Article] Cleavage and cytoplasmic relocalization of histone deacetylase 3 are important for apoptosis progression. (2007)
  • Author(s): Escaffit, F., Vaute, O., Chevillard-Briet, M., Segui, B., Takami, Y., Nakayama, T., Trouche, D.
  • Journal Title: Mol. Cell. Biol. 27 Pages: 554-567
  • Peer Reviewed
• [Journal Article] HDAC2 controls IgM H and L-chain gene expressions via EBF1, Pax5, Ikaros, Aiolos and E2A gene expressions. (2007)
  • Author(s): Nakayama, M., Suzuki, H., Yamamoto-Nagamatsu, N., Barman, H.K., Kikuchi, H., Takami, Y., Toyonaga, K., Yamashita, K., Nakayama, T.
  • Journal Title: Genes Cells 12 Pages: 359-373
  • Peer Reviewed
• [Journal Article] Histone H1 variant, H1R is involved in DNA damage response. (2007)
  • Author(s): Hashimoto, H., Sonoda, E., Takami, Y., Kimura, H., Nakayama, T., Tachibana, M., Takeda, S., Shinkai, Y.
  • Journal Title: DNA Repair 6 Pages: 1584-1595
  • Peer Reviewed
• [Journal Article] Essential role of CAF-1-mediated rapid nucleosome assembly for DNA replication and cell division in vertebrate cells (2007)
  • Author(s): Takami, Y., Ono, T., Fukagawa, T., Shibahara, K. and Nakayama, T.
  • Journal Title: Mol. Biol. Cell 18 Pages: 129-141
  • Peer Reviewed
• [Journal Article] Collaborative roles of rH2AX and the Rad51 paralog Xrcc3 in homologous recombinational repair (2007)
  • Author(s): Sonoda, E., Zhao, G.-Y., Kohzaki, M., Dhar, P. K., Kikuchi, K., Redon, C., Pilch, D. R., Bonner, W. W., Nakano, A., Watanabe, M., Nakayama, T., Takeda, S. and Takami, Y.
  • Journal Title: DNA Repair 6 Pages: 280-292
  • Peer Reviewed
• [Journal Article] Cleavage and cytoplasmic relocalization of histone deacetylase 3 are important for apoptosis progression (2007)
  • Author(s): Escaffit, F., Vaute, O., Chevillard-Briet, M., Segui, B., Takami, Y., Nakayama, T. and Trouche, D.
  • Journal Title: Mol. Cell. Biol. 27 Pages: 554-567
  • Peer Reviewed
• [Journal Article] HDAC2 controls IgM H and L-chain gene expressions via EBF1, Pax5, Ikaros, Aiolos and E2A gene expressions (2007)
  • Author(s): Nakayama, M., Suzuki, H., Yamamoto-Nagamatsu, N., Barman, H. K., Kikuchi, H., Takami, Y., Toyonaga, K., Yamashita, K. and Nakayama, T.
  • Journal Title: Genes Cells 12 Pages: 359-373
  • Peer Reviewed
• [Journal Article] Histone H1 variant, H1R is involved in DNA damage response (2007)
  • Author(s): Hashimoto, H., Sonoda, E., Takami, Y., Kimura, H., Nakayama, T., Tachibana, M., Takeda, S. and Shinkai, Y.
  • Journal Title: DNA Repair 6 Pages: 1584-1595
  • Peer Reviewed
• [Presentation] クルクミンによる白血球スーパーオキシド産生系誘導増強効果 (2010)
  • Author(s): 菊池秀彦、中山建男
  • Organizer: 第33回日本分子生物学会年会/第83回日本生化学会大会、合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2010-12-09
• [Presentation] ニワトリDT40細胞を用いたミトコンドリアSirtuinsの機能解析 (2010)
  • Author(s): 鈴木寛之、中山建男、高見恭成
  • Organizer: 第10回日本蛋白質科学会年会
  • Place of Presentation: 札幌
  • Year and Date: 2010-06-17
• [Presentation] Histone chaperone FACT is involved in efficient elongation of nascent DNA chain. (2009)
  • Author(s): 阿部拓也、大沢洸司、高見恭成、多田周右、中山建男、室伏廣、堀越正美、関政幸、榎本武美
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2009-12-12
• [Presentation] 転写因子AiolosによるBリンパ球アポトーシス制御機構の解析 (2009)
  • Author(s): 菊池秀彦、山下幸貴、中山雅美、豊永健二、中山建男
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2009-12-12
• [Presentation] ノックイン変異細胞を用いたASF1/CIAの機能解析 (2009)
  • Author(s): 高見恭成、中山建男
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2009-12-11
• [Presentation] GCN5による白血球スーパーオキシド産生系制御機構の解析 (2009)
  • Author(s): 木脇直美、菊池秀彦、栗林太、中山建男
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2009-12-09
• [Presentation] 転写因子E2AによるBリンパ球アポトーシス制御 (2009)
  • Author(s): 菊池秀彦、豊永健二、山下幸貴、中山雅美、中山建男
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸
  • Year and Date: 2009-10-23
• [Presentation] ノックイン変異ASF1/CIA細胞を用いたASF1の機能解析 (2009)
  • Author(s): 高見恭成、中山建男
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸
  • Year and Date: 2009-10-23
• [Presentation] 転写因子E2AによるBリンパ球アポトーシス制御機構の解析 (2009)
  • Author(s): 菊池秀彦、豊永健二、山下幸貴、中山雅美、中山建男
  • Organizer: 日本分子生物学会第9回シンポジウム
  • Place of Presentation: 宮崎
  • Year and Date: 2009-05-11
• [Presentation] ニワトリDT40細胞を用いたHat1の機能解析 (2009)
  • Author(s): 高見恭成、Hirak Kumar Barman, 中山建男
  • Organizer: 日本分子生物学会第9回シンポジウム
  • Place of Presentation: 宮崎
  • Year and Date: 2009-05-11
• [Presentation] ヒストンアセチルトランスフェラーゼGCN5によるBCRシグナリング制御 (2008)
  • Author(s): 菊池秀彦、中山建男
  • Organizer: 第31回日本分子生物学会年会、第81回日本生化学会大会、合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-10
• [Presentation] ニワトリDT40細胞を用いたhistone acethyltransferase-1(Hat1)の機能解析 (2008)
  • Author(s): 高見恭成、中山建男
  • Organizer: 第31回日本分子生物学会年会、第81回日本生化学会大会、合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-10
• [Presentation] ヒストンアセチルトランスフェラーゼGCN5によるBCRシグナリング制御. (2008)
  • Author(s): 菊池秀彦, 中山建男
  • Organizer: 第31回日本分子生物学会年会、第81回口本生化学会大会、合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-10
• [Presentation] ニワトリDT40細胞を用いたhistone acethyltransferase-1 (Hat1)の機能解析. (2008)
  • Author(s): 高見恭成, 中山建男
  • Organizer: 第31回日本分子生物学会年会、第81回日本生化学会大会、合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-10
• [Presentation] ニワトリDT40細胞を用いたhistone acethyltransferase-1(Hatl)の機能解析 (2008)
  • Author(s): Hirak Kumar Barman, 高見恭成、中山建男
  • Organizer: 平成20年度日本生化学会九州支部例会
  • Place of Presentation: 福岡
• [Presentation] ニワトリDT40細胞を用いたhistone acethyltransferase-1 (Hatl)の機能解析. (2008)
  • Author(s): Hirak Kumar Barman, 高見恭成, 中山建男
  • Organizer: 平成20年度日本生化学会九州支部例会
  • Place of Presentation: 福岡
• [Presentation] DT40細胞を用いた遺伝子ノックアウト法によるGCN5の機能解析 (2007)
  • Author(s): 菊池秀彦、高見恭成、中山建男
  • Organizer: 第30回日本分子生物学会年会、第80回日本生化学会大会、合同大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-13
• [Presentation] DT40細胞を用いたASF1の機能解析 (2007)
  • Author(s): 高見恭成、中山建男
  • Organizer: 第30回日本分子生物学会年会、第80回日本生化学会大会、合同大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-12
• [Presentation] DT40細胞を用いた遺伝子ノックアウト法によるGCN5の機能解析 (2007)
  • Author(s): 菊池秀彦、高見恭成、中山建男
  • Organizer: 日本生化学会九州支部例会
  • Place of Presentation: 宮崎
  • Year and Date: 2007-05-20
• [Presentation] 脱脂粉乳を利用した新規味噌の機能性 (2007)
  • Author(s): 六車三治男、伊藤瑞奈、河原聡、丸山眞杉、中山建男、久寿米木一裕、菱沼毅、中村豊郎
  • Organizer: 日本生化学会九州支部例会
  • Place of Presentation: 宮崎
  • Year and Date: 2007-05-20
• [Book] Studies on epigenetic control of B cell functions using the DT40 cell line.(Advances in Genetics Research Vol.2) (2010)
  • Author(s): Kikuchi, H., Barman, H.K., Nakayama, M., Takami, Y., Nakayama, T.
  • Publisher: Nova Science Publishers, Inc. NY
• [Book] Advances in Genetics Research vol.2 (2010)
  • Author(s): Kikuchi H
  • Publisher: Nova Science Publishers
• [Remarks] ホームページ等
  • URL: http://www.med.miyazaki-u.ac.jp/2bio/index.html
• [Patent(Industrial Property Rights)] レチノイン酸効果増強剤およびこれを用いたリンパ腫治療剤キット、並びに、活性酸素産生促進剤およびこれを用いた免疫賦活剤 (2010)
  • Inventor(s): 菊池秀彦、木脇直美、中山建男
  • Industrial Property Rights Holder: 宮崎大学
  • Industrial Property Number: 2010-059734
  • Filing Date: 2010-03-16