1999 Fiscal Year Annual Research Report

突然変異の分子病態

Research Project

Project/Area Number	08280104
Research Institution	Nara Institute of Science and Technology
Principal Investigator	真木寿治奈良先端科学技術大学院大学, バイオサイエンス研究科, 教授 (20199649)
Co-Investigator(Kenkyū-buntansha)	続輝久九州大学, 医学部, 教授 (40155429) 葛西宏産業医科大学, 産業生態科学研究所, 教授 (40152615) 杉野明雄大阪大学, 微生物病研究所, 教授 (90231737) 能美健彦国立医薬品食品衛生研究所, 変異遺伝部, 室長(研究職) (30150890) 中村健司東京大学, 医科学研究所, 助手 (90253533)
Keywords	DNA複製 / 複製エラー / ミスマッチ修復 / 酵素ラジカル / DNA修復 / フレームシフト / 繰り返し配列 / ノックアウトマウス
Research Abstract	本研究計画での主要な問題点は自然突然変異の発生原因や変異を抑制する機構の解明と生体でのそれらの制御や生物学的あるいは医学的影響を明らかにすることである。本年度は以下の2つの項目について研究を実施し、新たな知見を得た。 1.自然突然変異の分子機構:DNAポリメラーゼによる複製エラーを解析するための新しい試験管内複製系を構築し、lagging鎖DNA合成での複製エラーの特徴を検討した。これまでの複製フォークでのDNA合成に比較して、lagging鎖合成では10倍程度正確な複製がなされることが判明した。また、校正機能が欠損した場合の複製エラーの特徴についても明らかにし、顕著な鋳型配列依存性が見出された。大腸菌およびヒト細胞において、DNA損傷でのバイパスDNA合成に関与する新規のDNAポリメラーゼを発見し、それらの生化学的特徴付けを行った。酵素ラジカルによる異変原性ヌクレオチドおよびDNA損傷について、試験管内DNA合成系での異変誘発能を詳細に検討した。 2.高等生物における突然変異の特異性:酵素ラジカルによる損傷に起因する突然変異を解析するためにMTH1遺伝子、MSH2遺伝子、および両遺伝子欠損のノックアウトマウスに異変解析用の標的遺伝子を導入した。このマウスでの突然変異率と異変スペクトラムの検討を開始した。染色体上のマイクロサテライトDNAの繰り返し数の変化を引き起こしている変異の発生について、癌関連遺伝子およびそれ以上の遺伝子での発生頻度を検討し、ヒトでのこれらの異変の意義について検討した。

Research Products
(20 results)

All Other

All Publications (20 results)

[Publications] S.Fujii: "DNA Replication Errors Produced by the Replicative Apparatus of Esherichia coil"J.Mol.Biol.. 289. 835-850 (1999)
[Publications] M.Seki: "Strand Asymmetry of+1 Frameshift Mutagenesis at Repetitive DNA Sequences by DNA Polymerase III Holoenzyme of Escherichia coil"J.Biol.Chem.. 274. 33313-33319 (1999)
[Publications] H.Masumoto: "Dpb11 controls the association between DNA polymerase alpha and epsilon, and the ARS region of budding yeast"H.Mol.Cell.Biol.. (in press). (2000)
[Publications] L.H.Johnston: "First the CDKs,now the DDKs"Trends Cell Biol. 9. 249-252 (1999)
[Publications] K.Fujikawa: "The oxidized forms of dATP are substrates for the human MutT homologue,the hMTH1 protein"J.Biol.Chem.. 274. 18201-18205 (1999)
[Publications] M.Nomoto: "Analysis of 8-hydroxyguanine in rat kidney genomic DNA after administration of renal carcinogen,ferric nitriloacetate"Carcinogenesis. 20. 837-841 (1999)
[Publications] K.Nishii: "Abnormalities of developmental cell death in Dad1-deficient mice"Genes to Cells. 4. 243-252 (1999)
[Publications] Y.Yanagawa: "Enrichment and efficient screening of ES cells containing a targeted mutation:the use of DT-A gene with the polyadenylation single as a negative selection marker"Transgenic Res.. 8. 215-221 (1999)
[Publications] T.Sato: "Transgenic mice harboring a full-length human mutant DRPLA gene exhibit age-dependent intergenerational and somatic instabilities of CAG repeats comparable with those in DRPLA patients"Human Molecular Genetics. 8(1). 99-106 (1999)
[Publications] A.Naito: "Severe osteopetrosis,defective interleukin-1 signalling and lymph node organogenesis in TRAF6-deficient mice"Genes to Cells. 4(6). 353-362 (1999)
[Publications] J.Wagner: "The dinB gene encodes a novel Escherichia coil DNA polymerase(DNA pol IV) involved in mutagenesis"Mol.Cell. 4. 281-286 (1999)
[Publications] K.Masumura: "Spectra of gpt mutations in ethylnitrosourea-treated and untreated transgenic mice"Env.Mol.Mutagen.. 34. 1-8 (1999)
[Publications] K.Hayashi: "Recent enhancements in SSCP"Genetic Analysis:Biomolecular Engineering. 14. 193-196 (1999)
[Publications] T.Horiuchi: "Association of Fas/APO-1 gene polymorphism with systemic lupus erythematosus in Japanese"Reumatology. 38. 516-520 (1999)
[Publications] N.Azuma: "Missense mutation in the alternative splice region of the PAX6 gene in eye anomalies"Am.J.Hum.Genet.. 65. 656-663 (1999)
[Publications] K.Negishi: "Various phenotypic expresion of familial aniridia with a PAX6 mutation"Brit.J.Ophthalmol. 83. 991-992 (1999)
[Publications] T.Ogi: "Mutation enhancement by DINB1,a mammalian homologue of the Escherichia coil"Genes to Cells. 4(11). 607-618 (1999)
[Publications] R.Antonio: "Identification of additional genes belonging to the LexA regulon in Escherichia coil mutagenesis protein DinB"mokecular Microbiology. 120(in press). (2000)
[Publications] D.Matsuzoe: "p53 mutations predict non-small cell lung carcinoma response to radiotherapy"Cancer Letter. 135. 189-194 (1999)
[Publications] D.Matsuzoe: "Discrimination of double primary lung cancer from intrapulmonary metastasis by p53 gene mutation"Br.J.Cancer. 79. 1549-1552 (1999)

1999 Fiscal Year Annual Research Report

突然変異の分子病態

Principal Investigator

真木 寿治 奈良先端科学技術大学院大学, バイオサイエンス研究科, 教授 (20199649)

Research Products

[Publications] S.Fujii: "DNA Replication Errors Produced by the Replicative Apparatus of Esherichia coil"J.Mol.Biol.. 289. 835-850 (1999)

[Publications] M.Seki: "Strand Asymmetry of+1 Frameshift Mutagenesis at Repetitive DNA Sequences by DNA Polymerase III Holoenzyme of Escherichia coil"J.Biol.Chem.. 274. 33313-33319 (1999)

[Publications] H.Masumoto: "Dpb11 controls the association between DNA polymerase alpha and epsilon, and the ARS region of budding yeast"H.Mol.Cell.Biol.. (in press). (2000)

[Publications] L.H.Johnston: "First the CDKs,now the DDKs"Trends Cell Biol. 9. 249-252 (1999)

[Publications] K.Fujikawa: "The oxidized forms of dATP are substrates for the human MutT homologue,the hMTH1 protein"J.Biol.Chem.. 274. 18201-18205 (1999)

[Publications] M.Nomoto: "Analysis of 8-hydroxyguanine in rat kidney genomic DNA after administration of renal carcinogen,ferric nitriloacetate"Carcinogenesis. 20. 837-841 (1999)

[Publications] K.Nishii: "Abnormalities of developmental cell death in Dad1-deficient mice"Genes to Cells. 4. 243-252 (1999)

[Publications] Y.Yanagawa: "Enrichment and efficient screening of ES cells containing a targeted mutation:the use of DT-A gene with the polyadenylation single as a negative selection marker"Transgenic Res.. 8. 215-221 (1999)

[Publications] T.Sato: "Transgenic mice harboring a full-length human mutant DRPLA gene exhibit age-dependent intergenerational and somatic instabilities of CAG repeats comparable with those in DRPLA patients"Human Molecular Genetics. 8(1). 99-106 (1999)

[Publications] A.Naito: "Severe osteopetrosis,defective interleukin-1 signalling and lymph node organogenesis in TRAF6-deficient mice"Genes to Cells. 4(6). 353-362 (1999)

[Publications] J.Wagner: "The dinB gene encodes a novel Escherichia coil DNA polymerase(DNA pol IV) involved in mutagenesis"Mol.Cell. 4. 281-286 (1999)

[Publications] K.Masumura: "Spectra of gpt mutations in ethylnitrosourea-treated and untreated transgenic mice"Env.Mol.Mutagen.. 34. 1-8 (1999)

[Publications] K.Hayashi: "Recent enhancements in SSCP"Genetic Analysis:Biomolecular Engineering. 14. 193-196 (1999)

[Publications] T.Horiuchi: "Association of Fas/APO-1 gene polymorphism with systemic lupus erythematosus in Japanese"Reumatology. 38. 516-520 (1999)

[Publications] N.Azuma: "Missense mutation in the alternative splice region of the PAX6 gene in eye anomalies"Am.J.Hum.Genet.. 65. 656-663 (1999)

[Publications] K.Negishi: "Various phenotypic expresion of familial aniridia with a PAX6 mutation"Brit.J.Ophthalmol. 83. 991-992 (1999)

[Publications] T.Ogi: "Mutation enhancement by DINB1,a mammalian homologue of the Escherichia coil"Genes to Cells. 4(11). 607-618 (1999)

[Publications] R.Antonio: "Identification of additional genes belonging to the LexA regulon in Escherichia coil mutagenesis protein DinB"mokecular Microbiology. 120(in press). (2000)

[Publications] D.Matsuzoe: "p53 mutations predict non-small cell lung carcinoma response to radiotherapy"Cancer Letter. 135. 189-194 (1999)

[Publications] D.Matsuzoe: "Discrimination of double primary lung cancer from intrapulmonary metastasis by p53 gene mutation"Br.J.Cancer. 79. 1549-1552 (1999)

真木寿治奈良先端科学技術大学院大学, バイオサイエンス研究科, 教授 (20199649)