Molecular mechanisms for pre-replication complexes assembly and its implication in chromosomal stability in mammalian cells

2009 Fiscal Year Final Research Report

• Project/Area Number: 17080013
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: National Cancer Center Research Institute and Research Center for Innovative Oncology, National Cancer Center Hospital East
• Principal Investigator: FUJITA Masatoshi National Cancer Center Research Institute and Research Center for Innovative Oncology, National Cancer Center Hospital East, ウイルス部, 室長 (30270713)
• Co-Investigator(Kenkyū-buntansha): NISITANI Hidbo 兵庫県立大学, 大学院・生命理学, 教授 (40253455) ; MIZUNO Takeshi 理化学研究所, 今本細胞核機能, 専任研究員 (30281629)
• Project Period (FY): 2005 – 2009
• Keywords: 染色体DNA複製 / 細胞周期 / クロマチン / 染色体安定性

Research Abstract

Chromosomal DNA, an essential genetic element, should be precisely replicated during cell growth, failure of which leads to cell death or cancer development. In this research project, we have clarified molecular mechanisms by which DNA replicates once and only once during cells dividing into two daughter cells. Especially, we found that Cdt1 protein is strictly regulated by multiple mechanisms, which are essential for precise DNA replication. Base on these findings, we think that Cdt1 would be a novel target of anti-cancer drugs and we are now trying to find out such compounds. In addition, it turned out that such mechanisms for DNA replication also contribute to maintenance of other chromosomal functions.

Research Products

• [Journal Article] CDC6 interaction with ATR regulates activation of a replication checkpoint in higher eukaryotic cells. (2010)
  • Author(s): Yoshida K, Sugimoto N, Iwahori S, Yugawa T, Narisawa-Saito M, Kiyono T, Fujita M.
  • Journal Title: J. C e l l S c i. 123 Pages: 225-235
• [Journal Article] Redundant and differential regulation of multiple licensing factors ensures prevention of rereplication in normal human cells. (2009)
  • Author(s): Sugimoto N, Yoshida K, Tatsumi Y, Yugawa T, Narisawa-Saito M, Waga S, Kiyono T, Fujita M.
  • Journal Title: J. C e l l S c i. 122 Pages: 1184-1191
• [Journal Article] Cyclin A-Cdk1 regulates the origin firing program in mammalian cells. (2009)
  • Author(s): Katsuno Y, Suzuki A, Sugimura K, Okumura K, Zineldeen DH, Shimada M, Niida H, Mizuno T, Hanaoka F, Nakanishi M.
  • Journal Title: P r o c. N a t l. A c a d. S c i. U S A 106 Pages: 3184-3189
• [Journal Article] Identification of novel human Cdt1-binding proteins by a proteomics approach: Proteolytic regulation by APC/CCdh1. (2008)
  • Author(s): Sugimoto N, Kitabayashi I, Osano S, Tatsumi Y, Yugawa T, Narisawa-Saito M, Matsukage A, Kiyono T, Fujita, M.
  • Journal Title: M o l. B i o l. C e l l 19 Pages: 1007-1021
• [Journal Article] The inhibitory action of SQDG (sulfoquinovosyl diacylglycerol) from spinach on Cdt1-geminin interaction. (2008)
  • Author(s): Mizushina Y, Takeuchi T, Hada T, Maeda N, Sugawara F, Yoshida H, Fujita M.
  • Journal Title: B i o c h i m i e 90 Pages: 947-956
• [Journal Article] Coenzyme Q10 as a potent compound that inhibits Cdt1-geminin interaction. (2008)
  • Author(s): Mizushina Y, Takeuchi T, Takakusagi Y, Yonezawa Y, Mizuno T, Yanagi K, Imamoto N, Sugawara F, Sakaguchi K, Yoshida H, Fujita M.
  • Journal Title: B i o c h i m. B i o p h y s. A c t a. 1780 Pages: 203-213
• [Journal Article] Involvement of human ORC and TRF2 in pre-replication complex assembly at telomeres. (2008)
  • Author(s): Tatsumi Y, Ezura K, Yoshida K, Yugawa T, Narisawa-Saito M, Kiyono T, Ohta S, Obuse C, Fujita M.
  • Journal Title: G e n e s C e l l s 13 Pages: 1045-1059
• [Journal Article] CDK inhibitor p21 is degraded by a PCNA coupled Cul4-DDB1Cdt2 pathway during S phase and after UV irradiation. (2008)
  • Author(s): Nishitani H, Shiomi Y, Iida H, Michishita M, Takami T, Tsurimoto T.
  • Journal Title: J. B i o l. C h e m. 283 Pages: 29045-29052
• [Journal Article] Cdt1 associates dynamically with chromatin throughout G1 and recruits Geminin onto chromatin. (2007)
  • Author(s): Xouri G, Squire A, Dimaki M, Geverts B, Verveer PJ, Taraviras S, Nishitani H, Houtsmuller AB, Bastiaens PI, Lygerou Z.
  • Journal Title: E M B O J. 26 Pages: 1303-1314
• [Journal Article] Deregulation of Cdt1 induces chromosomal damage without rereplication and leads to chromosomal instability. (2006)
  • Author(s): Tatsumi Y, Sugimoto N, Yugawa T, Narisawa-Saito M, Kiyono T, Fujita M.
  • Journal Title: J. C e l l S c i. 119 Pages: 3128-3140
• [Journal Article] wo E3 ubiquitin ligases, SCF-Skp2 and DDB1-Cul4, target human Cdt1 for proteolysis. (2006)
  • Author(s): Nishitani H, Sugimoto N, Roukos V, Nakanishi Y, Saijo M, Obuse C, Tsurimoto T, Nakayama K-I, Nakayama K, Fujita M, Lygerous Z, Nishimoto T.
  • Journal Title: E M B O J. 25 Pages: 1126-1136
• [Journal Article] Cdt1 revisited: complex and tight regulation during the cell cycle and consequences of deregulation in mammalian cells. (2006)
  • Author(s): Fujita M.
  • Journal Title: C e l l D i v. 1 Pages: 22
• [Journal Article] Cdt1 phosphorylation by cyclin A-dependent kinases negatively regulates its function without affecting geminin binding. (2004)
  • Author(s): Sugimoto N, Tatsumi Y, Tsurumi T, Matsukage A, Kiyono T, Nishitani H, Fujita M.
  • Journal Title: J. B i o l. C h e m. 279 Pages: 19691-19697
• [Journal Article] Structure and mutagenesis studies of the C-terminal region of licensing factor Cdt1 enable the identification of key residues for binding to replicative helicase MCM proteins J.
  • Author(s): J. -G. Jee, Mizuno T, Kamada K, Tochio H, Chiba Y, Yanagi K, Yasuda G, Hiroaki H, Hanaoka F, Shirakawa M.
  • Journal Title: B i o l. C h e m.
• [Presentation] CDC6 interaction with ATR regulates activation of a replication checkpoint in higher eukaryotic cells. (2009)
  • Author(s): Fujita M.
  • Organizer: I n t e r n a t i o n a l S y m p o s i u m o n C h r o m o s o m e C y c l e a n d G n o m e D y n a m i c s.
  • Place of Presentation: Nasu, Japan.
  • Year and Date: 2009-11-11
• [Presentation] A chromatin remodeler SNF2H is involved in pre-RC formation and Cdt1-induced checkpoint activation. (2009)
  • Author(s): Sugimoto N, Waga S, Kiyono T, Fujita, M.
  • Organizer: K e y s t o n e S y m p o s i a " T e l o m e r e B i o l o g y a n d D N A R e p a i r"
  • Place of Presentation: Ashmore, Queensland, Australia.
  • Year and Date: 2009-10-11
• [Presentation] Cdk phosphorylation-dependent interaction of human CDC6 with ATR is involved in G2/M checkpoint response. (2008)
  • Author(s): Yoshida K, Kiyono T, Fujita M.
  • Organizer: T h e M e e t i n g o n D N A R e p l i c a t i o n a n d G e n o m e I n t e g r i t y a t S a l k I n s t i t u t e
  • Place of Presentation: La Jolla, USA.
  • Year and Date: 2008-07-18
• [Presentation] Involvement of human ORC and TRF2 in pre-replication complex assembly at telomeres and telomere homeostasis. (2007)
  • Author(s): Tatsumi Y, Ezura K, Yoshida K, Kiyono T, Ohta S, Obuse C, Fujita, M.
  • Organizer: C o l d S p r i n g H a r b o r L a b o r a t o r y M e e t i n g o n E u k a r y o t i c D N A R e p l i c a t i o n & G e n o m e M a i n t e n a n c e
  • Place of Presentation: Cold Spring Harbor, USA.
  • Year and Date: 2007-09-06
• [Book] 蛋白質核酸酵素(正井久雄、升方久夫、釣本敏樹、仁木宏典、篠原彰編集, 染色体動態制御における複製開始複合体因子の新たな機能, 増刊号「染色体サイクル」) (2009)
  • Author(s): 藤田雅俊.
  • Total Pages: 531-536
  • Publisher: 共立出版
• [Book] Encylopedic Reference of Genomics and Proteomics in Molecular Medicine(edited by Ganten, D. and Ruckpaul, K., DNA Replication Initiation.). (2006)
  • Author(s): Fujita M.
  • Total Pages: 446-449
  • Publisher: Springer
• [Remarks] 現所属、九州大学・大学院薬学研究院ホームページ
  • URL: http://tansaku.phar.kyushu-u.ac.jp/saito/top.html
• [Patent(Industrial Property Rights)] 抗癌性物質のスクリーニング方法 (2006)
  • Inventor(s): 藤田雅俊、水品善之
  • Industrial Property Rights Holder: 財団法人ヒューマンサイエンス振興財団
  • Industrial Property Number: 特願2006-281754
  • Filing Date: 2006-10-16