1995 年度実績報告書

癌抑制遺伝子産物の生理機能

研究課題

研究課題/領域番号	07272110
研究機関	国立がんセンター
研究代表者	田矢洋一国立がんセンター, 研究所生物学部, 室長 (60133641)
研究分担者	神田忠仁国立予防衛生研究所, 室長 (60134615) 松七五三仁東京大学, 医科学研究所, 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学, 歯学部, 教授 (60089951) 小田鈎一郎東京理科大学, 基礎工学部, 教授 (40012736)
キーワード	RB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 / ヒトパピローマウィルス
研究概要	1.酵母のTwo-hybrid系を利用して、HPV16のE6蛋白と複合体を形成する2つの細胞蛋白を得た。1つは、HUMHMCM2(酵母MCM2のヒトホモログ)であり、他の1つは未知の蛋白(約60Kd)であった。 2.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。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。 3.Drosophilaの癌抑制遺伝子l(2)glのマウスホモログmgl-1ならびにmgl-2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。mgl-1ホモ欠損マウスでは水頭症を呈し、出生後48時間以内に死亡し、組織学的には神経上皮細胞の異常が認められた。一方、mgl-2ホモ欠損マウスは、ほとんどが生殖能力を有する生体となるが、すべて矮小であった。 4.p21の変異蛋白を複数作製して精製し、アミノ酸置換されたp21はサイクリンDl/Cdk4複合体に結合するがその酵素活性を阻害できないことを見い出した。この変異p21をp16とともにヒト細胞に発現させると、p16に拮抗的な変異p21の作用が観察された。 5.ラットcdc2プロモーターのGl/S境界での活性化に関与するエンハンサーAAGTTACAAATAに結合する蛋白因子cdc2E1とcdc2E2のcDNAを単離した結果、E1はSRE中のCArGbox結合因子をコードし、E2は未知の遺伝子であることがわかった。

研究成果
(46件)

すべてその他

すべて文献書誌 (46件)

[文献書誌] Kitagawa, M. et al.: "Phosphorylation of E2F-1 by cyclin A-cdk2." Oncogene. 10. 229-236 (1995)
[文献書誌] 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.46. 106-112 (1995)
[文献書誌] Hiwasa, T. et al.: "Inhibition of cysteine proteinases and proteosome by Ras, Ran and cyclin." Proteases Involved in Cancer. 183-187 (1995)
[文献書誌] Suzuki-Takashi, I. et al.: "The interaction of E2F with pRB and p107 are regulated via the phosphorylation of pRB and p107 by cyclin-dependent kinase." Oncogene. 10. 1691-1698 (1995)
[文献書誌] Ueno, Y., et al.: "Solid-phase synthesis of peptides containing O-phosphoryl serine and O-phosphoryl threonine using allyl group for phosphate protection." Bioorg. Med. Chem. Let.5. 823-826 (1995)
[文献書誌] Saijo, M. et al.: "Molecular cloning of a human protein that binds to the retinoblastoma protein and chromosomal mapping." Genomics. 27. 551-519 (1995)
[文献書誌] Taya, Y.: "Cell cycle-dependent phosphorylation of the tumor suppressor RB protein." Molecules and Cells. 5. 191-195 (1995)
[文献書誌] Inoue, A. et al.: "70-kDa heat shock cognate protein directly interacts with N-terminal region of the retinoblastoma gene product pRb-identification of a novel region of pRb mediating protein interaction." J. Biol. Chem.270. 22571-22576 (1995)
[文献書誌] Sakai, Y. et al.: "cDNA sequence, chromosomal localization of a novel human protein RBQ-1 which binds to the retinoblastoma gene product." Genomics. 30. 98-101 (1995)
[文献書誌] Higashi, H. et al.: "Differences in substrate specificity between Cdk2-cyclin A and Cdk2-cyclin E in vitro." Biochem. Biophys. Res. Commun.216. 520-525 (1995)
[文献書誌] Higashi, H. et al.: "Cdk2 forms an inactive complex with cyclin D1 since Cdk2 associated with cyclin D1 is not phosphorylated by Cdk7-cyclin H." Eur. J. Biochem.(in press). (1996)
[文献書誌] Kitagawa, M. et al: "The consensus motif for phosphorylation by cyclin D1-Cdk4 is different from that of phosphorylation by cyclin A/E-Cdk2." EMBO J.(in press).
[文献書誌] Suziki, M. et al.: "An E1A-inducible Sp 1 negative regulator, G10BP represses transcription of the rat fibronectin gene." Mol. Cell. Biol.15. 5423-5433 (1995)
[文献書誌] Shimizu, M. et al.: "The G1/S boundary-specific enhancer of the rat cdc2 promoter." Mol. Cell. Biol.15. 2882-2892 (1995)
[文献書誌] Nakajima, T. et al.: "Adenovirus E1A-induced apoptosis elicits a steep decrease in the topoisomerase II a level during the latent phase." Oncogene. 10. 651-662 (1995)
[文献書誌] Nakamra, T. et al.: "Down-regulation of the cyclin A-promoter in differentiating human embryonal carcinoma cells is mediated by depletion of ATF-1 and ATF-2 in the complex at the ATF/CRE site." Exp. Cell. Res.216. 422-430 (1995)
[文献書誌] Kamada, S. et al.: "bcl-2 Deficiency in mice leads to pleiotropic abnormalities : A ccelerated lymphoid cell death in thymus and spleen, polycystic kidney, hair hypopigmentation, and distorted small intestine." Cancer Res.55. 354-359 (1995)
[文献書誌] Matsuhisa, A. et al.: "Inositol monophoshatase activity from the Escherichia coli suhB gene product." J. Bacteriol.177. 200-205 (1995)
[文献書誌] Uehara, Y. et al.: "Placental defect and embryonic lethality in mice lacking hepatocyte growth factor/scatter factor." Nature. 373. 702-705 (1995)
[文献書誌] Takekura, H. et al.: "Abnormal junctions between surface membrane and sarcoplasmic reticulum in skeltal muscle with a mutation targeted to the ryanodine receptor." Proc. Natl. Acad. Sci. USA. 92. 3381-3385 (1995)
[文献書誌] Takeshima, H. et al.: "Ca2+-induced Ca2+release in myocytes from dyspedic mice lacking the type-1 ryanodine receptor." EMBO J.14. 2999-3006 (1995)
[文献書誌] Miyashiro, I. et al.: "Subcellular localization of the APC protein : Immunoelectron microscopic study of the association of the APC protein with catenin." Oncogene. 11. 89-96 (1995)
[文献書誌] Sakaguchi, S. et al: "Accumulation of proteinase K-resistant PrP is restricted by the expression level of normal PrP in mice inoculated with a mouse-adapted strain of Creutzfeldt-Jakob disease agent." J. Virol.69. 7586-7592 (1995)
[文献書誌] Shiba, K. et al.: "Human alanyl-tRNA synthetase : conservation in evolution of catalytic core and microhelix recognition." Biochemistry. 34. 10340-10349 (1995)
[文献書誌] Takei, Y. et al.: "Synapsin I deficiency results in the structural change in the presynaptic terminal in the murine nervous system." J. Cell. Biol.131. 1789-1800 (1995)
[文献書誌] Nakai, S. et al.: "The POU-domain transcription factor Brn-2 is required for the determination of specific neuronal lineages in the hypothalamus of the mouse." Genes Dev.9. 3109-3121 (1995)
[文献書誌] Muto, Y. et al.: "Growth retardation in human cervical dysplasia-derived cell lines by β-carotene through doun-regulation of epidermal growth factor receptor." Am. J. Clin. Nutr.(in press).
[文献書誌] Matsumoto, M. et al.: "Ataxia and epileptic eizure in mice lacking type 1 inositol 1,4,5-trisphophate receptor." Nature. 379. 168-171 (1996)
[文献書誌] Miyatake, S. et al.: "Induction of G1 arrest by down-regulation of cyclin D3 in T cell hybridomas." J. Exp. Med.182. 401-408 (1995)
[文献書誌] Suzuki, T. et al.: "HTLV-1 Tax protein interacts with cyclin-dependent kinase inhibitor p161NK4A and counteracts its inhibitory activity to CDK4." EMBO J.(in press).
[文献書誌] Munirajan, A. K. et al.: "P53 gene mutations in oralb carcinomas from India." Int. J. Cancer. (in press).
[文献書誌] Takada, S. et al.: "Disruption of the function of tumor-suppressor gene p53 by the hepatitis B virus" Res. Clin. Oncol.(in press).
[文献書誌] Yamato, K. et al.: "A human temprature-sensitive p53 mutant, p53val-138 ; Modulation of the cell cycle, viability and expression of p53-responsive genes." Oncogene. 11. 1-6 (1995)
[文献書誌] Hirano, Y. et al.: "A tempertature sensitive mutant of the human p53, vall38, arrest rat cell growth without induced expression of cip1/waf1/sdi1 after temperature shift-down." Oncogene. 10. 1879-1885 (1995)
[文献書誌] Tsutsumi-Ishii, Y.: "Response of heat shock element (HSE) within the human HSP70 promoter to mutated p53 genes." Cell Growth & Differ.6. 1-8 (1995)
[文献書誌] Hirano, Y. et al.: "Roles of p53 mutation in cell line establihsment and identification of the munimum transactivation and transform suppression domains." Eur. J. Cancer. 32B. 129-135 (1995)
[文献書誌] 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.218. 161-164 (1995)
[文献書誌] Sugito, K. et al.: "Interaction between hsp70 and hsp40, eucaryotic homologues of DnaK and DnaJ, in human cells expressing mutant-type p53." FEBS Lett.358. 161-164 (1995)
[文献書誌] Kanda, T. et al.: "Occurrence of the antibody against humanpapillomavirus type 16 (HPV16) virion protein L2 in patients with cervical cancer and dysplasia." Intervirology. 38. 187-191 (1995)
[文献書誌] Nakagawa, S. et al.: "Mutational analysis of human papillomavirus type 16 E6 protein : Transforming function for human cells and degradation of p53 in vitro." Virology. 212. 535-542 (1995)
[文献書誌] Sermusvitayawong, K. et al.: "Heterogeneity of Cellular Proteins that bind to the central conserved region of p53." Medical J. of Osaka Univ.47. 341-350 (1995)
[文献書誌] Shindo, M. et al.: "Detection of human papillomavirus DNA sequences in oral squamous cell carcinomas and their relationship to p53 and PCNA expression." Cancer. 76. 1513-1521 (1995)
[文献書誌] Kitamura, Y. et al.: "Human endogenous retrovirus K10 encodes a functional integrase." J. Virol.(in press).
[文献書誌] Dobashi, Y. et al.: "Constitutive overexpression of cdk2 inhibits neuronal differentiation of rat pheochromocytoma PC12 cells." J. Biol. Chem.270. 23031-23037 (1995)
[文献書誌] Harada, H. et al.: "Activation of a cell cycle regulated histone gene by the oncogenic transcription factor IRF-2." Nature. 377. 362-365 (1995)
[文献書誌] Kimura, T. et al.: "Essential and non-redundunt roles of p48 (ISGF3g) and IRF-1 in both type I and type II interferon responses, as revealed by gene targetting studies." Genes to Cells. (in press). (1996)

1995 年度 実績報告書

癌抑制遺伝子産物の生理機能

研究代表者

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

研究成果

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

[文献書誌] 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.46. 106-112 (1995)

[文献書誌] Hiwasa, T. et al.: "Inhibition of cysteine proteinases and proteosome by Ras, Ran and cyclin." Proteases Involved in Cancer. 183-187 (1995)

[文献書誌] Suzuki-Takashi, I. et al.: "The interaction of E2F with pRB and p107 are regulated via the phosphorylation of pRB and p107 by cyclin-dependent kinase." Oncogene. 10. 1691-1698 (1995)

[文献書誌] Ueno, Y., et al.: "Solid-phase synthesis of peptides containing O-phosphoryl serine and O-phosphoryl threonine using allyl group for phosphate protection." Bioorg. Med. Chem. Let.5. 823-826 (1995)

[文献書誌] Saijo, M. et al.: "Molecular cloning of a human protein that binds to the retinoblastoma protein and chromosomal mapping." Genomics. 27. 551-519 (1995)

[文献書誌] Taya, Y.: "Cell cycle-dependent phosphorylation of the tumor suppressor RB protein." Molecules and Cells. 5. 191-195 (1995)

[文献書誌] Inoue, A. et al.: "70-kDa heat shock cognate protein directly interacts with N-terminal region of the retinoblastoma gene product pRb-identification of a novel region of pRb mediating protein interaction." J. Biol. Chem.270. 22571-22576 (1995)

[文献書誌] Sakai, Y. et al.: "cDNA sequence, chromosomal localization of a novel human protein RBQ-1 which binds to the retinoblastoma gene product." Genomics. 30. 98-101 (1995)

[文献書誌] Higashi, H. et al.: "Differences in substrate specificity between Cdk2-cyclin A and Cdk2-cyclin E in vitro." Biochem. Biophys. Res. Commun.216. 520-525 (1995)

[文献書誌] Higashi, H. et al.: "Cdk2 forms an inactive complex with cyclin D1 since Cdk2 associated with cyclin D1 is not phosphorylated by Cdk7-cyclin H." Eur. J. Biochem.(in press). (1996)

[文献書誌] Kitagawa, M. et al: "The consensus motif for phosphorylation by cyclin D1-Cdk4 is different from that of phosphorylation by cyclin A/E-Cdk2." EMBO J.(in press).

[文献書誌] Suziki, M. et al.: "An E1A-inducible Sp 1 negative regulator, G10BP represses transcription of the rat fibronectin gene." Mol. Cell. Biol.15. 5423-5433 (1995)

[文献書誌] Shimizu, M. et al.: "The G1/S boundary-specific enhancer of the rat cdc2 promoter." Mol. Cell. Biol.15. 2882-2892 (1995)

[文献書誌] Nakajima, T. et al.: "Adenovirus E1A-induced apoptosis elicits a steep decrease in the topoisomerase II a level during the latent phase." Oncogene. 10. 651-662 (1995)

[文献書誌] Nakamra, T. et al.: "Down-regulation of the cyclin A-promoter in differentiating human embryonal carcinoma cells is mediated by depletion of ATF-1 and ATF-2 in the complex at the ATF/CRE site." Exp. Cell. Res.216. 422-430 (1995)

[文献書誌] Kamada, S. et al.: "bcl-2 Deficiency in mice leads to pleiotropic abnormalities : A ccelerated lymphoid cell death in thymus and spleen, polycystic kidney, hair hypopigmentation, and distorted small intestine." Cancer Res.55. 354-359 (1995)

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

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

[文献書誌] Takekura, H. et al.: "Abnormal junctions between surface membrane and sarcoplasmic reticulum in skeltal muscle with a mutation targeted to the ryanodine receptor." Proc. Natl. Acad. Sci. USA. 92. 3381-3385 (1995)

[文献書誌] Takeshima, H. et al.: "Ca2+-induced Ca2+release in myocytes from dyspedic mice lacking the type-1 ryanodine receptor." EMBO J.14. 2999-3006 (1995)

[文献書誌] Miyashiro, I. et al.: "Subcellular localization of the APC protein : Immunoelectron microscopic study of the association of the APC protein with catenin." Oncogene. 11. 89-96 (1995)

[文献書誌] Sakaguchi, S. et al: "Accumulation of proteinase K-resistant PrP is restricted by the expression level of normal PrP in mice inoculated with a mouse-adapted strain of Creutzfeldt-Jakob disease agent." J. Virol.69. 7586-7592 (1995)

[文献書誌] Shiba, K. et al.: "Human alanyl-tRNA synthetase : conservation in evolution of catalytic core and microhelix recognition." Biochemistry. 34. 10340-10349 (1995)

[文献書誌] Takei, Y. et al.: "Synapsin I deficiency results in the structural change in the presynaptic terminal in the murine nervous system." J. Cell. Biol.131. 1789-1800 (1995)

[文献書誌] Nakai, S. et al.: "The POU-domain transcription factor Brn-2 is required for the determination of specific neuronal lineages in the hypothalamus of the mouse." Genes Dev.9. 3109-3121 (1995)

[文献書誌] Muto, Y. et al.: "Growth retardation in human cervical dysplasia-derived cell lines by β-carotene through doun-regulation of epidermal growth factor receptor." Am. J. Clin. Nutr.(in press).

[文献書誌] Matsumoto, M. et al.: "Ataxia and epileptic eizure in mice lacking type 1 inositol 1,4,5-trisphophate receptor." Nature. 379. 168-171 (1996)

[文献書誌] Miyatake, S. et al.: "Induction of G1 arrest by down-regulation of cyclin D3 in T cell hybridomas." J. Exp. Med.182. 401-408 (1995)

[文献書誌] Suzuki, T. et al.: "HTLV-1 Tax protein interacts with cyclin-dependent kinase inhibitor p161NK4A and counteracts its inhibitory activity to CDK4." EMBO J.(in press).

[文献書誌] Munirajan, A. K. et al.: "P53 gene mutations in oralb carcinomas from India." Int. J. Cancer. (in press).

[文献書誌] Takada, S. et al.: "Disruption of the function of tumor-suppressor gene p53 by the hepatitis B virus" Res. Clin. Oncol.(in press).

[文献書誌] Yamato, K. et al.: "A human temprature-sensitive p53 mutant, p53val-138 ; Modulation of the cell cycle, viability and expression of p53-responsive genes." Oncogene. 11. 1-6 (1995)

[文献書誌] Hirano, Y. et al.: "A tempertature sensitive mutant of the human p53, vall38, arrest rat cell growth without induced expression of cip1/waf1/sdi1 after temperature shift-down." Oncogene. 10. 1879-1885 (1995)

[文献書誌] Tsutsumi-Ishii, Y.: "Response of heat shock element (HSE) within the human HSP70 promoter to mutated p53 genes." Cell Growth & Differ.6. 1-8 (1995)

[文献書誌] Hirano, Y. et al.: "Roles of p53 mutation in cell line establihsment and identification of the munimum transactivation and transform suppression domains." Eur. J. Cancer. 32B. 129-135 (1995)

[文献書誌] 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.218. 161-164 (1995)

[文献書誌] Sugito, K. et al.: "Interaction between hsp70 and hsp40, eucaryotic homologues of DnaK and DnaJ, in human cells expressing mutant-type p53." FEBS Lett.358. 161-164 (1995)

[文献書誌] Kanda, T. et al.: "Occurrence of the antibody against humanpapillomavirus type 16 (HPV16) virion protein L2 in patients with cervical cancer and dysplasia." Intervirology. 38. 187-191 (1995)

[文献書誌] Nakagawa, S. et al.: "Mutational analysis of human papillomavirus type 16 E6 protein : Transforming function for human cells and degradation of p53 in vitro." Virology. 212. 535-542 (1995)

[文献書誌] Sermusvitayawong, K. et al.: "Heterogeneity of Cellular Proteins that bind to the central conserved region of p53." Medical J. of Osaka Univ.47. 341-350 (1995)

[文献書誌] Shindo, M. et al.: "Detection of human papillomavirus DNA sequences in oral squamous cell carcinomas and their relationship to p53 and PCNA expression." Cancer. 76. 1513-1521 (1995)

[文献書誌] Kitamura, Y. et al.: "Human endogenous retrovirus K10 encodes a functional integrase." J. Virol.(in press).

[文献書誌] Dobashi, Y. et al.: "Constitutive overexpression of cdk2 inhibits neuronal differentiation of rat pheochromocytoma PC12 cells." J. Biol. Chem.270. 23031-23037 (1995)

[文献書誌] Harada, H. et al.: "Activation of a cell cycle regulated histone gene by the oncogenic transcription factor IRF-2." Nature. 377. 362-365 (1995)

[文献書誌] Kimura, T. et al.: "Essential and non-redundunt roles of p48 (ISGF3g) and IRF-1 in both type I and type II interferon responses, as revealed by gene targetting studies." Genes to Cells. (in press). (1996)

1995 年度実績報告書

田矢洋一国立がんセンター, 研究所生物学部, 室長 (60133641)