The analysis of basic chromosomal functions using human artificial chromosomes (HACs) and the development of novel HACs.

• Project/Area Number: 20247019
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Molecular biology
• Research Institution: Kazusa DNA Research Institute
• Principal Investigator: MASUMOTO Hiroshi Kazusa DNA Research Institute, ヒトゲノム研究部, 室長 (70229384)
• Co-Investigator(Renkei-kenkyūsha): NAKANO Megumi (財団法人)かずさDNA研究所, 研究員 (50542825) ; OHZEKI Jun-Ichirou (財団法人)かずさDNA研究所, 研究員 (30514088)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥44,070,000 (Direct Cost: ¥33,900,000、Indirect Cost: ¥10,170,000); Fiscal Year 2010: ¥9,750,000 (Direct Cost: ¥7,500,000、Indirect Cost: ¥2,250,000); Fiscal Year 2009: ¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000); Fiscal Year 2008: ¥22,620,000 (Direct Cost: ¥17,400,000、Indirect Cost: ¥5,220,000)
• Keywords: 遺伝学 / 染色体 / 分配 / セントロメア / 人工染色体 / クロマチン / エピジェネティックス / ヘテロクロマチン

Research Abstract

We have developed a novel human artificial chromosome (HAC) system to investigate the epigenetic state of chromatin by applying a fusion-protein tethering system. We found that altering the HAC centromere chromatin to a more heterochromatic with the tTS transcription silencer or histone methyl transferase (Suv39h) caused mis-segregation of the HAC by losing kinetochore structure. Otherwise, histone acetyl transferase (HAT) fusions enhanced hyper-assembly of kinetocore components, indicating that a dynamic counterbalance is essential for mammalian kinetochore assembly.

Research Products

• [Journal Article] Epigenetic engineering sho ws H3K4 me2 is required for HJURP targeting and CENP-A assembly on a synthetic human kinetochore. (2011)
  • Author(s): Bergmann J.H., Gomez M., Martins N.M.C., Kimura H., Kelly D.A., Masumo to H., Larionov V., Jansen L.E.T., Earnshaw W.C.
  • Journal Title: EMBO J. 30 Pages: 328-340
  • Peer Reviewed
• [Journal Article] Epigenetic engineering shows H3K4me2 is required for HJURP targeting and CENP-A assembly on a synthetic human kinetochore (2011)
  • Author(s): Jan H.Bergmann
  • Journal Title: EMBO J. Volume: 30 Pages: 328-340
  • Peer Reviewed
• [Journal Article] Human Artificial Chromosome with a Conditional Centromere for Gene Delivery and Gene Expression (2010)
  • Author(s): Iida Y., Kim JH., Kazuki Y., Hoshiya H., Takiguchi M., Hayashi M., Erliandri I., Lee HS., Samoshkin A., Masumoto H., Earnshaw WC., Kouprina N., Larionov V., Oshimura M.
  • Journal Title: DNA Res. 17 Pages: 293-301
  • Peer Reviewed
• [Journal Article] Human Artificial Chromosome with a Conditional Centromere for Gene Delivery and Gene Expression (2010)
  • Author(s): Yuichi Iida
  • Journal Title: DNA RESEARCH Volume: 17 Pages: 293-301
  • Peer Reviewed
• [Journal Article] Hierarchical inactivation of a synthetic human kinetochore by a chromatin modifier. (2009)
  • Author(s): Cardinale S., Bergmann J.H., Kelly D., Nakano M., Valdivia M.M., Kimura H., Masumoto H., Larionov V., Earnshaw W.C.
  • Journal Title: Mol.Biol.Cell 20 Pages: 4194-4204
  • Peer Reviewed
• [Journal Article] Human gamma-satellite DNA maintains open chromatin structure and protects a transgene from epigenetic silencing. (2009)
  • Author(s): Kim JH., Ebersole T., Kouprina N., Noskov V.N., Ohzeki J., Masumoto H., Mravinac B., Sullivan B.A., Pavlicek A., Dovat S., Pack S.D., Kwon YW., Flanagan P.T., Loukinov D., Lobanenkov V., Larionov V.
  • Journal Title: Genome Res. 19 Pages: 533-544
  • Peer Reviewed
• [Journal Article] Hierarchical inactivation of aynthetic human kinetochore by a chromatin modifier (2009)
  • Author(s): Stefano Cardinale
  • Journal Title: Molecular Biology of the Cell 20 Pages: 4194-4204
  • Peer Reviewed
• [Journal Article] Human gamma-satellite DNA maintains open chromatin structure and protects a transgene from epigenetic silencing (2009)
  • Author(s): Jung-Hyun Kim
  • Journal Title: Genome Research 19 Pages: 533-544
  • Peer Reviewed
• [Journal Article] 染色体分配装置セントロメアの形成メカニズム (2009)
  • Author(s): 中野めぐみ
  • Journal Title: 蛋白質核酸酵素 54 (4) Pages: 427-435
• [Journal Article] Human gamma-satellite DNA maintains open chromatin structure and protects a transgene from epigenetic silencing (2009)
  • Author(s): Jung-Hyun Kim
  • Journal Title: Geome Research 19 Pages: 533-544
  • Peer Reviewed
• [Journal Article] 染色体分配装置セントロメアの形成メカニズム (2009)
  • Author(s): 中野めぐみ
  • Journal Title: 蛋白質核酸酵素 54(4) Pages: 427-435
• [Journal Article] Inactivation of a human kinetochore by specific targeting of chromatin modifiers. (2008)
  • Author(s): Nakano M., Cardinale S., Noskov V.N., Gassmann R., Vagnarelli P., Kandels-Lewis S., Larionov V., Earnshaw W.C., Masumoto H.
  • Journal Title: Dev.Cell 14 Pages: 507-522
  • Peer Reviewed
• [Journal Article] Inactivation of a human kinetochore by specific targeting of chromatin modifiers (2008)
  • Author(s): Megumi Nakano
  • Journal Title: Developmental Cell 14 Pages: 507-522
  • Peer Reviewed
• [Journal Article] ヒト人工染色体から見た染色体制御機構 (2008)
  • Author(s): 舛本寛
  • Journal Title: 生化学 80 Pages: 200-209
  • NAID: 10021191161
• [Presentation] A chromatin assembly balance determines de novo kinetochre formation on satellite DNA. (2010)
  • Author(s): Hiroshi Masumoto
  • Organizer: The MEXT International Workshop 'Mitosis : Cell Proliferation Control
  • Place of Presentation: Tokyo
  • Year and Date: 2010-11-08
• [Presentation] コンディショナルセントロメアを持つ人工染色体 (2010)
  • Author(s): 舛本寛
  • Organizer: 第82回日本遺伝学会大会
  • Place of Presentation: 札幌
  • Year and Date: 2010-09-22
• [Presentation] A chromatin assembly balance determines the fate of de novo kinetochre or heterochromatin formation on satellite DNA. (2010)
  • Author(s): Masumoto H., Ohzeki J., Nakano M., et al. V.
  • Organizer: EMBO Workshop on Chromosome Segregation and Aneuploidy
  • Place of Presentation: Edinburgh
• [Presentation] A Dynamic Assembly Balance of Centromere Chromatin and Heterochromatin on Alpha-satellite DNA. (2009)
  • Author(s): 舛本寛
  • Organizer: The American Society of Human Genetics The 59th Annual Meeting, in The Invited Session "Epigenetics of human centromere formation"
  • Place of Presentation: Honolulu, Hawaii
  • Year and Date: 2009-10-20
• [Presentation] A Dynamic Assembly Balance of Centromere Chromatin and Heterochro matin on Alpha-satellite DNA (2009)
  • Author(s): Hiroshi Masumoto
  • Organizer: The American Society of Human Genetics The 59th Annual Meeting
  • Place of Presentation: Honolulu, USA
  • Year and Date: 2009-10-20
• [Presentation] Dynamic Assembly mechanism of Centromere Chromatin and Heterochromatin on Human Satellite DNA. (2009)
  • Author(s): Masumoto H., Ohzeki J., Nakano M., Okada T., Larionov V., Earnshaw W.C.
  • Organizer: The 24^<th> Naito Conference on "Nuclear Dynamics and RNA [II]"
  • Place of Presentation: Sapporo
  • Year and Date: 2009-06-25
• [Presentation] Dynamic Assembly mechanism of Centromere Chromatin and Heterochromatin on Human Satellite DNA (2009)
  • Author(s): Hiroshi Masumoto
  • Organizer: The 24^<th> Naito Conference on "Nuclear Dynamics and RNA [II]"
  • Place of Presentation: Sapporo, Japan
  • Year and Date: 2009-06-25
• [Presentation] A Dynamic Assembly Balance of CENP-A Chromatin or Heterochromatin for Centromere Activity on Alpha-satellite Repeats. (2008)
  • Author(s): 舛本寛, et al.
  • Organizer: EMBO Workshop Chromosome Segregation : Centromeres and Kinetochores
  • Place of Presentation: Arcachon, Bordeaux, France
  • Year and Date: 2008-09-28
• [Presentation] A Dynamic Assembly Balance of CENP-A Chromatin or Heterochromatin for Centromere Activity on Alpha-satellite Repeats (2008)
  • Author(s): Hiroshi Masumoto
  • Organizer: EMBO Workshop : Chromosome Segregation : Centromeres and Kinetochores
  • Place of Presentation: Bordeaux, France
  • Year and Date: 2008-09-28
• [Presentation] Dynamic Assembly and Inactivation of Human Centromere on Satellite DNA (2008)
  • Author(s): 舛本寛, et al.
  • Organizer: International Symposium on Chromosome Dynamics in Ise
  • Year and Date: 2008-05-28
• [Presentation] DYNAMIC ASSEMBLY AND INACTIVATION OF HUMAN CENTROMERE ON SATELLITE DNA (2008)
  • Author(s): Hiroshi Masumoto
  • Organizer: International Symposium on Chromosome
  • Place of Presentation: Ise, Japan
  • Year and Date: 2008-05-28
• [Presentation] セントロメア機能形成に関わるダイナミックなクロマチン集合バランス (2008)
  • Author(s): 舛本寛、中野めぐみ、大関淳一郎、他
  • Organizer: 第31回日本分子生物学会年会
  • Place of Presentation: 神戸
• [Book] 染色体分配装置セントロメアの形成メカニズム、蛋白質核酸酵素(54 査読無し) (2009)
  • Author(s): 中野めぐみ、舛本寛
• [Book] The Kinetochore : from Molecular Discoveries to Cancer Therapy(Eds. De Wulf P., and Earnshaw W.C.)(Human Artificial Centromeres : De novo assembly of functional centromeres on human artificial chromosomes. 査読あり) (2008)
  • Author(s): Masumoto H., Okada T., Okamoto Y.
  • Publisher: Springer Publ.New York
• [Book] ヒト人工染色体から見た染色体制御機構(80 査読無し) (2008)
  • Author(s): 舛本寛、岡田晃明
  • Publisher: 生化学
• [Book] Human Artificial Centromeres : De novo assembly of functional centromeres on human artificial chromosomes. in "The Kinetochore : from Molecular Discoveries to Cancer Therapy". Eds. De Wulf P., and Earnshaw W. C. (2008)
  • Author(s): Hiroshi Masumoto
  • Total Pages: 26
  • Publisher: Springer-Verlag
• [Remarks] ホームページ等
  • URL: http://www.kazusa.or.jp/j/laboratories/labo_cellengineering.html