A novel proteomics approach for identification of protein-RNA complex associated with specific gene loci using a human artificial chromosome (HAC)

2011 Fiscal Year Final Research Report

• Project/Area Number: 21200038
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research a proposed research project)
• Allocation Type: Single-year Grants
• Research Field: Applied genomics; Genetics/Genome dynamics
• Research Institution: Tottori University
• Principal Investigator: KUGOH Hiroyuki 鳥取大学, 大学院・医学系研究科, 准教授 (40225131)
• Co-Investigator(Kenkyū-buntansha): NAKAYAMA Yuji 鳥取大学, 生命機能研究支援センター, 助教 (40432603)
• Project Period (FY): 2009 – 2011
• Keywords: 染色体工学

Research Abstract

To recover the CHO chromosome that carry 3.5kb of the lacO repeat sequences in the cells, we carried out immunoprecipitation by using anti-GFP antibody after the introduction of the LacI-GFP vector. As a result, a large size of CHO chromosome was recovered by anti-GFP antibody. These results suggest that the possibility of recovery of the HAC by chromatin immunoprecipitation. In addition, our colleague has developed a new HAC vector (MI-HAC) with multi-integrase recombination sites. MI-HAC enables to introduce various large genes or genomic loci. Furthermore, we produced MI-HAC that contains the lacO repeat sequences and targeted gene loci (Xist, LIT1/KCNQ1OT1 and CGG repeat sequence).

Research Products

• [Journal Article] Electrophysiological properties of prion-positive cardiac progenitors derived from murine embryonic stem cells (2012)
  • Author(s): Fujii H, Ikeuchi Y, Kurata Y, Ikeda N, Bahrudin U, Li P, Nakayama Y, Endo R, Hasegawa A, Morikawa K, Miake J, Yoshida A, Hidaka K, Morisaki T, Ninomiya H, Shirayoshi Y, Yamamoto K, Hisatome I
  • Journal Title: Circ J Volume: 76(12) Pages: 2875-83
  • Peer Reviewed
• [Journal Article] Down-regulation of MutS homolog 3 by hypoxia in human colorectal cancer (2012)
  • Author(s): Li J, Koike J, Kugoh H, Arita M, Ohhira T, Kikuchi Y, Funahashi K, Takamatsu K, Boland CR, Koi M, Hemmi H
  • Journal Title: Biochim Biophys Acta -Molecular Cell Research Volume: 1823(4) Pages: 889-899
  • DOI: doi:10.1016/ j.bbamcr.2012.01.017
  • Peer Reviewed
• [Journal Article] A method for producing transgenic cells using a multi-integrase system on a human artificial chromosome vector (2011)
  • Author(s): Yamaguchi S, Kazuki Y, Nakayama Y, Nanba E, Oshimura M, Ohbayashi T
  • Journal Title: PLoS One Volume: 6 Pages: e17267
  • Peer Reviewed
• [Journal Article] SIRT2 downregulation in HeLa can induce p53 accumulation via p38 MAPK activation-dependent p300 decrease, eventually leading to apoptosis (2011)
  • Author(s): Li YC, Matsumori H, Nakayama Y, Osaki M, Kojima H, Kurimasa A, Ito H, Mori S, Katoh M, Oshimura M, and Inoue T
  • Journal Title: Genes to Cells Volume: 16 Pages: 34-45
  • Peer Reviewed
• [Journal Article] Dysregulation of Gene Expression in the Artificial Human Trisomy Cells of Chromosome 8 Associated with Transformed Cell Phenotypes (2011)
  • Author(s): Nawata H, Genro Kashino G, Tano K, Daino K, Shimada Y, H Kugoh H, Oshimura M, Watanabe M
  • Journal Title: PLoS ONE Volume: 6(9) Pages: e25319
  • DOI: doi:10.1371/ journal.pone.0025319
  • Peer Reviewed
• [Journal Article] Identification of PITX1 as a TERT suppressor gene located on human chromosome 5 (2011)
  • Author(s): Qi DL, Ohhira T, Fujisaki C, Inoue T, Ohta T, Osaki M, Ohshiro E, Seko T, Aoki S, Oshimura M & Kugoh H
  • Journal Title: Mol. Cell Biol Volume: 31(8) Pages: 1624-1636
  • DOI: doi:10.1128/ MCB.00470-10
  • Peer Reviewed
• [Journal Article] Human chromosome 5 carries a transcriptional regulator of human telomerase reverse transcriptase (hTERT) (2010)
  • Author(s): Qi DL, Ohhira T, Oshimura M, Kugoh H
  • Journal Title: Biochem Biophys Res Commun Volume: 398(4) Pages: 695-701
  • DOI: doi:10.1016/ j.bbrc.2010.07.003
  • Peer Reviewed
• [Journal Article] Exploitation of the interaction of measles virus fusogenic envelope proteins with the surface receptor CD46 on human cells for microcell-mediated chromosome transfer (2010)
  • Author(s): Katoh M, Kazuki Y, Kazuki K, Kajitani N, Takiguchi M, Nakayama Y, Nakamura T, Oshimura M
  • Journal Title: BMC Biotechnology Volume: 10 Pages: 37
  • Peer Reviewed
• [Journal Article] Localization of an hTERT repressor region on human chromosome 3p21.3 using chromosome engineering (2010)
  • Author(s): Abe S., Tanaka H., Notsu T., Horike S., Fujisaki C., Qi DL., Ohhira T., Gilley D., Oshimura M., Kugoh H
  • Journal Title: Genome Integrity Volume: 1(1) Pages: 6
  • DOI: doi:10.1186/ 2041-9414-1-6
  • Peer Reviewed
• [Journal Article] SIRT2 downregulation confers resistance to microtubule inhibitors by prolonging chronic mitotic arrest (2009)
  • Author(s): Inoue T, Nakayama Y, Yamada H, Li YC, Yamaguchi S, Osaki M, Kurimasa A, Hiratsuka M, Katoh M, Oshimura M
  • Journal Title: Cell Cycle Volume: 8(8) Pages: 1279-91
  • Peer Reviewed
• [Journal Article] Identification, isolation and characterization of HCN4-positive pacemaking cells derived from murine embryonic stem cells during cardiac differentiation (2009)
  • Author(s): Morikawa K, Bahrudin U, Miake J, Igawa O, Kurata Y, Nakayama Y, Shirayoshi Y, Hisatome I
  • Journal Title: PACE Volume: PACE-09-0353 Pages: R1
  • Peer Reviewed
• [Journal Article] Identification of the chromatin regions coated by non-coding Xist RNA (2009)
  • Author(s): Murakami, K., Ohhira, T., Oshiro, E., Qi, D., Oshimura, M., Kugoh, H
  • Journal Title: Cytogenetic and Genome Research Volume: 125(1) Pages: 19-25
  • Peer Reviewed
• [Journal Article] Decreased PITX1 gene expression in human cutaneous malignant melanoma and its clinicopathological significance European
  • Author(s): Osaki M, Chinen H, Yoshida Y, Ohhira T, Sunamura N, Yamamoto O, Ito H, Oshimura M, Kugoh H
  • Journal Title: Journal of Dematology Volume: (in press)
  • Peer Reviewed
• [Presentation] hTERT発現を抑制するPitX1から解き明かすhTERT発現制御ネットワーク (2012)
  • Author(s): 吉川なつこ, 末松知久, 大平崇人, 押村光雄, 久郷裕之, 井上敏昭
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡, 福岡国際会議場・マリンメッセ福岡
  • Year and Date: 20121211-14
• [Presentation] 染色体免疫沈降法(ChrIP)を用いた疾患関連リピート配列に結合する因子の探索 (2012)
  • Author(s): 砂村直洋, 中山祐二, 荻野由加利, 難波栄二, 押村光雄, 久郷裕之
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡, 福岡国際会議場・マリンメッセ福岡
  • Year and Date: 20121211-14
• [Presentation] 新規テロメレース抑制遺伝子PITX1は悪性黒色腫の増殖を抑制する (2012)
  • Author(s): 大平崇人, 尾崎充彦, 砂村直洋, 吉田雄一, 山元 修, 押村光雄, 久郷裕之
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡, 福岡国際会議場・マリンメッセ福岡
  • Year and Date: 20121211-14
• [Presentation] PITX1発現低下は、ヒトメラノーマの進展に関与する (2012)
  • Author(s): 久郷裕之, 大平崇人, 砂村直洋, 井藤久雄, 押村光雄, 尾崎充彦
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: 札幌, ホテルロイトン札幌, さっぽろ芸文館, 札幌市教育文化会館
  • Year and Date: 20120919-21
• [Presentation] 低酸素によるhMSH3の発現抑制と大腸がんにおけるゲノム不安定性 (2012)
  • Author(s): 逸見 仁道, 有田 通恒, 小池 淳一, 菊池 由宣, 船橋 公彦, 大平 崇人, 久郷 裕之, 近藤 元就
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: 札幌, ホテルロイトン札幌, さっぽろ芸文館
  • Year and Date: 20120919-21
• [Presentation] Human artificial chromosomes for gene delivery and stable expression (2012)
  • Author(s): M. Oshimura, Y. Kazuki, M. Hiratsuka, T. Ohbayashi, Y. Iida, N. Uno, E. Ueno, K. Ueda, T. Ohhira, H. Kurosaki, S. Abe, H. Kugoh
  • Place of Presentation: PepTalk, Coronado, CA, USA, HotelDel Coronado
  • Year and Date: 20120109-13
• [Presentation] Characterization of a novel telomerase associated protein using microcell-mediated chromosome transfer (2012)
  • Author(s): H. Kugoh
  • Organizer: Interdisciplinary approaches for the study of senescence (UK/Japan Collaborative Symposium in association with The Japan Society for the Promotion of Science)
  • Place of Presentation: Cambridge, UK, Cancer Research UK, Cambridge Research Institute
  • Year and Date: 2012-02-09
• [Presentation] 特定染色体領域に集積する機能分子複合体の解析 (2011)
  • Author(s): 瀬古朋美, 山口繁幸, 吉村祐貴,中山祐二, 加藤基伸, 大林徹也, 押村光雄, 久郷裕之
  • Organizer: 第56回日本人類遺伝学会
  • Place of Presentation: 幕張, 幕張メッセ
  • Year and Date: 20111109-12
• [Presentation] 染色体導入による新規テロメラーゼ抑制遺伝子の同定 (2011)
  • Author(s): 久郷裕之, Dong-Lai Qi, 大平崇人, 太田力, 井上敏昭, 押村光雄
  • Organizer: 第56回日本人類遺伝学会
  • Place of Presentation: 幕張, 幕張メッセ
  • Year and Date: 20111109-12
• [Presentation] Identification of the chromatin regions coated by non-coding Xist RNA (2011)
  • Author(s): H. Kugoh, K. Murakami, E. Ohshiro, T. Ohhira and M. Oshimura
  • Organizer: 61^<th> American Society of Human Genetics (ASHG)
  • Place of Presentation: Montreal, Canada, MontrealConvention Centre
  • Year and Date: 20111011-15
• [Presentation] Evidence for a hTERT repressor gene on human chromosome 3p21.3 by using chromosome engineering (2011)
  • Author(s): T. Ohhira, S. Abe, H. Tanaka, T. Notsu, S. Horike, D.L. Qi, C. Fujisaki, D. Gilly, M. Oshimura, H. Kugoh
  • Organizer: 61^<th> American Society of Human Genetics (ASHG)
  • Place of Presentation: Montreal, Canada, Montreal Convention Centre
  • Year and Date: 20111011-15
• [Presentation] Construction of in vitro model system to understand molecular mechanism of CGG repeat expansion in Fragile X syndrome (2011)
  • Author(s): Y. Nakayama, T. Seko, N. Sunamura, Y. Ogino, M. Kato, E. Nanba, M. Oshimura, H. Kugoh
  • Organizer: 61^<th> American Society of Human Genetics (ASHG)
  • Place of Presentation: Montreal, Montreal Convention Centre, Canada
  • Year and Date: 20111011-15
• [Presentation] A novel proteomics approach for identification of protein-RNA complex associated with specific gene loci using a human artificial chromosome (HAC) (2011)
  • Author(s): T. Seko, S. Yamaguchi, Y. Yoshimura, Y. Nakayama, M. Kato, T. Obayashi, M. Oshimura, H. Kugoh
  • Organizer: 61^<th> American Society of Human Genetics (ASHG)
  • Place of Presentation: Montreal, Canada, Montreal Convention Centre
  • Year and Date: 20111011-15
• [Presentation] 染色体改変技術を応用したテロメレース抑制遺伝子領域 3p21.3の同定 (2011)
  • Author(s): 大平崇人,阿部智志,押村光雄,久郷裕之
  • Organizer: 第70回日本癌学会学術総会
  • Place of Presentation: 名古屋, 名古屋国際会議場
  • Year and Date: 20111003-05
• [Presentation] 標的ゲノム領域上の蛋白-RNA機能複合体解析(ChrIP法)に向けたヒト人工染色体(HAC)の活用 (2010)
  • Author(s): 瀬古朋美, 山口繁幸, 吉村祐貴, 青木慎介, 中山祐二, 加藤基伸, 大林徹也, 押村光雄, 久郷裕之
  • Organizer: 第33回日本分子生物学会年会
  • Place of Presentation: 神戸
  • Year and Date: 20101207-10
• [Presentation] 染色体改変技術を用いたヒト3p21.3領域内のTERT抑制遺伝子の同定 (2010)
  • Author(s): 大平崇人, 阿部智志, 田中宏美, 野津智美, 堀家慎一, 藤崎央子, Dong-Lai Qi, 押村光雄, 久郷裕之
  • Organizer: 第33回日本分子生物学会年会
  • Place of Presentation: 神戸
  • Year and Date: 20101207-10
• [Presentation] ヒト15番染色体を保持したトランスクロモソミックマウスの作出 (2010)
  • Author(s): 目黒-堀家牧子,宮野勝,杉原一司,成瀬智恵,久郷裕之,押村光雄,浅野雅秀,堀家慎一
  • Organizer: 第4回日本エピジェネティクス研究会年会
  • Place of Presentation: 米子
  • Year and Date: 20100528-29
• [Presentation] Localization of an hTERT repressor region on human chromosome 3p21.3 using chromosome engineering (2010)
  • Author(s): 大平崇人, 阿部智志, 田中宏美, 野津智美, 堀家慎一, 藤崎央子, Qi Dong-Lai, David Gilley, 押村光雄,久郷裕之
  • Organizer: がん若手研究者ワークショップ長野
  • Place of Presentation: アートランドホテル蓼科
  • Year and Date: 2010-09-03
• [Presentation] 機能性RNAによる染色体機能制御機構の解明 (2009)
  • Author(s): 久郷裕之
  • Organizer: 第60回染色体学会
  • Place of Presentation: 松江
  • Year and Date: 20091100
• [Presentation] Further investigation of IMMP2L (2009)
  • Author(s): N. H. Sykes, A. T. Pagnamenta, G. Lunter, H. Kugoh, M. Oshimura, Elena Bacchelli, Elena Maestrini, A. J. Bailey, A. P. Monaco
  • Organizer: LRRN3 and DOCK4 as potential candidate genes for autism susceptibility
  • Place of Presentation: ASHG, Hawaii
  • Year and Date: 20091020-24
• [Remarks] Nature日本語版FOCUS:長鎖ncRNAが操る「生命現象の裏社会」に.研究が紹介された。2011年12月
  • URL: http://www.natureasia.com/japan/nature/ad-focus/111201.php