2000 Fiscal Year Annual Research Report

NMDA受容体と癌抑制遺伝子産物APCおよびDLGファミリーの複合体形成の意義

Research Project

Project/Area Number	11470039
Research Institution	The University of Tokyo
Principal Investigator	秋山徹東京大学, 分子細胞生物学研究所, 教授 (70150745)
Keywords	NMDA受容体 / APC / PSD-95 / SPAL / 記憶 / 長期増強 / シナプス可塑性 / 癌抑制遺伝子
Research Abstract	シナプス可塑性に関わるNMDA受容体の調節、シグナル伝達機構の理解には、NMDA受容体と複合体を形成する蛋白質の同定と機能の解析が極めて重要であると考えられる。我々は、NMDA受容体がDLGファミリーの蛋白質PSD-95を介して大腸癌の癌抑制遺伝子APCの産物をはじめDAP、新規GAP(SPALと命名)、LGN、Fynなど多数の分子と複合体を形成していることを見出し、これらの蛋白質の機能を解析した。 1)APC-PSD-95-NMDA受容体複合体はAPCが結合したPSD-95とNMDA受容体が結合したPSD-95が多量体をつくることにより形成されると考えられた。PSD-95との結合部位の欠失したAPCを発現するマウスは体が小さく、運動能および日周リズムの軽度の異常がみられた。water mazeの実験からは目立った学習能の低下は見出されなかった。 2)PSD-95のguanylate kinaseドメインに結合するSPALは、GAPドメイン、PDZドメイン、leucine zipperドメインをもつ分子量約200kDaの蛋白質で、Rapl特異的なGAP活性をもち、特に海馬、嗅球、大脳皮質などで強く発現していた。また、培養海馬神経細胞では細胞体に加えてシナプスに濃縮していることが観察された。さらに免疫沈降実験によりSPALはNMDA受容体-PSD-95複合体に含まれていることが明らかになった。NMDA受容体を介したシグナル伝達に関与する可能性を検討したところ、培養海馬神経細胞にNMDAを作用させた時に脱リン酸化をおこすこと、活性化がおきることが見出された。さらにSPALの機能を明らかにするためにSPAL遺伝子破壊マウスの作製を進めた。また、SPALのPDZドメインおよびleucine zipperに結合する蛋白質を見出し解析を進めた。

Research Products

(19 results)

All Other

All Publications (19 results)

[Publications] F.Hamada: "Negative regulation of Wingless signaling by D-Axin, a Droso phila homologue of Axin."Science. 283. 1739-1742 (1999)
[Publications] T.Yamochi: "Adenovirus-mediated high expression of BCL-6 in CV-1 cells induces apoptotic cell death accompanied by downregulation of BCL-2 and BCL-XL."Oncogene. 18. 487-494 (1999)
[Publications] T.Tezuka: "PSD-95 promoles Fyn-mediated tyrosine phosphorylation of the N-methyl-D-aspartate receptor subunil NR2A."Proc.Nall.Acad.Sci.USA. 96. 435-440 (1999)
[Publications] T.Senda: "Localization of MCC (mutated in colorectal cancer) in various tissues of mice and its involvement in cell differentiation."J.Histochem Cytochem. 47. 1149-1158 (1999)
[Publications] C.R,Badal: "Localization of the GAP family SPAL in the Rat Esophagus and Heart."Med.Elcetron Microse.. 32. 20-24 (1999)
[Publications] T.Suzuki: "Tax oncoprotein of HTLV-1 binds to the human homologue of Drosophila dises large tumor suppressor protein. hDLG. and perturbs its function in cell growth control."Oncogene.. 18. 5967-5972 (1999)
[Publications] Y.Kawasaki: "Asef, a Link between the Tumor Suppressor APC and G-protain Signaling."Science. 289. 1194-1197 (2000)
[Publications] H.Haraguchi: "The hDLG-associated protein DAP interacts with dynein light chain and neuronal nitric oxide synthase."Gene cells. 5. 905-911 (2000)
[Publications] Y.Dabashi: "APC protein is required for initiation of neuronal differentiation in rat pheochromocytoma PC12 cells."Biochem.Biopys.Res.Commun.. 279. 685-691 (2000)
[Publications] T.Senda: "Expression of APC in the mouse kideney and its colocalization with A-canenin and hDLG."J.Histochem.Cytochem.. (in press).
[Publications] Kohu,K.: "Localization of the Novel GAP Family Protein SPAL in Epithelial cells."Acta Histochem.Cytochem.. (in press).
[Publications] Kakinuma,N: "APC is Colocalized with β-catenin and hDLG in the Henle's Loop of the Mouse kidney."Acta Histochem.Cytochem.. (in press).
[Publications] T.Ishidate: "The APC-hDLG Complex Negatively Regulates Cell Cycle Progression from the G0/G1 to Sphase."Oncogene. 19. 365-372 (2000)
[Publications] H.Nakagawa: "EB3, a novel member of the EB1 family preferentially expressed in the central nervous system, bins to a CNS-specific."Oncogene. 19. 210-216 (2000)
[Publications] H.Nakagawa: "APCL, a central nervous system-specific homolog of adenomatous polysois coli tumor suppressor, binds to p53-binding protein 2 and translocates."Cancer Res.. 60. 101-105 (2000)
[Publications] K.Kawahara: "Downregulation of b-catenin by the Colorectal Tumor Suppressor APC Requires Association with Axin and β-catenin."J.Biol.Chem. 275. 8369-8374 (2000)
[Publications] T.Akiyama: "Wht/β-catenin signaling."Cytokine & Growth Factor Reviews. 11. 273-282 (2000)
[Publications] H.Yanai: "The Colorectal Tumor Suppressor APC is Present in the NMDA-receptor-PDS-95 Complex in the Brain."Genes Cells. 5. 815-822 (2000)
[Publications] Tago,K: "Inhibition of Wnt signaling by ICAT, a Novel b-catenin-interacting Protein."Gense Dev. 14. 1741-1749 (2000)

2000 Fiscal Year Annual Research Report

NMDA受容体と癌抑制遺伝子産物APCおよびDLGファミリーの複合体形成の意義

Principal Investigator

秋山 徹 東京大学, 分子細胞生物学研究所, 教授 (70150745)

Research Products

[Publications] F.Hamada: "Negative regulation of Wingless signaling by D-Axin, a Droso phila homologue of Axin."Science. 283. 1739-1742 (1999)

[Publications] T.Yamochi: "Adenovirus-mediated high expression of BCL-6 in CV-1 cells induces apoptotic cell death accompanied by downregulation of BCL-2 and BCL-XL."Oncogene. 18. 487-494 (1999)

[Publications] T.Tezuka: "PSD-95 promoles Fyn-mediated tyrosine phosphorylation of the N-methyl-D-aspartate receptor subunil NR2A."Proc.Nall.Acad.Sci.USA. 96. 435-440 (1999)

[Publications] T.Senda: "Localization of MCC (mutated in colorectal cancer) in various tissues of mice and its involvement in cell differentiation."J.Histochem Cytochem. 47. 1149-1158 (1999)

[Publications] C.R,Badal: "Localization of the GAP family SPAL in the Rat Esophagus and Heart."Med.Elcetron Microse.. 32. 20-24 (1999)

[Publications] T.Suzuki: "Tax oncoprotein of HTLV-1 binds to the human homologue of Drosophila dises large tumor suppressor protein. hDLG. and perturbs its function in cell growth control."Oncogene.. 18. 5967-5972 (1999)

[Publications] Y.Kawasaki: "Asef, a Link between the Tumor Suppressor APC and G-protain Signaling."Science. 289. 1194-1197 (2000)

[Publications] H.Haraguchi: "The hDLG-associated protein DAP interacts with dynein light chain and neuronal nitric oxide synthase."Gene cells. 5. 905-911 (2000)

[Publications] Y.Dabashi: "APC protein is required for initiation of neuronal differentiation in rat pheochromocytoma PC12 cells."Biochem.Biopys.Res.Commun.. 279. 685-691 (2000)

[Publications] T.Senda: "Expression of APC in the mouse kideney and its colocalization with A-canenin and hDLG."J.Histochem.Cytochem.. (in press).

[Publications] Kohu,K.: "Localization of the Novel GAP Family Protein SPAL in Epithelial cells."Acta Histochem.Cytochem.. (in press).

[Publications] Kakinuma,N: "APC is Colocalized with β-catenin and hDLG in the Henle's Loop of the Mouse kidney."Acta Histochem.Cytochem.. (in press).

[Publications] T.Ishidate: "The APC-hDLG Complex Negatively Regulates Cell Cycle Progression from the G0/G1 to Sphase."Oncogene. 19. 365-372 (2000)

[Publications] H.Nakagawa: "EB3, a novel member of the EB1 family preferentially expressed in the central nervous system, bins to a CNS-specific."Oncogene. 19. 210-216 (2000)

[Publications] H.Nakagawa: "APCL, a central nervous system-specific homolog of adenomatous polysois coli tumor suppressor, binds to p53-binding protein 2 and translocates."Cancer Res.. 60. 101-105 (2000)

[Publications] K.Kawahara: "Downregulation of b-catenin by the Colorectal Tumor Suppressor APC Requires Association with Axin and β-catenin."J.Biol.Chem. 275. 8369-8374 (2000)

[Publications] T.Akiyama: "Wht/β-catenin signaling."Cytokine & Growth Factor Reviews. 11. 273-282 (2000)

[Publications] H.Yanai: "The Colorectal Tumor Suppressor APC is Present in the NMDA-receptor-PDS-95 Complex in the Brain."Genes Cells. 5. 815-822 (2000)

[Publications] Tago,K: "Inhibition of Wnt signaling by ICAT, a Novel b-catenin-interacting Protein."Gense Dev. 14. 1741-1749 (2000)

秋山徹東京大学, 分子細胞生物学研究所, 教授 (70150745)