RNA情報機能の分子基盤

1999 Fiscal Year Annual Research Report

• Project/Area Number: 09278102
• Research Institution: The University of Tokyo
• Principal Investigator: 中村 義一 東京大学, 医科学研究所, 助教授 (40114590)
• Co-Investigator(Kenkyū-buntansha): 松藤 千弥 東京慈恵医科大学, 医学部, 助教授 (50192753) ; 口野 嘉幸 国立がんセンター研究所, 部長(研究職) (60124418) ; 饗場 弘二 名古屋大学, 大学院・理学研究所, 教授 (20025662) ; 横山 茂之 東京大学, 大学院・理学系研究科, 教授 (00159229) ; 井上 邦夫 奈良先端科学技術大学院大学, 助手 (40252415)
• Keywords: RNA / 翻訳調節 / 転写後制御 / リボソーム / 分子擬態 / 終止コドン / mRNA動態 / RNA結合蛋白質

Research Abstract

平成11年度の主要な研究成果を以下に整理して述べる。
1.翻訳調節におけるRNA情報機能の研究:(1)遺伝暗号の中で最後まで解読のメカニズムが明かにされなかった終止コドンの解読機構が解明された。コドン特異的なバクテリア解離因子RF1/RF2の"tripeptides"がtRNAアンチドンと同様に直接終止コドンを識別するペプチド性アンチドンとして働くことが証明された。これは歴史的な発見である。(2)高等動物での試験管内の翻訳終結反応を正確にモニターできる系が作成された。これはフレームシフトと終結反応との競合を利用したアッセイ系で、真核系での精密な翻訳終結とリコーディング機構の解析に突破口を開いた。
2.RNAプロセシング機構の研究:(1)大腸菌tmRNAの標的を検索した結果、野生株においてtmRNA由来のタグが付加されたポリペプチドが多数存在することが明かにされた。これは生理的な条件下でtmRNAによるタグ付加を示した初めての実証である。(2)脊椎動物において神経特異的に発現するRNA結合蛋白質Huが細胞内において核と細胞質の間をシャトルしていることが示唆されてきたが、そのシャトリングに必要な領域を特定し、変異体を利用した解析からHu蛋白質が神経細胞の分化に重要な役割を担っていることが明かにされた。
3.RNA情報構造機能の研究:ショウジョウバエの性決定に係わるSxl蛋白質とその認識配列RNAとの複合体の結晶構造が解かれた。二つのRNA結合ドメインが認識配列を特異的に認識するその仕組は、これまで知られていないRNA/蛋白質の分子認識のメカニズムを明かにしたもので重要な成果である。

Research Products

• [Publications] Crawford, D.-J. G., Ito, K., Nakamura, Y., Tate, W. P.: "Indirect regulation of termination efficiency at highly expressed genes and recoding sites by the factor recycling function of Escherichia coli release factor RF3"EMBO J.. 18. 727-732 (1999)
• [Publications] Ebihara, K., Nakamura, Y.: "C-terminal interaction of translational release factors eRF1 and eRF3 of fission yeast : G-domain uncoupled binding and the role of conserved amino acids"RNA. 5. 739-750 (1999)
• [Publications] Fujiwara, T., Ito, K., Nakayashiki, T., Nakamura, Y.: "Amber mutations in ribosome recycling factors of Escherichia coli and Thermus thermophilus : evidence for C-terminal modulator element"FEBS Lett.. 447. 297-302 (1999297)
• [Publications] Yoshimura, K., Ito, K., Nakamura, Y.: "Amber (UAG) suppressors affected in UGA/UAA-specific polypeptide release factor 2 of bacteria : genetic prediction of initial binding to ribosome preceding stop codon recognition"Genes Cells. 4. 253-266 (1999)
• [Publications] Powell, B., Peters III, H. K., Nakamura, Y., Court, D.: "Cloning and analysis of the rnc-era-recO operon from Pseudomonas aeruginosa"J. Bacteriol.. 181. 5111-5113 (1999)
• [Publications] Karamyshev, A., Ito, K., Nakamura, Y.: "Polypeptide release factor eRF1 from Tetrahymena thermophila : cDNA cloning, purification and complex formation with yeast eRF3"FEBS Lett.. 457. 483-488 (1999)
• [Publications] Karamyshev, A., Karamycheva, Z., Ito, K., Matsufuji, S., Nakamura, Y.: "Overexpression and purification of recombinant eRF1 proteins of rabbit and Tetrahymena thermophila"Biochemistry (Moscow). 64. 1648-1658 (1999)
• [Publications] Wada, M., Nakamura, Y.: "Type-II major-surface-glycoprotein family of Pneumocystis carinii"J. Euk. Microbiol.. 46. 151S-152S (1999)
• [Publications] Wada, M., Nakamura, Y.: "Immunological characterization of surface subtilisin-like protease (SSP) of Pneumocystis carinii"J. Euk. Microbiol.. 46. 125S-126S (1999)
• [Publications] Wada, M., Nakamura, Y.: "Type-II major-surface-glycoprotein family of Pneumocystis carinii under the control of novel expression elements"DNA Res.. 6. 211-217 (1999)
• [Publications] Tin, O. F., Rykunova, A. L., Muranova, T. A., Toyoda, T., Ito, K., Suzuki, T., Watanabe, K., Garber, M. B., Nakamura, Y.: "Proteolytic fragmentation of polypeptide release factor 1 of Thermus thermophilus and crystallization of the stable fragments"Biochimie, In Press. (2000)
• [Publications] Tanaka, R., Satoh, H., Moriyama, M., Satoh, K., Morishita, Y., Yoshida, S., Watanabe, T., Nakamura, Y., Mori, S.: "Intronic U50 small-nucleolar-RNA (snoRNA) host gene of no protein-coding potential is mapped at the chromosome breakpoint t(3;6)(q27;q15) of human B-cell lymphoma"Genes Cells, In Press. (2000)
• [Publications] Ito, K., Uno, M., Nakamura, Y.: "A tripeptide 'anticodon deciphers' stop codons in messenger RNA"Nature. 403. 680-683 (200)
• [Publications] Abe, H., Abo, T., Aiba, H.: "Regulation of intrinsic terminator by translation in Escherichia coli : transcription termination at a distance downstream"Genes to Cells. 4. 87-97 (1999)
• [Publications] Tagami, H., Aiba, H.: "An inactive open complex mediated by UP element ant Escherichia coli promoter"Proc. Natl. Acad. Sci. USA. 96. 12914-12919 (1999)
• [Publications] Tanaka, Y., Kimata, K., Inada, T., Tagami, H., Aiba, H.: "Negative regulation of the pts operon by Mls : Mechanism underlying glucose induction in Escherichia coli"Genes to Cells. 4. 391-399 (1999)
• [Publications] Sugiyama, A., Noguchi, K., Kitanaka, C., Katou, N., Tashiro, F., Ono, T., Yoshida, M. C., Kuchino, Y.: "Molecular cloning and chromosomal mapping of mouse intronless myc gene acting as a potent apoptosis inducer"Gene. 226. 273-283 (1999)
• [Publications] Mochizuki, T., Kitanaka, C., Noguchi, K., Muramatsu, T., Asai, A., Kuchino, Y.: "Physical and functional interaction between Pim-1 kinase and Cdc25A phosphatase : implication for the Pim-1-mediated activation of the c-Myc signaling pathway"J. Biol. Chem.. 274. 18659-18666 (1999)
• [Publications] Noguchi, K., Kitanaka, C., Yamana, H., Kokubu, A., Mochizuki, T., Kuchino, Y.: "Regulation of c-Myc through phosphorylation at Ser-62 and -71 by c-Jun N-terminal kinase"J. Biol. Chem.. 274. 32580-32587 (1999)
• [Publications] Watanabe, S., Muramatsu, T., Ao, H., Hirayama, Y., Takahashi, K., Tanokura, M., Kuchino, Y.: "Molecular cloning of the Lon protease gene form Thermus thermophilus HB8 and characterization of its gene product"E. J. Biochem.. 266. 811-819 (1999)
• [Publications] Itoh, S., Hattori, T., Hayashi, H., Mizutani, Y., Todo, M., Takii, T., Yang, D., Lee, J. C., Matsufuji, S., Murakami, Y., Chiba, T., Onozaki, K.: "Antiproliferative effect of IL-1 is mediated by p38 mitogen-activated protein kinase in human melanoma cell A 375"J. Immunol.. 162(12). 7434-7440 (1999)
• [Publications] Murakami, Y., Matsufuji, S., Hayashi, S., Tanahashi, N., Tanaka, K.: "ATP-dependent inactivation and sequestration of ornithine decarboxylase by the 26S proteasome is a prerequisite for degradation"Mol. Cell. Biol.. 19. 7216-7227 (1999)
• [Publications] Suka, M., Matsufuji, S., Murakami, Y.: "Dietary induction of ornithine decarboxylase in male mouse kidney"Biochim. Biophys. Acta.. 1472. 455-461 (1999)
• [Publications] Saito, T., Hascilowicz, T., Ohkido, I., Kikuchi, Y., Okamoto, H., Hayashi, S., Murakami, Y., Matsufuji, S.: "Two zebrafish antizymes with different expression and activities"Biochem. J.. 345. 99-106 (2000)
• [Publications] Isoyama, T., Kamoshita, N., Yasui, K., Iwai, A., Shiroki, K., Toyoda, H., Yamada, A., Takasaki, Y., Nomoto, A.: "Lower concentration of La protein required for internal ribosomal entry onhepatitis C virus RNA than on poliovirus RNA"J. Gen. Virol.. 80(9). 2319-2328 (1999)
• [Publications] Maegawa, S., Yasuda, K., Inoue, K.: "Maternal mRNA localization of zebrafish DAZ-like gene"Mech. Dev.. 81. 223-226 (1999)
• [Publications] Handa, N., Nureki, O., et al.: "Structural bases for recognition of the tra mRNA precursor by the Sex-lethal protein"Nature. 398. 579-585 (1999)
• [Publications] Kigawa, T., et al.: "Cell-free production and stable Oisotope labeling of milligram-quantities of proteins"FEBS Lett.. 442. 15-19 (1999)
• [Publications] Chi, S.-W., et al.: "Chemical shift perturbation studies of the Interactions of the second RNA-binding domain of the Drosophila Sex-lethal protein with the transformer pre-mRNA polyuridine tract and 3' splice site sequences"Eur. J. Biochem.. 260. 649-660 (1999)
• [Publications] Chi, S., et al.: "Oncogenic Ras triggers cell suicide through the activation of a caspase-independent cell death program in human cancer cells"Oncogene. 18. 2281-2290 (1999)
• [Publications] Horie, N., et al.: "Modified nucleosides in the first positions of the anticodons of tRNALeu4 and tRNALeu5 from Escherichia coli"Biochemistry. 38. 207-217 (1999)
• [Publications] Atkins, J. F., Bock, A., Matsufuji, S., Gesteland, R. F.: "Dynamics of the Genetic Code"Cold Spring Habor Laboratory Press. 709 (1999)