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1 Montiel, M.D., Krzewinski-Recchi, M.A., Delannoy, P., & Harduin-Lepers, A.Biochem. J.,,. Nagata, Y., Yamashiro, S., Yodoi, J., Lloyd, K.O., Shiku, H., & Furukawa, K.J. Biol. Chem.,,. Dohi, T., Nishikawa, A., Ishizuka, I., Totani, M., Yamaguchi, K., Nakagawa, K., Saitoh, O., Ohshiba, S., & Oshima, M. Biochem. J.,,. Morton, J.A., Pickles, M.M., & Vanhegan, R.I.Immunol. Invest.,,. Dohi, T., Ohta, S., Hanai, N., Yamaguchi, K., & Oshima, M. J. Biol. Chem.,,. Dohi, T., Hanai, N., Yamaguchi, K., & Oshima, M.J. Biol. Chem.,,. Kawamura, Y.I., Kawashima, R., Fukunaga, R., Hirai, K., Toyama-Sorimachi, N., Tokuhara, M., Shimizu, T., & Dohi, T. Cancer Res.,,. Morton, J.A., Pickles, M.M., & Vanhegan, R.I.Immunol. Invest.,,. Malagolini, N., Dall Olio, F., Di Stefano, G., Minni, F., Marrano, D., & Serafini-Cessi, F.Cancer Res.,,. Piller, F., Cartron, J-P., & Tuppy, H.Blood Transf. Immunohaematol.,,. Capon, C., Maes, E., Michalski, J.C., Leffler, H., & Kim, Y.S. Biochem. J.,,. Dohi, T., Yuyama, Y., Natori, Y., Smith, P.L., Lowe, J.B., & Oshima, M.Int. J. Cancer,,. Kawamura, Y.I., Toyota, M., Kawashima, R., Hagiwara, T., Suzuki, H., Imai, K., Shinomura, Y., Tokino, T., Kannagi, R., & Dohi, T.Gastroenterology, in press. Miyazaki, K., Ohmori, K., Izawa, M., Koike, T., Kumamoto, K., Furukawa, K., Ando, T., Kiso, M., Yamaji, T., Hashimoto, Y., Suzuki, A., Yoshida, A., Takeuchi, M., & Kannagi, R. Cancer Res.,,. Dall Olio, F., Malagolini, N., Di Stefano, G., Ciambella, M., & Serafini-Cessi, F.Biochem. J.,,. Sd a blood group carbohydrate antigen, exclusively expressed in normal gastrointestinal mucosa but not in cancer tissues Yuki I. Kawamura Department of Gastroenterology, Research Institute, International Medical Center of Japan, Toyama, Shinjuku-ku, Tokyo, Japan CCA RNA trna CCA DNA RNA RNA RNA mrna mrna RNA trna mrna trna mrna trna trna L trna trna CCA C -C -A trna trna CCA CCA RNA trna CCA trna CCA CCA RNA trna CCA CCA trna RNA RNase PH

2 CCA trna CCA CCA RNA CCA A PAP mrna TdT DNA dntp DNA b pol b KNT pol b DNA CCA PAP TdT DNA PAP TdT d NTP CCA CCA RNA CCA C C A trna CCA CCA CCA trna CCA CCA CCA II CCA I A mrna I A II Bacillus stearothermophilus CCA A Archaeglobus fulgidus CCA A II CCA I CCA I CCA A ATP CTP I II I II CCA I CCA II CCA N ATP CTP CCA C C A CCA C C A CCA trna ATP CCA DNA trna CCA I CCA I Archaeglobus fulgidus CCA trna NTP A trna TΨC trna CCA mini-d trna D mini-dc C mini-dc CTP mini-

3 DCC C mini-dcc ATP mini-dcca A Steitz Nature A. fulgidus CCA trna A release Steitz CCA I CCA head neck body tail a trna TΨC tail trna b tail CCA trna RNA RNA trna neck body d trna P S S N T c mini-d D A head catalytic triad Glu Asp Asp S mini-dc A S C S trna CCA mini-d mini-dc trna A S C A C S mini-dc S C Glu AlaHis Ser His Ala CMP AMP C CTP mini-dc CTP head neck Glu C CTP head catalytic triad CTP C RNA CMP head knock-in CTP C mini-dcc C S C P C C Glu Thr CCA head mini- DC ATP N T ATP mini-dcca trna CCA D Steitz release head D CCA CCA Dhead trna CCA CTP ATP CTP ATP A C CTP ATP head Arg CTP ATP head

4 4 3 2 生化学 第8 0巻 第5号 図1 ウシ PAP クラス I Archaeglobus fulgidus CCA 付加酵素 AfCCA およびクラス II Bacillus stearothermophilus CCA 付加酵素 BstCCA の CCA 付加酵素の結晶構造 の比較 図2 Archaeglobus fulgidus 由来 CCA 付加酵素と trna ミニヘリックスの複合体の 結晶構造 a 全体構造 b tail ドメインによる TΨC ループの認識 c 活性部位におけるプ ライマー末端 新規ヌクレオチドの認識 d body ドメインによるステムの認識 みにれびゆう

5 4 3 3 2 0 0 8年 5月 図4 CTP 付加反応における head ドメインの構造 変化 図5 CCA 付加酵素による重合反応のダイナミクス 図3 クラス I CCA 付加酵素による重合反応における各伸長段 階のスナップショット みにれびゆう

6 Arg ATP CCA CCA Viceanchored knock-in-and-lock trna L CCA tail TΨC D A CTP head knock-in A RNA A C S S TΨC CTP head C CMP C S C P C C N T ATP RNA CCA CCA D A head trna CCA trna tail I CCA A tail mrna A A Sprinzl, M. & Cramer, F.Prog. Nucleic Acid Res. Mol. Biol.,,. Nissen, P., Hansen, J., Ban, N., Moore, P.B., & Steitz, T.A. Science,,. Li, F., Xiong, Y., Wang, J., Cho, H.D., Tomita, K., Weiner, A. M.,&Steitz,T.A.Cell,,. Okabe, M., Tomita, K., Ishitani, R., Ishii, R., Takeuchi, N., Arisaka, F., Nureki, O., & Yokoyama, S.EMBO J.,,. Xiong, Y., Li, F., Wang, J., Weiner, A.M., & Steitz, T.A. Mol. Cell,,. Tomita, K., Fukai, S., Ishitani, R., Ueda, T., Takeuchi, T., Vassylyev, D.G., & Nureki, O.Nature,,. Xiong, Y. & Steitz, T.A.Nature,,. Schimmel, P. & Yang, X.-L.Nat. Struct. Mol. Biol.,,. Tomita, K., Ishitani, R., Fukai, S., & Nureki, O.Nature,,. Dynamic mechanism of CCA-adding polymerization reaction Osamu Nureki Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta-cho, Midori-ku, Yokohama-shi, Kanagawa, Japan C C C C ELOVL C C ALD

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