2003 Fiscal Year Annual Research Report

霊長類ES細胞維持因子の同定とその機能解析

Research Project

Project/Area Number	15039207
Research Institution	The University of Tokyo
Principal Investigator	北村俊雄東京大学, 医科学研究所, 教授 (20282527)
Co-Investigator(Kenkyū-buntansha)	杉山拓也東京大学, 医科学研究所, 日本学術振興会特別研究員
Keywords	発現クローニング / ES細胞 / サル / LIF / 自己複製
Research Abstract	哺乳動物細胞を利用した発現クローニング法により、霊長類のES細胞の維持に必要なマウス胎児性維持芽細胞(MEF)由来の因子の同定を、カニクイザルのES細胞を標的細胞として試みた。現在までにMEF細胞由来のライブラリー5万クローンをスクリーニングしたが、ES細胞の未熟状態を維持しうる因子は同定できていない。現在、スクリーニングを継続中である。またMEFで発現しているいくつかの分泌蛋白質のES細胞維持活性を調べることにより、そのうちWnt3にES細胞維持活性を有することが判明した。この活性はWnt3の可溶型レセプターにより中和されることから、Wnt3のレセプターを介したものであることが確認された。マウスES細胞においても同様の結果を確認した。これらの結果はWnt3Aが造血幹細胞だけでなくES細胞の未熟性を維持する活性も有する重要な因子であることを示している。ただし、Wnt3AのみではLIFのようにES細胞を長期維持することは難しい。つまり、マウスではLIFのみでES細胞の未熟性を保ったまま長期間培養できるが、サルES細胞はWnt3Aで長期間未熟性を維持することは不可能である。このことは、MEF由来でWnt3A以外にES細胞の未熟性維持に必要な因子が存在することを示唆している。この分子がWnt3Aと共同して働くのか、あるいは別に働くのかはこれまでの実験結果では明確ではないが、現在Wnt3Aの存在下においてMEF由来分子をスクリーニングする発現クローニング法の基礎検討を行っている。

Research Products

(12 results)

All Other

All Publications (12 results)

[Publications] Yujiri, T.: "MEK kinase 1 interacts with focal adhesion kinase and regulates insulin receptor Substrate-1 expression."The Journal of Biological Chemistry. 278. 3846-3851 (2003)
[Publications] Ozawa, T.: "A genetic approach to identifying mitochondrial proteins."Nature Biotechnology. 21. 287-293 (2003)
[Publications] Minoshima, Y.: "Phosphorylation by Aurora B converts MgcRacGAP to a RhoGAP during cytokinesis."Developmental Cell. 4. 549-560 (2003)
[Publications] Nosaka, T.: "Mammalian twisted gastulation is essential for skeleto-lymphogenesis."Molecular and Cellular Biology. 23. 2969-2980 (2003)
[Publications] Kitamura, T.: "Retrovirus-mediated gene transfer and expression cloning, powerful tools in functional genomics."Experimental Hematology. 31. 1007-1014 (2003)
[Publications] Bao, Y-C.: "Identification of a human cDNA sequence which encodes a novel membrane-associated protein containing a zinc metalloprotease motif."DNA Research. 10. 123-128 (2003)
[Publications] Murata, K.: "A novel tyrosine kinase inhibitor GTP14564 specifically inhibits constitutively active fms-like tyrosine kinase 3 through inactivation of STAT5."The Journal of Biological Chemistry. 278. 32892-32898 (2003)
[Publications] Ueno, H.: "A stromal cell-derived membrane protein that supports hematopoietic stem cells."Nature Immunology. 4. 457-463 (2003)
[Publications] Hisaoka, T.: "Expression of a member of tumor necrosis factor receptor superfamily, TROY, in the developing and the adult mouse brain."Brain Research. 143. 105-109 (2003)
[Publications] Takizawa, M.: "Requirement of gp130 signaling for the AGM hematopoiesis."Experimental Hematology. 31. 283-289 (2003)
[Publications] Tahara, H.: "Reconstitution of CD8+T cells by retroviral transfer of the T-cell receptor ab chain genes isolated from a clonally expanded P815 infiltrating lymphocytes."The Journal of Immunology. 171. 2154-2160 (2003)
[Publications] Kumagai, H.: "Identification and characterization of a new pair of immunoglobulin-like receptors LMIR1 and 2 derived from murine bone marrow-derived mast cells."Biochemical and Biophysical Research Communications. 307. 719-729 (2003)

2003 Fiscal Year Annual Research Report

霊長類ES細胞維持因子の同定とその機能解析

Principal Investigator

北村 俊雄 東京大学, 医科学研究所, 教授 (20282527)

Research Products

[Publications] Yujiri, T.: "MEK kinase 1 interacts with focal adhesion kinase and regulates insulin receptor Substrate-1 expression."The Journal of Biological Chemistry. 278. 3846-3851 (2003)

[Publications] Ozawa, T.: "A genetic approach to identifying mitochondrial proteins."Nature Biotechnology. 21. 287-293 (2003)

[Publications] Minoshima, Y.: "Phosphorylation by Aurora B converts MgcRacGAP to a RhoGAP during cytokinesis."Developmental Cell. 4. 549-560 (2003)

[Publications] Nosaka, T.: "Mammalian twisted gastulation is essential for skeleto-lymphogenesis."Molecular and Cellular Biology. 23. 2969-2980 (2003)

[Publications] Kitamura, T.: "Retrovirus-mediated gene transfer and expression cloning, powerful tools in functional genomics."Experimental Hematology. 31. 1007-1014 (2003)

[Publications] Bao, Y-C.: "Identification of a human cDNA sequence which encodes a novel membrane-associated protein containing a zinc metalloprotease motif."DNA Research. 10. 123-128 (2003)

[Publications] Murata, K.: "A novel tyrosine kinase inhibitor GTP14564 specifically inhibits constitutively active fms-like tyrosine kinase 3 through inactivation of STAT5."The Journal of Biological Chemistry. 278. 32892-32898 (2003)

[Publications] Ueno, H.: "A stromal cell-derived membrane protein that supports hematopoietic stem cells."Nature Immunology. 4. 457-463 (2003)

[Publications] Hisaoka, T.: "Expression of a member of tumor necrosis factor receptor superfamily, TROY, in the developing and the adult mouse brain."Brain Research. 143. 105-109 (2003)

[Publications] Takizawa, M.: "Requirement of gp130 signaling for the AGM hematopoiesis."Experimental Hematology. 31. 283-289 (2003)

[Publications] Tahara, H.: "Reconstitution of CD8+T cells by retroviral transfer of the T-cell receptor ab chain genes isolated from a clonally expanded P815 infiltrating lymphocytes."The Journal of Immunology. 171. 2154-2160 (2003)

[Publications] Kumagai, H.: "Identification and characterization of a new pair of immunoglobulin-like receptors LMIR1 and 2 derived from murine bone marrow-derived mast cells."Biochemical and Biophysical Research Communications. 307. 719-729 (2003)

北村俊雄東京大学, 医科学研究所, 教授 (20282527)