1998 Fiscal Year Annual Research Report

サイクリン依存性キナーゼのターゲットとなる未知蛋白質の探索

Research Project

Project/Area Number	10480203
Research Category	Grant-in-Aid for Scientific Research (B)
Research Institution	National Cancer Center Research Institute and Research Center for Innovative Oncology, National Cancer Center Hospital East
Principal Investigator	田矢洋一国立がんセンター, 研究所生物学部, 室長 (60133641)
Keywords	サイクリンE / サイクリンD1 / Cdk4 / Cdk2 / NPAT
Research Abstract	NPAT遺伝子は、Ataxia telangiectasiaの原因遺伝子であるATM遺伝子にごく隣接して染色体上に存在している遺伝子として今井らによってクローニングされた。このNPAT蛋白質がCdk2-サイクリンEと強く結合して、しかも、そのキナーゼでリン酸化を受けることも昨年わかったので、NPAT上のCdk2-サイクリンE結合部位、および、リン酸化を受ける部位を同定する研究を進めた。その結果、2、3の候補の部位に絞り込むところまで進んだ。一方、RB蛋白質上にある約13ケ所のリン酸化部位を認識する抗体をほぼすべて作製したが、それを用いて、Cdk4によるリン酸化部位とCdk2によるリン酸化部位とを区別できることを示した。たとえば、Ser780、Ser795やSer608はCdk4によってかなり優先的にリン酸化されるが、逆に、Thr356、Ser612やThr821はCdk2かCdc2によってはよくリン酸化されるけれども、Cdk4によってはほとんどリン酸化されないことがわかった。さらに、これらを用いて、RB蛋白質のC末端側には、リン酸化に際してCdk2が先ず結合する部位が存在することも明らかにした。

Research Products
(11 results)

All Other

All Publications (11 results)

[Publications] Banin,S.et al.: "Enhanced phosphorylation of p53 by ATM in response to DNA damage." Science. 281. 1674-1677 (1998)
[Publications] Canman,C.E.et al.: "Activation of the ATM kinase by ionizing radiation and phosphorylation of p53." Science. 281. 1677-1679 (1998)
[Publications] Khanna,K.K.et al.: "ATM associates with and phosphorylates p53: mapping the region of interaction." Nature Genet.20. 398-400 (1998)
[Publications] Lu,H.et al.: "Ultraviolet radiation,but not gamma radiation or etoposide-induced DNA damage,results in the phosphorylation of the murine p53 protein at serine-389" Proc.Natl.Acad.Sci.USA. 95. 6399-6402 (1998)
[Publications] Tibbetts,R.S.et al.: "ATR is a DNA damage-responsive protein kinase that phosphorylates the p53 tumor suppressor protein." Genes & Dev.13. 152-157 (1999)
[Publications] Adams,P.D.et al.: "The retinoblastoma protein contains a C-terminal motif that targets it for phosphorylation by cyclin/cdk2 complexes." Mol.Cell.Biol.19. 1068-1080 (1999)
[Publications] Cuddihy,A.R.et al.: "The interferon-inducible protein kinase PKR mediates the transcriptional activation of the tumor suppressor p53." Mol.Cell.Biol.19. 2475-2484 (1999)
[Publications] Nakagawa,K.et al.: "Requirement of ATM in the phosphorylation of the human p53 protein at serine 15." Mol.Cell.Biol.19. 2828-2834 (1999)
[Publications] Brugarolas,J.et al.: "Inhibition of cyclin-dependent kinase 2 by p21 is necessary for retinoblastoma protein-mediated G1 arrest after g-irradiation." Proc.Natl.Acad.Sci.USA. 96. 1002-1007 (1999)
[Publications] Shieh,S.-Y.et al.: "DNA damage-inducible phosphorylation at N-terminal sites including a novel site,serine 20,requiresoliqomerization of p53." EMBO J.(in press). (1999)
[Publications] Nagata,Y.et al.: "The stabilization mechanism of mutant-type p53 by impaired ubiquitination: the loss of wild-type p53 function and the hsp90 association" Oncogene. (in press). (1999)

1998 Fiscal Year Annual Research Report

サイクリン依存性キナーゼのターゲットとなる未知蛋白質の探索

Principal Investigator

田矢 洋一 国立がんセンター, 研究所生物学部, 室長 (60133641)

Research Products

[Publications] Banin,S.et al.: "Enhanced phosphorylation of p53 by ATM in response to DNA damage." Science. 281. 1674-1677 (1998)

[Publications] Canman,C.E.et al.: "Activation of the ATM kinase by ionizing radiation and phosphorylation of p53." Science. 281. 1677-1679 (1998)

[Publications] Khanna,K.K.et al.: "ATM associates with and phosphorylates p53: mapping the region of interaction." Nature Genet.20. 398-400 (1998)

[Publications] Lu,H.et al.: "Ultraviolet radiation,but not gamma radiation or etoposide-induced DNA damage,results in the phosphorylation of the murine p53 protein at serine-389" Proc.Natl.Acad.Sci.USA. 95. 6399-6402 (1998)

[Publications] Tibbetts,R.S.et al.: "ATR is a DNA damage-responsive protein kinase that phosphorylates the p53 tumor suppressor protein." Genes & Dev.13. 152-157 (1999)

[Publications] Adams,P.D.et al.: "The retinoblastoma protein contains a C-terminal motif that targets it for phosphorylation by cyclin/cdk2 complexes." Mol.Cell.Biol.19. 1068-1080 (1999)

[Publications] Cuddihy,A.R.et al.: "The interferon-inducible protein kinase PKR mediates the transcriptional activation of the tumor suppressor p53." Mol.Cell.Biol.19. 2475-2484 (1999)

[Publications] Nakagawa,K.et al.: "Requirement of ATM in the phosphorylation of the human p53 protein at serine 15." Mol.Cell.Biol.19. 2828-2834 (1999)

[Publications] Brugarolas,J.et al.: "Inhibition of cyclin-dependent kinase 2 by p21 is necessary for retinoblastoma protein-mediated G1 arrest after g-irradiation." Proc.Natl.Acad.Sci.USA. 96. 1002-1007 (1999)

[Publications] Shieh,S.-Y.et al.: "DNA damage-inducible phosphorylation at N-terminal sites including a novel site,serine 20,requiresoliqomerization of p53." EMBO J.(in press). (1999)

[Publications] Nagata,Y.et al.: "The stabilization mechanism of mutant-type p53 by impaired ubiquitination: the loss of wild-type p53 function and the hsp90 association" Oncogene. (in press). (1999)

田矢洋一国立がんセンター, 研究所生物学部, 室長 (60133641)