RNA情報機能の分子基盤

1998 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): 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) ; 松藤 千弥 東京慈恵会医科大学, 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・理学研究科・, 教授 (20025662)
• 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10年度の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) <リコーディング>動物細胞アンチザイムのフレームシフト信号の詳細な機能特性とポリアミンの促進作用点の同定、ならびにフレームシフトを利用した新たな翻訳終検定系を作成した。(2) <翻訳終結>解離因子アンチコドン残基5アミノ酸を同定した。さらに、フットプリント分析によりそれらのリボソーム上のトポロジーを明らかにした。(3) <発現調節>テトラヒメナのUAG/UAAコドンを認識するグルタミンtRNA合成酵素を同定した。
2. mRNAプロセシング機構の研究:(1) ゼブラフィッシュのEtrタンパク質の認識するRNA配列をSELEX法により同定した。ならびに、KHモチーフ型RNA結合タンパク質の新規遺伝子xQk6を同定した。(2) 転写と翻訳が連動するRho非依存的ターミネーターでは有為なリードスルーが転写終結点をこえて進行しプロセスされて成熟型のmRNAとなることを明らかにした。
3. tRNA情報機能の研究:(1) Sxl及びHuCタンパク質のRNA結合ドメインの構造をX線結晶あるいはNMR法により解明し、1本鎖RNAの認識機構を構造レベルで解明した。(2) 高度好熱菌の解離因子eRFl結晶を作成した。X線回折データから空間群はP3_1またはP3_2、格子定数がa=b=64.9,c=86.9Åであり、非対称単位にRFlが1分子存在することを明らかにした。

Research Products

• [Publications] Nakamura,Y.,Wada,M.: "Molecular pathobiology and antigenic variation of Pneumocystis carinit." Adv.Parasitol.41. 63-107 (1998)
• [Publications] Pel.H.J.,Moffat,J.G.Ito,k.,Nakamura,Y.,Tate,W.P.: "Escherichia coli release factor-3:resolving the paradox of a typical G protein structure and atypical function with guanine nucleotides." RNA. 4. 47-54 (1998)
• [Publications] Nakamura,Y.: "The inajor surface antigen of Pneumocystis carinii." FEMS Immun.Med.Microbiol.22. 67-74 (1998)
• [Publications] Nakamura,Y.: "Molecular microbiology of Pneumocystis carinii." Jps.J.Med.Mycol.39. 91-98 (1998)
• [Publications] Dasgupta,S.,Fernandez,L.,Kameyama,L.,Inada,T.,Nakamura,Y.,Pappas,A.,Court,D.: "Genetic uncoupling of the dsRNA-binding and RNA-cleavage activities of the Escherichia coli endonbonuclease RNaseIII-the effect of dsRNA binding on gene expression." Mol.Microbiol.28. 629-640 (1998)
• [Publications] Ito,K.,Ebihara,K.,Nakamura,Y.: "The stretch of C-terminal acidic amino acids of translational release factor eRF1 is a primary binding site for eRF3 of fission yeast." RNA. 4. 958-972 (1998)
• [Publications] Ito,K.,Uno,M.,Nakamura,Y.: "Single amino acid substitution in prokaryote polypeptide release factor 2 permits it to terminate translation at all three stop codons." Proc.Natl.Acad.Sci.USA. 95. 8165-8169 (1998)
• [Publications] Nakamura,Y.,Ito,K.: "How protein reads the stop codon and terminates translation(Review)." Genes Cells. 3. 263-278 (1998)
• [Publications] Mita,K.,Morimyo,M.,Hongo,E.,Higashi,T.,Sugaya,K.,Ajimura,M.,Ishihara,Y.,Inoue,H.,Sasanuma,S.,Nohata,J.,Kimura,T.,Koike,Y.,Ito,K.,Nakamura,Y.: "The cDNA and intron catalog of ribosomal protein genes of Schizosaccharomyces pombe." Nucl.Acids Res,In Press. (1999)
• [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 (1998)
• [Publications] T.Hanaki,N.Odawara,Y.K.Nakajima,C.Shimizu,K.Nozaki,T.Mizuno,T.Muramatsu,Y.kuchino and H.Yoshikura: "Two different reactions involved in the primer/template-independent polymerization of dATP and dTTP by Taq DNA polymerase." Biochemical Biophysical Research Communications. 244. 210-219 (1998)
• [Publications] K.Mishima,S.Higashiyama,A.Asai,K.Yamaoka,Y.Nagashima,N.Taniguchi,C.Kitanaka,T.Kirino and Y.Kuchino: "Heparin-binding epidermal growth factor-like growth factor stimulates mitogenic signaling and is highly expressed in human malignant gliomas." Acta Neuropathologica. 96. 322-328 (1998)
• [Publications] A.sugiyama,K.Noguchi,C.Kitanaka,N.Katou,F.Tashiro,M.C.Yoshida and Y.Kuchino: "Molecular cloning and chromosomal mapping of mouse intronless myc gene acting as a potent apoptosis inducer." Gene. 226. 273-283 (1999)
• [Publications] S.Chi,C.Kitanaka,K.Noguchi,T.Mochizuki,Y.Nagashima,M.Shirouzu,H.Fujita,M.Yoshida,W.Chen,A.Asai,M.Himeno,S.Yokoyama and Y.Kuchino: "Oncogenic Ras triggers cell suicide through the activation of a caspase-independent cell death program in human cancer cell." Oncogene,in press. (1999)
• [Publications] C.Kitanaka,Y.Kuchino: "New insight into the signaling pathway for programmed cell death'caspase-independent cell death with necrotic morphology" Cell Death & Differentiation,in press. (1999)
• [Publications] Satriano J,Matsufuji S,Murakami Y,Lortie MJ,Schwartz D,Kelly C,Hayashi S and Blantz RC.: "Agmatine suppresses proliferation by frameshift induction of antizyme and attenuation of cellular polyamine levels." J Biol Chem. 273(25). 15313-15316 (1998)
• [Publications] Ivanov IP,Gesteland RF,Matsufuji S and Atkins JF.: "Programmed frameshifting in the synthesis of mammalian antizyme is +1 in mammals,predominantly +1 in fission yeast,but -2 in budding yeast." RNA. 4(10). 1230-1238 (1998)
• [Publications] Shingo Maegawa,Kunio Yasuda,and Kunio Inoue.: "Matemal mRNA localization of zebrafish DAZ-like gene." Mechanisms of Development. 81. 227-230 (1999)
• [Publications] Tagami,H.and Aiba,H.: "A common role of CRP in transcription activation:CRP acts transiently to stimulate events leading to open complex formation at a diverse set of promoters." EMBO J.17. 1759-1767 (1998)
• [Publications] Takahashi,H.,Inada,T.,Postma,P.,and Aiba,H.: "CRP down-regulatesadenylate cyclase activity by reducing the level of phosphorylated IIAGlc,the glucose-specific phosphotransferase protein,in Escherichia coli." Mol.Gen.Genet.259. 317-326 (1998)
• [Publications] Kimata,K.,Inada,T.,Tagami,H.,and Aiba,H.: "A global repressor(Mlc)is involved in glucose induction of the ptsG gene encoding major glucose transporter in Escherichis coli." Mol.Microbiol.29. 1509-1520 (1998)
• [Publications] Hogema,B.,Arents,J.,Bader,R.,Eijkemans,K.,Inada,T.,Aiba,H.Postma,P.: "Inducerexclusion by glucose 6-phosphate in Escherichia coli." Mol.Microbiol.30. 487-498 (1998)
• [Publications] Abe,H.,Abo,T.,and Aiba,H.: "Regulation of intrinsic terminator by translation in Escherichia coli:transcription termination at a distance downstream." Genes to Cells in press.4. (1999)
• [Publications] Tagami,H.and Aiba,H.: "A novel inactive open complex mediated by UP element at Escherichia coli promoter." Proc.Natl.Acad.Sci.USA in press.96. (1999)
• [Publications] Iwahara,J.,T.Kigawa,K.Kitagawa,H.Masumoto,T.Okazaki and S.Yokoyama: "A helix-turn-helix structure unit in human centromere protein B(CENP-B)." EMBO J.17. 827-837 (1998)
• [Publications] Seio,K.,T.Wada,K.Sakamoto,S.Yokoyama,and M.Sekine: "“Chemical synthesis and properties of conformationally fixed diuridne monophosphates as building blocks of the RNA turn motif"" J.Org.Chem.63. 1429-1443 (1998)
• [Publications] Shirouzu,M.,K.Morinaka,S.Koyama,C.-D.Hu,N.Hori-Tamura,T.Okada,K.Kariya,T.Kataoka,A.Kikuchi,and S.Yokoyama: "“Interactions of the amino acid residue at position 31 of the c-Ha-Ras protein with Raf-1 and RalGDS"" J.Biol.Chem.273. 7737-7742 (1998)
• [Publications] Nureki,O.,D.G,Vassylyev,M.Tateno,A.Shimada,T.Nakama,S.Fukai,M.Konno,T.L.Hendrickson,P.shimmel,and S.Yokoyama: "Enzyme Structure withe two catalytic sites for double-sieve selection of substrate" Science. 280. 578-582 (1998)
• [Publications] Atkins,J.F.,Bock A.,Matsufuji,S.and Gesteland,R.F.: "Dynamics of the Genetic Code" Cold Spring Harbor Laboratory Press., 709 (1998)