Regulation of DNA replication and repair by ubiquitin or ubiquitin-like modification in cancer cells

• Project/Area Number: 19590272
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Hamamatsu University School of Medicine
• Principal Investigator: UCHIDA Chiharu Hamamatsu University School of Medicine, 医学部, 助教 (60223567)
• Co-Investigator(Kenkyū-buntansha): 北川 雅敏 浜松医科大学, 医学部, 教授 (50294971)
• Co-Investigator(Renkei-kenkyūsha): KITAGAWA Masatoshi 浜松医科大学, 医学部, 教授 (50294971)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2008: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2007: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: pRB / クロマチン / ユビキチン / DNA複製 / がん化

Research Abstract

癌抑制遺伝子産物RBタンパク質(pRB)は、細胞増殖を正に制御しているE2F依存性の遺伝子発現を抑制する。最近pRBは細胞周期進行の抑制に働くのみならず、DNA複製制御因子との結合を介する複製反応阻害や、DNAダメージで誘導される細胞周期停止にも関与することが分かってきた。またDNAおよび染色体ヒストンのメチル化を介し、遺伝子発現を負に制御すると考えられているjumonjiは、pRBと結合することが知られている。これらはpRBと染色体(クロマチン)関連因子との相互作用が、適切な細胞機能の維持に重要であることを意味している。我々はpRBおよびpRB結合蛋白質のユビキチン様修飾とクロマチン制御との関連に注目し、次の2項目を目的として研究を進めた。1) RBタンパク質のユビキチン(様)修飾がDNA複製・修復関連因子との相互作用に関与し、複製・修復の過程を制御もしくは撹乱する可能性を検証する。2) pRBと結合する染色体制御関連因子のユビキチン化による細胞周期依存的な量的制御機構を解明し、がん化との関連を検証する。
本研究期間内では以下のような成果を得た。
(1) 2005年に我々はpRBがMdm2によってユビキチン化され、プロテアソーム依存的分解を受けることを報告した(Uchida et.al. EMBO J.)。本研究ではpRBが新規の修飾を受けることにより、pRB-E2F結合が抑制されることを見出した。この修飾はユビキチン様修飾因子の一つによるものであった。
(2) Mdm2はユビキチン化以外の働きによって、pRBの染色体への結合を制御している可能性が示された。
(3) jumonjiはMdm2によってユビキチン化されることが分かった。しかし、そのユビキチン化とjumonjiの安定性制御との関連性を示す結果は得られず、Mdm2によるjumonjiのユビキチン化は蛋白質分解以外のシグナルとなる可能性がある。

Research Products

• [Journal Article] TSU68 prevents liver metastasis of colon cancer xenografts by modulating the pre-metastatic niche (2008)
  • Author(s): Yamamoto, M., Kikuchi, H., Ohta, M., Kawabata, T., Hiramatsu, Y., Kondo, K., Baba, M., Kamiya, K., Tanaka, T., Kitagawa, M., Konno H.
  • Journal Title: Cancer Res 68 Pages: 9754-9762
  • Peer Reviewed
• [Journal Article] Intrarenal RAS activity and urinary angiotensinogen excretion in anti-thymocyte serum nephritis rats (2008)
  • Author(s): Ohashi, N., Yamamoto, T., Huang, Y., Misaki, T., Fukasawa, H., Suzuki, H., Togawa, A., Suzuki, S., Fujigaki, Y., Nakagawa, T., Nakamura, Y., Suzuki, F., Kitagawa, M., Hishida, A.
  • Journal Title: Am J.Physiol Renal Physiol 295 Pages: F1512-1518
  • Peer Reviewed
• [Journal Article] Novel insights into FGD3, a putative GEF for Cdc42, that undergoes SCFFWD1/ss-TrCP-mediated proteasomal degradation analogous to that of its homologue FGD1 but regulates cell morphology and motility differently from FGD1 (2008)
  • Author(s): Hayakawa, M., Matsushima, M.,Hagiwara, H., Oshima, T., Fujino, T., Ando, K., Kikugawa, K., Tanaka, H., Miyazawa,K., Kitagawa, M.
  • Journal Title: Genes to Cells 13 Pages: 329-442
  • Peer Reviewed
• [Journal Article] Pirh2 interacts with and ubiquitylates signal recognition particle receptor beta subunit (2008)
  • Author(s): Abe, K., Hattori, T., Isobe, T., Kitagawa, K., Oda, T., Uchida, C., Kitagawa, M.
  • Journal Title: Biomed Res 29 Pages: 53-60
  • Peer Reviewed
• [Journal Article] Stress via p53 pathway causes apoptosis by mitochondrial Noxa upregulation in doxorubicin-treated neuroblastoma cells (2008)
  • Author(s): Kurata, K., Yanagisawa, R., Ohira, M., Kitagawa, M., Nakagawara, A., Kamijo, T.
  • Journal Title: Oncogene 27 Pages: 741-754
  • Peer Reviewed
• [Journal Article] Regulation of TGF-beta signaling by Smads and its roles in tissue fibrosis (2008)
  • Author(s): Fukasawa, H., Yamamato, T., Kitagawa, M. and Hishida, A.
  • Journal Title: Current signal transduction therapy(review article) 3 Pages: 1-6
  • Peer Reviewed
• [Journal Article] TSU68 prevents liver metastasis of colon cancer xenografts by modulating the pre-metastatic niche. (2008)
  • Author(s): Yamamoto, M., et al.
  • Journal Title: Cancer Res. 68 Pages: 9754-9762
  • Peer Reviewed
• [Journal Article] Intrarenal RAS activity and urinary angiotensinogen excretion in anti-thymocyte serum nephritis rats. (2008)
  • Author(s): Ohashi, N., et al.
  • Journal Title: Am J. Physiol Renal Physiol. 295 Pages: F1512-1518
  • Peer Reviewed
• [Journal Article] Novel insights into FGD3, a putative GEF for Cdc42, that undergoes SCFFWDl/β-TrCP-mediated proteasomal degradation analogous to that of its homologue FGD1 but regulates cell morphology and motility differently from FGD1. (2008)
  • Author(s): Hayakawa, M., et al.
  • Journal Title: Genes to Cells 13 Pages: 329-442
  • Peer Reviewed
• [Journal Article] Pirh2 interacts with and ubiquitylates signal recognition particle recept or beta subunit (2008)
  • Author(s): Abe, K., et. al.
  • Journal Title: Biomed Res. 29 Pages: 53-60
  • Peer Reviewed
• [Journal Article] Pirh2 Promotes Ubiquitin-Dependent Degradation of the CDK Inhibitor p27Kip1 (2007)
  • Author(s): Hattori, T, Isobe, T, Abe, K, Kikuchi, H, Kitagawa, K, Oda, T, Uchida, C, Kitagawa, M.
  • Journal Title: Cancer Research 67 Pages: 10789-10795
  • Peer Reviewed
• [Journal Article] Critical and functional regulation of CHOP(C/EBP homologous protein)through the N-terminal portion (2007)
  • Author(s): Ohoka, N, Hattori, T, Kitagawa, M, Onozaki, K, and Hayashi H.
  • Journal Title: J Biol Chem 282 Pages: 35687-35694
  • Peer Reviewed
• [Journal Article] Contribution of Extracellular Signal-Regulated Kinases to the IL-1-induced Growth Inhibition of Human Melanoma Cells A375 (2007)
  • Author(s): Arakawa, T, Yamamura, T, Hattori, T, Hayashi H, Mori, A, Yoshida, A, Uchida, C, Kitagawa, M, Onozaki, K.
  • Journal Title: International Immunopharmacology 8 Pages: 80-89
• [Journal Article] Cys-881 is essential for the trafficking and secretion of truncated mutant ceruloplasmin in aceruloplasminemia (2007)
  • Author(s): Kono, S, Suzuki, H, Oda, T, Takahashi, Y, Shirakawa, K, Takahashi, Y, Kitagawa, M, Miyajima, H.
  • Journal Title: J.Hepatology 47 Pages: 844-850
  • Peer Reviewed
• [Journal Article] Effect of loss of heterozygosity of the c-kit gene on prognosis after hepatectomy for metastatic liver gastrointestinal stromal tumors (2007)
  • Author(s): Kikuchi, H, Yamamoto, M, Hiramatsu, Y, Baba, M, Ohta, M, Kamiya, K, Tanaka, T, Suzuki, S, Sugimura, H, Kitagawa, M, Kanai, T, Kitayama, Y, Kanda, T, Nishikura, K, Konno, H.
  • Journal Title: Cancer Sci 98 Pages: 1734-1739
  • Peer Reviewed
• [Journal Article] ARA54 is involved in transcriptional regulation of the cyclin D1 gene in human cancer cells (2007)
  • Author(s): Kikuchi, H, Uchida, C, Hattori, T, Isobe, T, Hiramatsu, Y, Kitagawa, K, Oda, T, Konno, H, Kitagawa, M.
  • Journal Title: Carcinogenesis 28 Pages: 1752-1758
  • Peer Reviewed
• [Journal Article] Renal damage in obstructive nephropathy is decreased in Skp2-deficient mice. Am (2007)
  • Author(s): Suzuki, S, Fukasawa, H, Kitagawa, K, Uchida, C, Hattori, T, Isobe, T, Oda, T, Misaki, T, Ohashi, N, Nakayama, K, Nakayama, K, I., Hishida, A, Yamamoto, T, Kitagawa, M.
  • Journal Title: J.Pathol 171 Pages: 473-483
  • Peer Reviewed
• [Journal Article] Phosphorylation of pRB at Ser612 by Chk1/2 leads to a complex between pRB and E2F-1 after DNA damage (2007)
  • Author(s): Inoue, Y, Kitagawa, M, Taya, Y.
  • Journal Title: EMBO J 26 Pages: 2083-2093
  • Peer Reviewed
• [Journal Article] Genetically engineered neural stem cells migrate and suppress glioma cell growth at distant intracranial sites (2007)
  • Author(s): Li, S, Gao, Y, Tokuyama, T, Yamamoto, J, Yokota, N, Yamamoto, S, Terakawa, S, Kitagawa, M, Namba, H.
  • Journal Title: Cancer Letters 251(2) Pages: 220-227
  • Peer Reviewed
• [Journal Article] Pirh2 Promotes Ubiquitin-Dependent Degradation of the CDK Inhibitor p27Kip1 (2007)
  • Author(s): Hattori, T., et. al.
  • Journal Title: Cancer Res. 67 Pages: 10789-10795
  • Peer Reviewed
• [Journal Article] ARA54 is involved in transcriptional regulation of the cyclin D1 gene in human cancer cells (2007)
  • Author(s): Kikuchi, H., et. al.
  • Journal Title: Carcinogenesis 28 Pages: 1752-1758
  • Peer Reviewed
• [Journal Article] Renal damage in obstructive nephropathy is decreased in Skp2-deficient mice (2007)
  • Author(s): Suzuki, S., et. al.
  • Journal Title: Am.J.Pathol 171 Pages: 473-483
  • Peer Reviewed
• [Journal Article] Effect of loss of heterozygosity of the c-kit gene on prognosis after hepateCtOmy for metastatic liver gastrointestinal stromal tumors (2007)
  • Author(s): Kikuchi, H., et. al.
  • Journal Title: Cancer Sci. 98 Pages: 1734-1739
  • Peer Reviewed
• [Journal Article] Cys-881 is essential for the trafficking and secretion of truncated mutant ceruloplasmin in aceruloplasminemia (2007)
  • Author(s): Kono, S., et. al.
  • Journal Title: J.Hepatology 47 Pages: 844-850
  • Peer Reviewed
• [Journal Article] Stress via p53 pathway causes apoptosis by mitochondrial Noxa upregulation in doxorubicin-treated neuroblastoma cells (2007)
  • Author(s): Kurata, K., et. al.
  • Journal Title: Oncogene 27 Pages: 741-754
  • Peer Reviewed
• [Journal Article] Critical and functional regulation of CHOP (C/EBP homologous protein)through the N-terminal al portion (2007)
  • Author(s): Ohoka, N., et. al.
  • Journal Title: J Biol Chem. 282 Pages: 35687-35694
  • Peer Reviewed
• [Journal Article] Contribution of Extracellular Signal-Regulated Kinases to theI IL-1-induced Growth Inhibition of Human Melanoma Cells A375 (2007)
  • Author(s): Arakawa, T., et. al.
  • Journal Title: International lmmunopharmacology 8 Pages: 80-89
  • Peer Reviewed
• [Presentation] GPR48による癌転移機構の解析 (2008)
  • Author(s): 磯部智康, 島田真衣, 劉寧, 北川恭子, 鈴木小由里, 小田敏明, 服部隆行, 内田千晴, 北川雅敏
  • Organizer: 第31回日本分子生物学会、第81回日本生化学会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-10
• [Presentation] タンパク質過剰発現誘導性小胞体の構造と機能 (2008)
  • Author(s): 小田敏明, 服部隆行, 磯部智康, 北川恭子, 内田千晴, 横田貞記, 北川雅敏
  • Organizer: 第31回日本分子生物学会、第81回日本生化学会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-09
• [Presentation] 細胞周期制御分子p27のユビキチン依存的分解機構と病態 (2007)
  • Author(s): 北川雅敏
  • Organizer: 第30回日本分子生物学会、第80回日本生化学会合同大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-12
• [Presentation] 細胞周期制御分子p27のユビキチン依存的分解機構と病態 (2007)
  • Author(s): 北川 雅敏
  • Organizer: 第30回日本分子生物学会第80回日本生化学会合同大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-12
• [Presentation] 腎障害進行におけるSkp2発現充進の解析;Skp2K:0マウスを用いた検討 (2007)
  • Author(s): 鈴木 小由里
  • Organizer: 第30回日本分子生物学会第80回日本生化学会合同大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-12
• [Presentation] 腎障害進行におけるSkp2発現亢進の解析 : Skp2KOマウスを用いた検討 (2007)
  • Author(s): 鈴木小由里, 深澤洋敬, 北川恭子, 戸川証大橋温, 小田敏明, 内田千晴, 服部隆行, 中山敬一, 山本龍夫, 菱田明, 北川雅敏
  • Organizer: 第30回日本分子生物学会、第80回日本生化学会合同大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-07
• [Presentation] Novel regulatory mechanisms of Cyclin D1 expression (2007)
  • Author(s): Kitagawa, M, et.al.
  • Organizer: 第66回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2007-10-05
• [Presentation] 癌関連核内因子のユビキチン依存的制御機構 (2007)
  • Author(s): 北川雅敏
  • Organizer: Nulear Signaling JAPAN 2007シンポジウム
  • Place of Presentation: 東京
  • Year and Date: 2007-07-13
• [Presentation] 癌関連核内因子のユビキチン依存的制御機構 (2007)
  • Author(s): 北川 雅敏
  • Organizer: Nuclear Signaling JAPAN2007シンポジウム
  • Place of Presentation: 東京
  • Year and Date: 2007-07-13
• [Remarks]
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• [Remarks]
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• [Remarks]
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