1999 Fiscal Year Annual Research Report

ヒト紫外線感受性を一本の毛髪を用いて評価するシステムの開発

Research Project

Project/Area Number	11558067
Research Category	Grant-in-Aid for Scientific Research (B)
Research Institution	Tohoku University
Principal Investigator	山本和生東北大学, 大学院・理学研究科, 教授 (20093536)
Co-Investigator(Kenkyū-buntansha)	竹内栄岡山大学, 理学部, 助手 (20226989) 井出宏之東北大学, 大学院・理学研究科, 教授 (70022704) 山本博章東北大学, 大学院・理学研究科, 助教授 (40174809) 鈴木格ポーラ化成工業株式会社, 皮膚科学研究部, 研究員
Keywords	DNA修復能 / 色素生産能 / ピリミジン2量体 / 6-4付加体 / チロシナーゼ / メラニン産生 / αMSH / 固相化
Research Abstract	本研究計画は、ヒト個々人の紫外線感受性を簡便に評価するシステムの開発を目標にしている。今年度はこの感受性に関わるDNA修復能やメラニン色素産生能に関わるタンパク質に対する抗体を調製することが第一義的な目的であった。紫外線による代表的損傷ピリミジン2量体と6-4付加体をそれぞれ修復する光回復酵素の遺伝子をカンガルーとシロイヌナズナからクローニングし、大腸菌で発現させ、精製した酵素標品を用いて抗体を作成した。また、ピリミジン2量体と6-4付加体に対する抗体を金沢大学二階堂教授の研究室より分与を受け、損傷定量のシステムを完成した。一方メラニン産生の鍵酵素チロシナーゼ、またそのパラログで、TRP1(tyrosinase-relatedprotein1,チロシナーゼ関連配列)に対する抗体は入手した。TRP-2についてはタンパク質合成の後、ポリクローナル抗体を作製すべく現在cDNAをクローニングしている。断片がすでに調製できたので全長のクローニングを行っている。またヒト皮膚の色素細胞に作用してメラニン産生を亢進するホルモンとしてαMSHが知られているが、このホルモンの作用機序を支えている因子として、p(pink-eyed dilution)遺伝子産物の関与に気付いたので、当該タンパク質についても抗体を作製すべくまずcDNA断片のクローニングを行った。現在この全長の調製も行っている。次に抗体を固相化し、これを用いて上記遺伝子産物をアッセイするシステムを開発することを試みている。固相化そのものはいろいろ方法が利用できるが、将来的には毛根を圧着させてスタンプする予定であるので、膜強度の問題を克服しなければならない状況である。いずれにしてもいくつかの候補遺伝子が揃いつつあるので、まず簡便なmRNAの半定量的アッセイキットの開発も検討しているところである。

Research Products
(26 results)

All Other

All Publications (26 results)

[Publications] T.Nunoshiba: "Role of iron and superoxide for generation of hydroxyl radical, oxidative DNA lesions and mutagenesis in Esherichia coli"J. Biol. Chem.. 274. 34832-34837 (1999)
[Publications] N.Uematsu: "Asymmetric crossover for spontaneous formation of large deletions at the tonB-trp region of Escherichia coli K-12 cells"Mol. Gen. Genet. 261. 523-529 (1999)
[Publications] T.Funayama: "Deinococcus radiodurans recN gene: A molecular analysis of mitomycin-C sensitive strain and gene"Mutation Res.. 435. 151-16* (1999)
[Publications] H.Iyehara-Ogawa: "Cobaltous chloride-induced mutagenesis in the supF tRNA gene of Escherichia coli"Mutagenesis. 14. 249-253 (1999)
[Publications] T.Nunoshiba: "Role of glutathione on acrolein-induced cytotoxicity and mutagenicity in Escherichia coli"Mutation Res.. 442. 1-8 (1999)
[Publications] Y.Agemizu: "DNA sequence analysis of spontanous tonB deletion mutations in a polA1 strain of Escherichia coli K12"Biochem. Biophys. Res. Commun. 261. 584-589 (1999)
[Publications] M.Kikuchi: "Genomic organization of the extraordinary radioresistant bacterium Deinococcus radiodurans: physical map and evidence for multiple replicons"FEMS Microbiol. Letters. 174. 151-157 (1999)
[Publications] T.Ono: "Molecular nature of mutation induced by high dose of X-ray in spleen, liver and brain of the lacZ-transgenic mouse"Environmental and Molecular Mutagenesis. 34. 97-105 (1999)
[Publications] I.Narumi: "Molecular analysis of Deinococcus radiodurans recA locus and identification of a mutation site in a DNA-repair deficient mutant rec30"Mutation Res.. 435. 233-243 (1999)
[Publications] T.Najrana: "Spontaneous and osmium tetroxide induced mutagenesis in Escherichia coli strain deficient both endonuclease III and endonuclease VIII"Mutagenesis. (印刷中). (1999)
[Publications] T.Ono: "Age-associated increase of spontaneous mutation frequency and molecular nature of mutation in newborn and old lacZ-transgenic mouse"Mutation Res.. (印刷中). (1999)
[Publications] S.Nakamura: "Radiation-induced mutation in spleen and brain of lacZ transgenic mouse"Int. J. Radiat. Biol.. (印刷中). (1999)
[Publications] Y.Manabe: "Immunocytochemical and immuno-electron-microscopical study of somatotrophs in ICR and non-obese diabetic (NOD) mice"Cells Tissues Organs. 166. 31-39 (2000)
[Publications] I.Suzuki: "Participation od the melanocortin-1 receptor in the UV control of pigmentaiton"J. Invest. Dermatol. Symp. Proc.. 4. 29-34 (1999)
[Publications] H.Yokoyama: "Mesenchyme with fgf-10 Expression Is Responsible for Regenerative Capacity in Xenopus Limb Buds"Dev Biol. 219. 18-29 (2000)
[Publications] H.Yajima: "Role of N-cadherin in the sorting-out of mesenchymal cells and in the positional identity along the proximodistal axis of the chick limb bud"Dev Dyn. 216. 274-284 (1999)
[Publications] S.Yonei-Tamura: "FGF7 and FGF10 directly induce the apical ectodermal ridge in chick embryos"Dev Biol. 211. 133-143 (1999)
[Publications] I.Yajima: "An L1 element intronic insertion in the black-eyed white (Mitfmi-bw) gene: the loss of a single Mitf isoform responsible for the pigmentary defect and inner ear deafness"Hum. Mol. Genet.. 8. 1431-1441 (1999)
[Publications] S.Takeuchi: "Widespread expression of Agouti-related proteiin (AGRP) in the chicken: A possible involvement of AGRP in regulating peripheral melanocortin systems in the chicken"Biochim. Biophys. Acta.. (印刷中).
[Publications] S.Takeuchi: "A possible involvement of melanocortin 3-receptor in the regulation of adrenal gland function in the chicken"Biochim. Biophys. Acta. 1448. 512-518 (1999)
[Publications] S.Takeuchi: "Molecular cloning and characterization of the chicken pro-opiomelanocortin (POMC)"Biochim. Biophys. Acta. 1450. 452-459 (1999)
[Publications] S.Takeuchi: "A possible mechanism for feedback regulation of the mouse tyrosinase gene by its 3' non-coding RNA fragments"Pigment Cell Res. (印刷中).
[Publications] T.Kakeya: "Epidermal growth factor, insulin and estrogen stimulate development of prolactinsecreting cells in cultures of GH3 cells"Cell Tissue Res. (印刷中).
[Publications] M.Shiraga: "Epidermal growth factor stimulates proliferation of mouse uterine epithelial cells in primary culture"Zool.Sci.. (印刷中).
[Publications] H.Yuasa: "The structural organization of the ascidian, Halocynthia roretzi calmodulin genes: The vicissitude of introns during the evolution of calmodulin genes"Gene. 229. 163-169 (1999)
[Publications] S.Sato: "Structure and developmental expression of the ascidian TRP gene: insights into the evolution of pigment cell-specific gene expression"Dev. Dyn.. 25. 225-237 (1999)

1999 Fiscal Year Annual Research Report

ヒト紫外線感受性を一本の毛髪を用いて評価するシステムの開発

Principal Investigator

山本 和生 東北大学, 大学院・理学研究科, 教授 (20093536)

Research Products

[Publications] T.Nunoshiba: "Role of iron and superoxide for generation of hydroxyl radical, oxidative DNA lesions and mutagenesis in Esherichia coli"J. Biol. Chem.. 274. 34832-34837 (1999)

[Publications] N.Uematsu: "Asymmetric crossover for spontaneous formation of large deletions at the tonB-trp region of Escherichia coli K-12 cells"Mol. Gen. Genet. 261. 523-529 (1999)

[Publications] T.Funayama: "Deinococcus radiodurans recN gene: A molecular analysis of mitomycin-C sensitive strain and gene"Mutation Res.. 435. 151-16* (1999)

[Publications] H.Iyehara-Ogawa: "Cobaltous chloride-induced mutagenesis in the supF tRNA gene of Escherichia coli"Mutagenesis. 14. 249-253 (1999)

[Publications] T.Nunoshiba: "Role of glutathione on acrolein-induced cytotoxicity and mutagenicity in Escherichia coli"Mutation Res.. 442. 1-8 (1999)

[Publications] Y.Agemizu: "DNA sequence analysis of spontanous tonB deletion mutations in a polA1 strain of Escherichia coli K12"Biochem. Biophys. Res. Commun. 261. 584-589 (1999)

[Publications] M.Kikuchi: "Genomic organization of the extraordinary radioresistant bacterium Deinococcus radiodurans: physical map and evidence for multiple replicons"FEMS Microbiol. Letters. 174. 151-157 (1999)

[Publications] T.Ono: "Molecular nature of mutation induced by high dose of X-ray in spleen, liver and brain of the lacZ-transgenic mouse"Environmental and Molecular Mutagenesis. 34. 97-105 (1999)

[Publications] I.Narumi: "Molecular analysis of Deinococcus radiodurans recA locus and identification of a mutation site in a DNA-repair deficient mutant rec30"Mutation Res.. 435. 233-243 (1999)

[Publications] T.Najrana: "Spontaneous and osmium tetroxide induced mutagenesis in Escherichia coli strain deficient both endonuclease III and endonuclease VIII"Mutagenesis. (印刷中). (1999)

[Publications] T.Ono: "Age-associated increase of spontaneous mutation frequency and molecular nature of mutation in newborn and old lacZ-transgenic mouse"Mutation Res.. (印刷中). (1999)

[Publications] S.Nakamura: "Radiation-induced mutation in spleen and brain of lacZ transgenic mouse"Int. J. Radiat. Biol.. (印刷中). (1999)

[Publications] Y.Manabe: "Immunocytochemical and immuno-electron-microscopical study of somatotrophs in ICR and non-obese diabetic (NOD) mice"Cells Tissues Organs. 166. 31-39 (2000)

[Publications] I.Suzuki: "Participation od the melanocortin-1 receptor in the UV control of pigmentaiton"J. Invest. Dermatol. Symp. Proc.. 4. 29-34 (1999)

[Publications] H.Yokoyama: "Mesenchyme with fgf-10 Expression Is Responsible for Regenerative Capacity in Xenopus Limb Buds"Dev Biol. 219. 18-29 (2000)

[Publications] H.Yajima: "Role of N-cadherin in the sorting-out of mesenchymal cells and in the positional identity along the proximodistal axis of the chick limb bud"Dev Dyn. 216. 274-284 (1999)

[Publications] S.Yonei-Tamura: "FGF7 and FGF10 directly induce the apical ectodermal ridge in chick embryos"Dev Biol. 211. 133-143 (1999)

[Publications] I.Yajima: "An L1 element intronic insertion in the black-eyed white (Mitfmi-bw) gene: the loss of a single Mitf isoform responsible for the pigmentary defect and inner ear deafness"Hum. Mol. Genet.. 8. 1431-1441 (1999)

[Publications] S.Takeuchi: "Widespread expression of Agouti-related proteiin (AGRP) in the chicken: A possible involvement of AGRP in regulating peripheral melanocortin systems in the chicken"Biochim. Biophys. Acta.. (印刷中).

[Publications] S.Takeuchi: "A possible involvement of melanocortin 3-receptor in the regulation of adrenal gland function in the chicken"Biochim. Biophys. Acta. 1448. 512-518 (1999)

[Publications] S.Takeuchi: "Molecular cloning and characterization of the chicken pro-opiomelanocortin (POMC)"Biochim. Biophys. Acta. 1450. 452-459 (1999)

[Publications] S.Takeuchi: "A possible mechanism for feedback regulation of the mouse tyrosinase gene by its 3' non-coding RNA fragments"Pigment Cell Res. (印刷中).

[Publications] T.Kakeya: "Epidermal growth factor, insulin and estrogen stimulate development of prolactinsecreting cells in cultures of GH3 cells"Cell Tissue Res. (印刷中).

[Publications] M.Shiraga: "Epidermal growth factor stimulates proliferation of mouse uterine epithelial cells in primary culture"Zool.Sci.. (印刷中).

[Publications] H.Yuasa: "The structural organization of the ascidian, Halocynthia roretzi calmodulin genes: The vicissitude of introns during the evolution of calmodulin genes"Gene. 229. 163-169 (1999)

[Publications] S.Sato: "Structure and developmental expression of the ascidian TRP gene: insights into the evolution of pigment cell-specific gene expression"Dev. Dyn.. 25. 225-237 (1999)

山本和生東北大学, 大学院・理学研究科, 教授 (20093536)