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1994 Fiscal Year Annual Research Report

癌抑制遺伝子産物の生理機能

Research Project

Project/Area Number 06280124
Research InstitutionNational 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, 生物学部, 室長 (60133641)

Co-Investigator(Kenkyū-buntansha) 原田 久士  阪大, 細胞生体工学センター, 助手 (10222233)
松七五三 仁  東大, 医科研, 助教授 (50251442)
野田 哲生  癌研究所, 部長 (10183550)
土田 信夫  東京医科歯科大, 歯, 教授 (60089951)
小田 鈎一郎  東京理科大, 基礎工, 教授 (40012736)
KeywordsRB蛋白質 / p53 / 癌抑制遺伝子 / サイクリンD / Cdk4 / Cdkインヒビター / mgl-1
Research Abstract

RB蛋白質をリン酸化するRBキナーゼについては、サイクリンD1-Cdk4が、他のサイクリンEやA依存性キナーゼとは異なったアミノ酸配列の所をリン酸化することを初めて明らかにした。さらに、RB蛋白質上にはサイクリンD1-Cdk4でリン酸化されるけれども他のサイクリンEやA依存性キナーゼではリン酸化されない部位が存在することも見い出した。サイクリンD1は癌遺伝子にもなるサイクリンであるので、この発見によって、サイクリンD1が細胞癌化に関与するメカニズムの解明が大きく進展すると期待できる。一方では、以前に開発したリン酸化ペプチドの化学合成法を用いて、RB蛋白質上の3ヵ所のリン酸化部位をそれぞれ認識する抗体の作製に成功した。これには、上述のサイクリンD1-Cdk4特異的なリン酸化部位も含まれる。
また、接触阻止においてはサイクリンD依存性キナーゼ活性が阻害されるが、その際、サイクリンDとCdk4は複合体を形成しているのに、Cdk4の活性化には必須であるThr^<172>がリン酸化されていないことを見い出した。しかし、この部位をリン酸化する酵素であるCdk活性化キナーゼ(CAK)の活性には大きな変化はなかった。一方、_p27^<kip1>が多量に存在することで、CAKがCdk4を基質として認識するのを阻害し、結果として、接触阻止ではサイクリンD依存性キナーゼの活性化が阻害されていることを明らかにした。
p53については、ヒトのp53温度感受性変異株(バリン138)を作製した。この変異株をラット胎児繊維芽細胞に活性化rasとともに導入した細胞では、waf1発現の上昇を経ずにG1停止が起きること、Saos-2に導入した細胞ではG1とG2/Mの2つの時期で停止が起きることが示唆された。
また、ショウジョウバエの癌抑制遺伝子の1(2)glのマウスホモログmgl-1遺伝子の機能をジーンターゲッティングにより解析した結果、mgl-1を欠損したマウスでは胎生期の神経上皮細胞の異常増殖が起きることが判明したが、腫瘍の発生は見られなかった。

  • Research Products

    (40 results)

All Other

All Publications (40 results)

  • [Publications] Toshinori Ozaki et al.: "Complex formation between lamin A and the retinoblastoma gene product:Identification of the domain on lamin A required for its interaction." Oncogene. 9. 2649-2653 (1994)

  • [Publications] Kitagawa,M.et al.: "A cyclin-dependent kinase inhibitor,butyrolactone I,inhibits phosphorylation of RB protein and cell cycle progression." Oncogene. 9. 2549-2557 (1994)

  • [Publications] Saijo,M.et al.: "Inactivation of oncoprotein binding by a single Cys^<706> to Tyr substitution in the retinoblastoma protein." FEBS Lett.340. 181-184 (1994)

  • [Publications] Kitagawa,M.et al.: "Phosphorylation of E2F-1 by cyclin A-cdk2." Oncogene. 10. 229-236 (1995)

  • [Publications] Ueno,Y.et al.: "Chemical synthesis of phosphopeptides using arylthio group for protection of phosphate:Application to identification of cdc2 kinase-phosphorylation sites." Int.J.Peptide Prot.Res.(in press). (1995)

  • [Publications] Ueno,Y.et al.: "Solid-phase synthesis of peptides containing ο-phosphoryl serine and ο-phosphoryl threonine using allyl group for phosphate protection." Bioorg.Med,Chem.Lett.(in press). (1995)

  • [Publications] Suzuki-Takahashi,I.et al.: "Interaction of E2F with pRB and p107 are regulated through phosphorylation of pRB and p107 by cyclin-dependent kinase." Oncogene. (in press). (1995)

  • [Publications] Hiwasa,T.et al.: "Inhibition of cysteine proteinases and proteosome by Ras,Ran and cyclin." Proteases and Cancer(Monduzzi Editore,Italy). (in press). (1995)

  • [Publications] Saijo,M.et al.: "Molecular cloning of a human protein which binds to the retinoblastoma protein and chromosomal mapping." Genomics. (in press). (1995)

  • [Publications] Taya,Y.: "Cell cycle-dependent phosphorylation of the tumor suppressor RB protein" Molecules and Cells. (in press). (1995)

  • [Publications] Matsushime,H.et al.: "D-type cyclin-dependent kinase activity in mammalian cells." Mol.Cell.Biol.14. 2066-2076 (1994)

  • [Publications] Nakajima,T.et al.: "Adenovirus EIA-induced apoptosis elicits a steep decrease in the topoisomerase IIα level during the latent phase." Oncogene. (in press). (1995)

  • [Publications] Nakamura,T.et al: "Down-regulation of the cyclin A promoter in differentiating human embryonal carcinoma cells is mediated by the depletion of ATF-1 and ATF-2 in the complex at the ATF/CRE site." Exp.Cell Res.216(in press). (1995)

  • [Publications] Icho,T.et al.: "A novel human gene that is preferentially in heart muscle." Gene. 144. 301-306 (1994)

  • [Publications] Tsutsumi-Ishii,Y.et al: "Response of heat shock element within the human HSP70 promoter to mutated p53 genes." Cell Growth Diff.6. 1-8 (1995)

  • [Publications] Hirano,Y.et al.: "Role of the p53 mutation in cell line estsblishment and identification of the minimum trans-activation and transform suppression domains." Eur.J.Cancer B.Oral Oncol.(in press). (1995)

  • [Publications] Hirano,Y.et al.: "A temperature sensitive mutaint of the human p53,Val138,arrests rat cell growth without induced expression of cip1/waf1/sdi1 after temperature shift down." Oncogene. (in press). (1995)

  • [Publications] Sugito,K.et al.: "Interaction between hsp70 and hap40,eukaryotic homologues of DnaK and DnaJ,in human cells expressing mutant-type p53." FEBS Lett.358. 161-164 (1995)

  • [Publications] Sekiguchi,T.et al.: "Apoptosis is induced in BHK cells by the tsBN462/13 mutation in the CCG1/TAFII250 subunit of the TFIID basal transcription factor." Exp.Cell Res.(in press). (1995)

  • [Publications] Harada,A.et al.: "Altered microtuble organization in small-calibre axons of mice lacking tau protein." Nature. 369. 488-491 (1994)

  • [Publications] Takeshima,H.et al.: "Excitation-contraction uncoupling and muscular degeneration in mice lacking functional skeltal muscle ryanodine-receptor gene." Nature. 369. 555-559 (1994)

  • [Publications] Hirai,Y.et al.: "Establishment and characterization of adenocarcinoma cell lines derived from endometrial serous papillary adenocarcinoma." Gynecologic Oncol.54. 184-195 (1994)

  • [Publications] Shiba,K.et al.: "Human cytoplasmic isoleucy1-tRNA synthetase:Selective divergence of the anticodon-binding domain and acquisition of a new structural unit." Proc.Natl.Acad.Sci.USA. 91. 7435-7439 (1994)

  • [Publications] Shiba,K.et al.: "Human glycyl tRNA-synthetase:Wide divergence of primary structure from bacterial counterpart and species-specific aminoacylation." J.Biol.Chem. 269. 30049-30055 (1994)

  • [Publications] Kamada,S.et al.: "bcl-2 deficiency in mice leads to pleiotropic abnormalities:Accelerated lymphoid cell death in thymus and spleen,polycystic kidney,hair hypopigmentation,and distoted small intestine." Cancer Res.55. 354-359 (1995)

  • [Publications] Matsuhisa,A.et al.: "Inositol monophosphatase activity from the Escherichia coli suhB gene product." J.Bacteriol.177. 200-205 (1995)

  • [Publications] Uehara,Y.et al.: "Placental defect and embryonic lethality in mice lacking hepatocyte growth factor/scatter factor." Nature. 373. 702-705 (1995)

  • [Publications] Kanda,T.et al.: "Wild type p53 Transactivate both transcription and DNA replication through the DNA binding site." Mol.Cell.Biol.14. 2651-2663 (1994)

  • [Publications] Ushijima,T.et al.: "Presence of p53 mutation in 3Y1-B clones 1-6:a rat cell line widely used as a normal immortalized fibroblast." Jpn.J.Cancer Res.85. 455-458 (1994)

  • [Publications] Yamamoto,H.et al.: "The oncogenic transcription factor IRF-2 possesesa transcriptional repression and a latent activation domain." Oncogene. 9. 1423-1428 (1994)

  • [Publications] Tanaka,N.,et al.: "Cellular commitment to oncogene-induced transformation or apoptosis is dependent on the transcription factor IRF-1." Cell. 77. 829-839 (1994)

  • [Publications] Kimura,T.et al.: "Involvement of the IRF-1 transcription factor in antiviral responses to interferons." Science. 264. 1921-1924 (1994)

  • [Publications] Harada,H.et al.: "Accelerated exon skipping of IRF-1 mRNA in human myelodysplasia/leukemia;a possible mechanism of tumor supressor inactivation." Oncogene. 9. 3313-3320 (1994)

  • [Publications] Sawada,Y.et al.: "Identification of functional domains of adenovirus tumor specific transplantation antigen in type 5 and 12 by viable viruses carrying chimeric E1A genes." Int.J.Cancer. 57. 598-603 (1994)

  • [Publications] Takimoto,M.et al.: "Identification of cellular proteins that bind the central conserved region of p53." Biochem.Biophys.Res.Commun.202. 490-496 (1994)

  • [Publications] Shibuya,H.et al.: "Functional dissection of p56^<1ck>,a protein tyrosine kinase which mediates interleukine-2-induced activation of the c-fos gene." Mol.Cell.Biol.14. 5812-5819 (1994)

  • [Publications] Inoue,T.et al.: "Transduction of BCR-ABL cDNA causes alteration in factor-dependence and interferon response of human factor-dependent TF-1 cells." Blood. 84. 140a (1994)

  • [Publications] Shoji,W.et al.: "The helix-loop-helix protein Id inhibits differentiation of murine erythroleukemia cells." J.Biol.Chem.269. 5078-5084 (1994)

  • [Publications] Saito,F.et al.: "Direct mapping of the human TATA box-binding protein(TBP)gene to 6q27 by fluorescence in situ hybridization." Jpn.J.Human Genet.39. 421-425 (1994)

  • [Publications] Takiguchi,M.et al.: "The role of the conserved residue in pocket A and the polymorphic residue in pocket E of HLA-B^*3501 in presentaion of human minor histocompatibility peptides to T cells." Internat.Immunol.6. 1345-1352 (1994)

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Published: 1996-04-08   Modified: 2016-04-21  

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