1998 Fiscal Year Annual Research Report

リン酸化によるRB蛋白質とp53の生理機能の制御

Research Project

Project/Area Number	09254271
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	RB蛋白質 / サイクリンD1 / サイクリンE / Cdk4 / Cdk2 / p53 / ATM / ATR
Research Abstract	1. p53上の推定されるリン酸化部位(約13カ所)を特異的に認識する抗体をほぼすべて作製した。C末端側の2カ所のアセチル化部位に対する抗体も作製した。 2. Ataxia telangiectaciaの原因遺伝子であるATM蛋白質がプロテインキナーゼの活性をもち、しかも、p53のSer15を直接にリン酸化することを証明した。 3. ATMのファミリーであるATRについても同様のことを見い出した。 4. DNAダメージ後に、p53のSer20,46のリン酸化やLys373,382のアセチル化が起きることも見い出した。 5. N末端側のSer15,20,33がin vivoでリン酸化されるためには、C末端側にあるテトラマー形成ドメインが必要であることを見つけた。 6. 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。 7. 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

リン酸化によるRB蛋白質とp53の生理機能の制御

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)