1990 Fiscal Year Annual Research Report

蛋白質/DNA複合体の構造

Research Project

Project/Area Number	02263102
Research Institution	Osaka University
Principal Investigator	箱嶋敏雄大阪大学, 薬学部, 助手 (00164773)
Co-Investigator(Kenkyū-buntansha)	広瀬進国立遺伝学研究所, 遺伝情報研究センター, 助教授 (90022730) 月原冨武鳥取大学, 工学部, 助教授 (00032277) 田中勲北海道大学, 理学部, 助教授 (70093052) 坂部知平高エネルギー物理学研究所, 放射光実験施設, 教授 (30022601) 小川智子大阪大学, 理学部, 講師 (80028208)
Keywords	蛋白質構造 / DNA構造 / 分子認識 / 三次元構造 / 高次構造 / 転写制御 / 塩基配列特異性 / DNA結合蛋白質
Research Abstract	塩基配列に特異的なDNA結合蛋白質として、分裂酵母のAP1様転写制御因子papl+、ヒトのインタ-フェロン関連遺伝子の転写制御因子IRF2、大腸菌の浸透圧レギュロンの転写因子OmpR、大腸菌の複製終結点結合因子Tarについて、また、DNA塩基配列には非特異的な結合蛋白質として大腸菌のクロモゾ-マル蛋白質HーNS、大腸菌の単鎖DNA結合蛋白質SSB、RNAと結合する蛋白質として大腸菌のRAS/EFーTu様蛋白質EraはDNA結合ドメインのサブクロ-ニングや大量精製を終了し結晶化の段階に至った。特に、SSBについては、結晶化に成功し、シンクロトロン放射X線を用いて分解能4A^^°までの回折デ-タを収集した。また、HーNSやEraについては微結晶や球晶の観察で結晶成長の可能性が確認された。酵母リボゾ-ムの大量精製を行い、80S粒子が安定に存在できる条件を見いだした。X線構造解析としては、好熱菌HUの高分解能(2A^^°)での構造精密化を行った。大腸菌HUと合成DNAの複合体結晶は分解能3.7A^^°でR値が29%まで構造を精密化した。イメ-ジングプレ-ト(1P)からの強度読み取り機の導入により、IPを装着した巨大ワイセンベルグ型X線カメラを用いたシンクロトロン放射X線による迅速X線回折強度デ-タ収集の効率化を計った。カイコ絹糸腺抽出液中に検出したDNA超らせん形成する因子(酸性蛋白質、50KDa)をクロ-ニングしつつあり、この因子の活性へのデキストラン硫酸などポリアニオンの効果を調べた。超らせんによる転写の律速段階であるTFIID/プロモ-タ複合体形成の促進効果を種々のプロモ-タを用いて検討し、その普遍性を確認した。大腸菌RecAのキメラ分子を数多く作成し、大きくは3つドメインをもち、中央のドメインは4つの領域に分かれることが明らかになった。

Research Products

(29 results)

All Other

All Publications (29 results)

[Publications] T.Hakoshima: "Crystallization and preliminary Xーray investigation of nonspecific complexes of a mutant ribonuclease T_1 (Y45W) with 2'AMP and 2'UMP" <J.Mol.Biol.>___ー. 216. 497-499 (1990)
[Publications] T.Hakoshima: "Crystallographic characterization of the mutant and wildーtype ribonuclease T_1 complexed with sevaral ribonucleotides" <J.Biochem (Tokyo)>___ー. 108. 695-698 (1990)
[Publications] S.Nishikawa: "A thermostable mutant of ribonuclease T_1 having three disulfide bond" <Protein Engineering>___ー. 3. 443-448 (1990)
[Publications] 箱嶋敏雄: "遺伝子発現調節に関与する蛋白質の構造研究" 日本結晶学会誌. 32. 311-314 (1990)
[Publications] M.Kojima: "Interaction of RecA protein with pBR322 DNA modified by Nーhydroxyー2ーacetlyaminofluorene and 4ーhydroxyaminoquinoline 1ーoxide" <Nucle.Acids Res.>___ー. 18. 2707-2714 (1990)
[Publications] A.Higashitani: "ATPーIndependent strand transfer protein from murine spermatocytes,spermatids,and spermatozoa" <Experimental Cell Res.>___ー. 186. 317-323 (1990)
[Publications] H.Ogawa: "Regulation in repressor interaction by RecA protein" <Adv.Biophys.>___ー. 26. 33-48 (1990)
[Publications] 小川智子: "大腸菌SOS応答反応の分子機構" 蛋白質核酸酵素. 35. 893-904 (1990)
[Publications] N.Sakabe: "Protein crystallography using imaging plates and synchrotoron radiation in the photon factory" <Acta Crystallogr.>___ー. A46. C8 (1990)
[Publications] N.Sakabe: "Crystal structure analysis of chicken gizzard Gーactin DNase I complex at 2.4A^^° resolution" <Acta Crystallogr.>___ー. A46. C90 (1990)
[Publications] N.Watanabe: "Crystal structure analysis of ωーamino acid:pyruvata aminotransferase and observation of change in diffraction by time resolution Laue method" <Acta Crystallogr.>___ー. A46. C19 (1990)
[Publications] I.Tanaka: "An automatic diffraction data collection system with an imaging plate" <J.Appl.Crystallogr.>___ー. 23. 334-339 (1990)
[Publications] 田中勲: "DNA結合蛋白質HU立体構造" 生物物理. 29. 6-11 (1990)
[Publications] T.Tsukihara: "Symmetry and subunit arrengement of Tobacco necrosis virus (TNV)" <Acta Crystallogr.>___ー. B46. 855-860 (1990)
[Publications] T.Ohta: "Purification of DNA supercoiling factor from the posterior silk gland of <Bombyx mori>___ー" <Proc.Natl.Acad.Sci.USA>___ー. 87. 5307-5311 (1990)
[Publications] T.Tamura: "Core promoter of the mouse myelin basic protein gene governs brainーspecific transcription <in vitro>___ー" <EMBO J.>___ー. 9. 3101-3108 (1990)
[Publications] S.Hirose: "DNA supercoling and eukaryotic transcriptionーCause and Effect" <Cell structure and Function>___ー. 15. 133-135 (1990)
[Publications] K.Katayanagi: "Threeーdimensional structure of ribonuclease H from <E.coli>___ー" <Nature>___ー. 347. 306-309 (1990)
[Publications] H.Ueda: "Identification and purification of a <Bombyx mori>___ー homologue of FTZーF1" <Nucle.Acids Res.>___ー. 18. 7229-7234 (1990)
[Publications] K.Wilson: "The termostability of DNAーbinding protein HU from <Bacilli>___ー" <Protein Eng.>___ー. 4. 11-22 (1990)
[Publications] K.Okada: "Negative supercoling of DNA by eukaryotic DNA topoisomerase II and dextran sulfate" <J.Biochem.>___ー. 109. 365-369 (1991)
[Publications] K.Terada: "Demonstration of extrinsic DNA from immune complexes in plasma of a patient with systemic lupus erythematosus" <Biochem.Biophys.REs.Commun>___ー.174. 323-330 (1991)
[Publications] H.Ohishi: "Interaction between the leftーhanded ZーDNA and polyamine:The crystal structure of the d(CG)_3 and Nー(2ーaminoethyl)ー1,4ーdiaminobutane complex" <FEBS Lett>___ー.,. (1991)
[Publications] M.Mizutani: "Negative supercoling of DNA facilitates an interaction between transcription factor IID and the fibroin gene promotor" <Proc.Natl.Acad.Sci>___ー.(USA). 88. (1991)
[Publications] T.Horii: "Mapping of functional domains by truncated by RecA proteins" <J.Mol.Biol>___ー.(1991)
[Publications] Tateishi: "Cーterminal truncated <E.coli>___ー RecA protein,RecA_<5327>,has enhanced DNA binding affinities to singleーand doubleーstranded DNAs:Reguratory role of Cーterminal region in DNA binding" <J.Mol.Biol>___ー.(1991)
[Publications] 箱嶋敏雄、池原森男 (分担執筆): "分子生物の進歩第3巻"蛋白質の機能構造" リボヌクレア-ゼの機能構造" 丸善(日本分子生物学会編), 31 (1990)
[Publications] 箱嶋敏雄、冨田研一 (分担執筆): "<新生化学実験講座第2巻"核酸II構造と性質">___ー X線回折" 東京化学同人(日本生化学会編), (1991)
[Publications] 小川智子 (分担執筆): "<新生化学実験講座核酸の化学I>___ー密度勾配法" 東京化学同人(生化学会編), (1991)

1990 Fiscal Year Annual Research Report

蛋白質/DNA複合体の構造

Principal Investigator

箱嶋敏雄大阪大学, 薬学部, 助手 (00164773)

Research Products

[Publications] T.Hakoshima: "Crystallization and preliminary Xーray investigation of nonspecific complexes of a mutant ribonuclease T_1 (Y45W) with 2'AMP and 2'UMP" <J.Mol.Biol.>___ー. 216. 497-499 (1990)

[Publications] T.Hakoshima: "Crystallographic characterization of the mutant and wildーtype ribonuclease T_1 complexed with sevaral ribonucleotides" <J.Biochem (Tokyo)>___ー. 108. 695-698 (1990)

[Publications] S.Nishikawa: "A thermostable mutant of ribonuclease T_1 having three disulfide bond" <Protein Engineering>___ー. 3. 443-448 (1990)

[Publications] 箱嶋敏雄: "遺伝子発現調節に関与する蛋白質の構造研究" 日本結晶学会誌. 32. 311-314 (1990)

[Publications] M.Kojima: "Interaction of RecA protein with pBR322 DNA modified by Nーhydroxyー2ーacetlyaminofluorene and 4ーhydroxyaminoquinoline 1ーoxide" <Nucle.Acids Res.>___ー. 18. 2707-2714 (1990)

[Publications] A.Higashitani: "ATPーIndependent strand transfer protein from murine spermatocytes,spermatids,and spermatozoa" <Experimental Cell Res.>___ー. 186. 317-323 (1990)

[Publications] H.Ogawa: "Regulation in repressor interaction by RecA protein" <Adv.Biophys.>___ー. 26. 33-48 (1990)

[Publications] 小川智子: "大腸菌SOS応答反応の分子機構" 蛋白質核酸酵素. 35. 893-904 (1990)

[Publications] N.Sakabe: "Protein crystallography using imaging plates and synchrotoron radiation in the photon factory" <Acta Crystallogr.>___ー. A46. C8 (1990)

[Publications] N.Sakabe: "Crystal structure analysis of chicken gizzard Gーactin DNase I complex at 2.4A^^° resolution" <Acta Crystallogr.>___ー. A46. C90 (1990)

[Publications] N.Watanabe: "Crystal structure analysis of ωーamino acid:pyruvata aminotransferase and observation of change in diffraction by time resolution Laue method" <Acta Crystallogr.>___ー. A46. C19 (1990)

[Publications] I.Tanaka: "An automatic diffraction data collection system with an imaging plate" <J.Appl.Crystallogr.>___ー. 23. 334-339 (1990)

[Publications] 田中勲: "DNA結合蛋白質HU立体構造" 生物物理. 29. 6-11 (1990)

[Publications] T.Tsukihara: "Symmetry and subunit arrengement of Tobacco necrosis virus (TNV)" <Acta Crystallogr.>___ー. B46. 855-860 (1990)

[Publications] T.Ohta: "Purification of DNA supercoiling factor from the posterior silk gland of <Bombyx mori>_ー" <Proc.Natl.Acad.Sci.USA>_ー. 87. 5307-5311 (1990)

[Publications] T.Tamura: "Core promoter of the mouse myelin basic protein gene governs brainーspecific transcription <in vitro>_ー" <EMBO J.>_ー. 9. 3101-3108 (1990)

[Publications] S.Hirose: "DNA supercoling and eukaryotic transcriptionーCause and Effect" <Cell structure and Function>___ー. 15. 133-135 (1990)

[Publications] K.Katayanagi: "Threeーdimensional structure of ribonuclease H from <E.coli>_ー" <Nature>_ー. 347. 306-309 (1990)

[Publications] H.Ueda: "Identification and purification of a <Bombyx mori>_ー homologue of FTZーF1" <Nucle.Acids Res.>_ー. 18. 7229-7234 (1990)

[Publications] K.Wilson: "The termostability of DNAーbinding protein HU from <Bacilli>_ー" <Protein Eng.>_ー. 4. 11-22 (1990)

[Publications] K.Okada: "Negative supercoling of DNA by eukaryotic DNA topoisomerase II and dextran sulfate" <J.Biochem.>___ー. 109. 365-369 (1991)

[Publications] K.Terada: "Demonstration of extrinsic DNA from immune complexes in plasma of a patient with systemic lupus erythematosus" <Biochem.Biophys.REs.Commun>___ー.174. 323-330 (1991)

[Publications] H.Ohishi: "Interaction between the leftーhanded ZーDNA and polyamine:The crystal structure of the d(CG)_3 and Nー(2ーaminoethyl)ー1,4ーdiaminobutane complex" <FEBS Lett>___ー.,. (1991)

[Publications] M.Mizutani: "Negative supercoling of DNA facilitates an interaction between transcription factor IID and the fibroin gene promotor" <Proc.Natl.Acad.Sci>___ー.(USA). 88. (1991)

[Publications] T.Horii: "Mapping of functional domains by truncated by RecA proteins" <J.Mol.Biol>___ー.(1991)

[Publications] Tateishi: "Cーterminal truncated <E.coli>___ー RecA protein,RecA_<5327>,has enhanced DNA binding affinities to singleーand doubleーstranded DNAs:Reguratory role of Cーterminal region in DNA binding" <J.Mol.Biol>___ー.(1991)

[Publications] 箱嶋敏雄、池原森男 (分担執筆): "分子生物の進歩第3巻"蛋白質の機能構造" リボヌクレア-ゼの機能構造" 丸善(日本分子生物学会編), 31 (1990)

[Publications] 箱嶋敏雄、冨田研一 (分担執筆): "<新生化学実験講座第2巻"核酸II構造と性質">___ー X線回折" 東京化学同人(日本生化学会編), (1991)

[Publications] 小川智子 (分担執筆): "<新生化学実験講座核酸の化学I>___ー密度勾配法" 東京化学同人(生化学会編), (1991)

1990 Fiscal Year Annual Research Report

蛋白質/DNA複合体の構造

Principal Investigator

箱嶋 敏雄 大阪大学, 薬学部, 助手 (00164773)

Research Products

[Publications] T.Hakoshima: "Crystallization and preliminary Xーray investigation of nonspecific complexes of a mutant ribonuclease T_1 (Y45W) with 2'AMP and 2'UMP" <J.Mol.Biol.>___ー. 216. 497-499 (1990)

[Publications] T.Hakoshima: "Crystallographic characterization of the mutant and wildーtype ribonuclease T_1 complexed with sevaral ribonucleotides" <J.Biochem (Tokyo)>___ー. 108. 695-698 (1990)

[Publications] S.Nishikawa: "A thermostable mutant of ribonuclease T_1 having three disulfide bond" <Protein Engineering>___ー. 3. 443-448 (1990)

[Publications] 箱嶋 敏雄: "遺伝子発現調節に関与する蛋白質の構造研究" 日本結晶学会誌. 32. 311-314 (1990)

[Publications] M.Kojima: "Interaction of RecA protein with pBR322 DNA modified by Nーhydroxyー2ーacetlyaminofluorene and 4ーhydroxyaminoquinoline 1ーoxide" <Nucle.Acids Res.>___ー. 18. 2707-2714 (1990)

[Publications] A.Higashitani: "ATPーIndependent strand transfer protein from murine spermatocytes,spermatids,and spermatozoa" <Experimental Cell Res.>___ー. 186. 317-323 (1990)

[Publications] H.Ogawa: "Regulation in repressor interaction by RecA protein" <Adv.Biophys.>___ー. 26. 33-48 (1990)

[Publications] 小川 智子: "大腸菌SOS応答反応の分子機構" 蛋白質 核酸 酵素. 35. 893-904 (1990)

[Publications] N.Sakabe: "Protein crystallography using imaging plates and synchrotoron radiation in the photon factory" <Acta Crystallogr.>___ー. A46. C8 (1990)

[Publications] N.Sakabe: "Crystal structure analysis of chicken gizzard Gーactin DNase I complex at 2.4A^^° resolution" <Acta Crystallogr.>___ー. A46. C90 (1990)

[Publications] N.Watanabe: "Crystal structure analysis of ωーamino acid:pyruvata aminotransferase and observation of change in diffraction by time resolution Laue method" <Acta Crystallogr.>___ー. A46. C19 (1990)

[Publications] I.Tanaka: "An automatic diffraction data collection system with an imaging plate" <J.Appl.Crystallogr.>___ー. 23. 334-339 (1990)

[Publications] 田中 勲: "DNA結合蛋白質HU立体構造" 生物物理. 29. 6-11 (1990)

[Publications] T.Tsukihara: "Symmetry and subunit arrengement of Tobacco necrosis virus (TNV)" <Acta Crystallogr.>___ー. B46. 855-860 (1990)

[Publications] T.Ohta: "Purification of DNA supercoiling factor from the posterior silk gland of <Bombyx mori>___ー" <Proc.Natl.Acad.Sci.USA>___ー. 87. 5307-5311 (1990)

[Publications] T.Tamura: "Core promoter of the mouse myelin basic protein gene governs brainーspecific transcription <in vitro>___ー" <EMBO J.>___ー. 9. 3101-3108 (1990)

[Publications] S.Hirose: "DNA supercoling and eukaryotic transcriptionーCause and Effect" <Cell structure and Function>___ー. 15. 133-135 (1990)

[Publications] K.Katayanagi: "Threeーdimensional structure of ribonuclease H from <E.coli>___ー" <Nature>___ー. 347. 306-309 (1990)

[Publications] H.Ueda: "Identification and purification of a <Bombyx mori>___ー homologue of FTZーF1" <Nucle.Acids Res.>___ー. 18. 7229-7234 (1990)

[Publications] K.Wilson: "The termostability of DNAーbinding protein HU from <Bacilli>___ー" <Protein Eng.>___ー. 4. 11-22 (1990)

[Publications] K.Okada: "Negative supercoling of DNA by eukaryotic DNA topoisomerase II and dextran sulfate" <J.Biochem.>___ー. 109. 365-369 (1991)

[Publications] K.Terada: "Demonstration of extrinsic DNA from immune complexes in plasma of a patient with systemic lupus erythematosus" <Biochem.Biophys.REs.Commun>___ー.174. 323-330 (1991)

[Publications] H.Ohishi: "Interaction between the leftーhanded ZーDNA and polyamine:The crystal structure of the d(CG)_3 and Nー(2ーaminoethyl)ー1,4ーdiaminobutane complex" <FEBS Lett>___ー.,. (1991)

[Publications] M.Mizutani: "Negative supercoling of DNA facilitates an interaction between transcription factor IID and the fibroin gene promotor" <Proc.Natl.Acad.Sci>___ー.(USA). 88. (1991)

[Publications] T.Horii: "Mapping of functional domains by truncated by RecA proteins" <J.Mol.Biol>___ー.(1991)

[Publications] Tateishi: "Cーterminal truncated <E.coli>___ー RecA protein,RecA_<5327>,has enhanced DNA binding affinities to singleーand doubleーstranded DNAs:Reguratory role of Cーterminal region in DNA binding" <J.Mol.Biol>___ー.(1991)

[Publications] 箱嶋 敏雄、池原 森男 (分担執筆): "分子生物の進歩 第3巻"蛋白質の機能構造" リボヌクレア-ゼの機能構造" 丸善(日本分子生物学会編), 31 (1990)

[Publications] 箱嶋 敏雄、冨田 研一 (分担執筆): "<新生化学実験講座 第2巻"核酸II構造と性質">___ー X線回折" 東京化学同人(日本生化学会編), (1991)

[Publications] 小川 智子 (分担執筆): "<新生化学実験講座 核酸の化学I>___ー 密度勾配法" 東京化学同人(生化学会編), (1991)

箱嶋敏雄大阪大学, 薬学部, 助手 (00164773)

[Publications] 箱嶋敏雄: "遺伝子発現調節に関与する蛋白質の構造研究" 日本結晶学会誌. 32. 311-314 (1990)

[Publications] 小川智子: "大腸菌SOS応答反応の分子機構" 蛋白質核酸酵素. 35. 893-904 (1990)

[Publications] 田中勲: "DNA結合蛋白質HU立体構造" 生物物理. 29. 6-11 (1990)

[Publications] T.Ohta: "Purification of DNA supercoiling factor from the posterior silk gland of <Bombyx mori>_ー" <Proc.Natl.Acad.Sci.USA>_ー. 87. 5307-5311 (1990)

[Publications] T.Tamura: "Core promoter of the mouse myelin basic protein gene governs brainーspecific transcription <in vitro>_ー" <EMBO J.>_ー. 9. 3101-3108 (1990)

[Publications] K.Katayanagi: "Threeーdimensional structure of ribonuclease H from <E.coli>_ー" <Nature>_ー. 347. 306-309 (1990)

[Publications] H.Ueda: "Identification and purification of a <Bombyx mori>_ー homologue of FTZーF1" <Nucle.Acids Res.>_ー. 18. 7229-7234 (1990)

[Publications] K.Wilson: "The termostability of DNAーbinding protein HU from <Bacilli>_ー" <Protein Eng.>_ー. 4. 11-22 (1990)

[Publications] 箱嶋敏雄、池原森男 (分担執筆): "分子生物の進歩第3巻"蛋白質の機能構造" リボヌクレア-ゼの機能構造" 丸善(日本分子生物学会編), 31 (1990)

[Publications] 箱嶋敏雄、冨田研一 (分担執筆): "<新生化学実験講座第2巻"核酸II構造と性質">___ー X線回折" 東京化学同人(日本生化学会編), (1991)

[Publications] 小川智子 (分担執筆): "<新生化学実験講座核酸の化学I>___ー密度勾配法" 東京化学同人(生化学会編), (1991)