Activation of dormant ability by modulating RNA polymerase and global clarification of transcription mechanism

• Project/Area Number: 17380058
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: National Agricultural Research Organization (2006-2008); National Food Research Institute (2005)
• Principal Investigator: 越智 幸三 National Agricultural Research Organization, 食品総合研究所食品バイオテクノロジー研究領域生物機能解析ユニット, ユニット長 (70353985)
• Project Period (FY): 2005 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥15,700,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥2,100,000); Fiscal Year 2008: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2007: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2006: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2005: ¥3,100,000 (Direct Cost: ¥3,100,000)
• Keywords: 応用微生物 / 発酵 / 遺伝子 / RNAポリメラーゼ / 転写制御 / 二次代謝 / 抗生物質 / 緊縮制御 / 放線菌 / 枯草菌 / Thermus / ppGpp / Thermus thermophilus

Research Abstract

微生物における潜在機能を活性化するための技術開発と理論構築を目的として研究を展開し、結果としてRNAポリメラーゼの改変とリボゾームの改変が有効であることを見い出した。特にRNAポリメラーゼの改変は「休眠遺伝子」の活性化においても有効で、本技術は新物質の探索にひとつの有効な手法となるものと思われる。休眠遺伝子の発現原理についても明らかにした。

Research Products

• [Journal Article] Antibacterial discovery in actinomycetes strains with mutations in RNA polymerase or ribosomal protein S12. (2009)
  • Author(s): Hosaka, T., M. Ohnishi-Kameyama, H. Muramatsu, K. Murakami, Y. Tsurumi, S. Kodani, M. Yoshida, A. Fujie, K. Ochi
  • Journal Title: Nature Biotechnology 27 Pages: 462-464
  • Peer Reviewed
• [Journal Article] Identification and Characterization of a Novel Multidrug Resistance Operon (2009)
  • Author(s): Kim, J. Y. T. Inaoka, K. Hirooka, H. Matsuoka, M. Murata, R. Ohki, Y. Adachi, Y. Fujita, K. Ochi.
  • Journal Title: mdtRP (yusOP) of Bacillus subtilis. 191 Pages: 3273-3281
  • Peer Reviewed
• [Journal Article] Bacterial alarmone, guanosine 5'-diphosphate 3'-diphosphate (ppGpp), binds the b' subunit of plastid-encoded plastid RNA polymerase in chloroplasts. (2009)
  • Author(s): Sato, M., K. Takahashi, Y. Ochiai, T. Hosaka, K. Ochi, K. Nabeta.
  • Journal Title: ChemBioChem. 10 Pages: 1227-1233
  • Peer Reviewed
• [Journal Article] Biosynthesis of actinorhodin and related antibiotics : Discovery of alternative routes for quinine formation encoded in the Streptomyces coelicolor gene cluster. (2009)
  • Author(s): Okamoto, S., T. Taguchi, K. Ochi, K. Ichinose.
  • Journal Title: Chem. Biol. 16 Pages: 226-236
  • Peer Reviewed
• [Journal Article] A novel insertion mutation in Streptomyces coelicolor ribosomal S12 protein results in paromomycin resistance and antibiotic over production. (2009)
  • Author(s): Wang, G., T. Inaoka, S. Okamoto, K. Ochi.
  • Journal Title: Antimicrob. Agents Chem other 53 Pages: 1019-1026
  • Peer Reviewed
• [Journal Article] Activation of dormant genes by Nonomuraea ATCC 39727 "mutant type" RNA polymerase. (2009)
  • Author(s): Tala, A., G. Wang,. M. Zemanova., S. Okamoto., K. Ochi., P. Alifano.
  • Journal Title: J. Bacteriol. 191 Pages: 805-814
  • Peer Reviewed
• [Journal Article] Inactivation of KsgA, 16S rRNA methyltransferase, causes vigorous emergence of mutants with high-level kasugamycin resistance. (2009)
  • Author(s): Ochi, K., J-Y. Kim., Y. Tanaka, G. Wang, K. Masuda, H. Nanamiya, S. Okamoto, S. Tokuyasu, Y. Adachi, F. Kawamura.
  • Journal Title: Antimicrob. Agents Chemother. 53 Pages: 193-201
  • Peer Reviewed
• [Journal Article] Novel gene regulation mediated by overproduction of secondary metabolite neotrehalosadiamine in Bacillus subtilis (2009)
  • Author(s): T Inaoka, T Satomura, Y Fujita, K Ochi
  • Journal Title: FEMS Microbiol Lett 291 Pages: 151-156
  • Peer Reviewed
• [Journal Article] Inactivation of KsgA, a 16S rRNA Methyltransferase, Causes Vigorous Emergence of Mutants with High-Level Kasugamycin Resistance (2009)
  • Author(s): K Ochi, J Kim, Y Tanaka, G Wang, K Masuda, H Nanamiya, S Okamoto, S Tokuyama, Y Adachi, F Kawamura
  • Journal Title: Antimicrobial Agents and Chemotherapy 53 Pages: 193-201
  • Peer Reviewed
• [Journal Article] Biosynthesis of Actinorhodin and Related Antibiotics : Discovery of Alternative Routes for quinine Formation Encoded in the act Gene Cluster (2009)
  • Author(s): S Okamoto, T Taguchi, K Ochi, K Ichinose
  • Journal Title: Chemistry & Biology 16 Pages: 226-237
  • Peer Reviewed
• [Journal Article] Dramatic activation of antibiotic production in Streptomyces coelicolor by cumulative drug-resistance mutations. (2008)
  • Author(s): Wang, G., T. Hosaka, K. Ochi.
  • Journal Title: Appl. Environ. Microbiol. 74 Pages: 2834-2840
  • Peer Reviewed
• [Journal Article] Actinoperylone, a novel peryle nequinone-type shunt product, from a deletion mutant of the actVA-OFR5 gene for actinorhodin biosynthesis in Streptomyces coelicolor A3(2). (2008)
  • Author(s): Taguchi, T., S. Okamoto., T. Itoh., Y. Ebizuka., K. Ochi, K. Ichinose.
  • Journal Title: Tetrahedron Letters. 49 Pages: 1208-1211
  • Peer Reviewed
• [Journal Article] Dramatic Activation of Antibiotic Production in Streptomyces coelicolor by Cumulative Drug Resistance Mutations (2008)
  • Author(s): G Wang, T Hosaka, K Ochi
  • Journal Title: Applied and Environmental Microbiology 74 Pages: 2834-2840
  • Peer Reviewed
• [Journal Article] Activation of Dormant Bacterial Genes by Nonomuraea sp. Strain ATCC 39727 Mutant-Type RNA Polymerase (2008)
  • Author(s): A Tala, G Wang, M Zemanova, S Okamoto, K Ochi, P Alifano
  • Journal Title: Journal of Bacteriology 191 Pages: 805-814
  • Peer Reviewed
• [Journal Article] A Novel Insertion Mutation in Streptomyces coelicolor Ribosomal S12 protein results in paromomycin Resistance and Antibiotic Overproduction (2008)
  • Author(s): G Wang, T Inaoka, S Okamoto, K Ochi
  • Journal Title: Antimicrobial Agents and Chemotherapy 53 Pages: 1019-1026
  • Peer Reviewed
• [Journal Article] Dramatic activation of antibiotic production in Streptomyces coelicolor by cumulative drug resistance mutations (2008)
  • Author(s): Goujun Wang, et. al.
  • Journal Title: Applied and Environmental Microbiology 74
  • Peer Reviewed
• [Journal Article] Identification of the RsmG methyltransferase target as 16S rRNA nucleotide G527 and characterization of Bacillus subtilis rsmG mutants. (2007)
  • Author(s): Nishimura, K., S. K. Johansen., T. Inaoka., T. Hosaka., S. Tokyuyama., T. Tahara., S. Okamoto., F. Kawamura., S. Douthwaite., K. Ochi.
  • Journal Title: J. Bacteriol. 189 Pages: 6068-6073
  • Peer Reviewed
• [Journal Article] The rare earth, scandium, causes antibiotic overproduction in Streptomyces spp. (2007)
  • Author(s): Kawai, K., G. Wang., S. Okamoto., K. Ochi.
  • Journal Title: FEMS Microbiol. Lett. 274 Pages: 311-315
  • Peer Reviewed
• [Journal Article] Loss of a conserved 7-methylguanosine modification in 16S rRNA confers low-level streptomycin resistance in bacteria. (2007)
  • Author(s): Okamoto, S., A. Tamaru., C. Nakajima., K. Nishimura., Y. Tanaka., S. Tokuyama., Y. Suzuki., K. Ochi.
  • Journal Title: Mol. Microbiol. 63 Pages: 1096-1106
  • Peer Reviewed
• [Journal Article] Isolation and characterization of soprulation-initiation mutation in Bacillus subtilis prfB gene. (2007)
  • Author(s): Asai, K., T. Inaoka., H, Nanamiya., Y. Sadaie., K. Ochi., F. Kawamura.
  • Journal Title: Biosci. Biotechnol. Biochem. 71 Pages: 397-406
  • Peer Reviewed
• [Journal Article] Mutations in rsmG, encoding a 16S rRNA methyltransferase, result in low-level streptomycin resistance and antibiotic overproduction. (2007)
  • Author(s): Nishimura, K., T. Hosaka., S. Tokuyama., S. Okamoto., K. Ochi.
  • Journal Title: J. Bacteriol. 189 Pages: 3876-3883
  • Peer Reviewed
• [Journal Article] Glucose up take pathway-specific regulation of synhtesis of neotrehalosadiamine, an novel autoinducer produced in Bacillus subtilis. (2007)
  • Author(s): Inaoka, T., K. Ochi.
  • Journal Title: J. Bacteriol. 189 Pages: 65-75
  • Peer Reviewed
• [Journal Article] A fail-safe system for the ribosome under zinc-limiting conditions in Bacillus subtilis. (2007)
  • Author(s): Natori, Y., H. Nanamiya., G. Akanuma., S. Kosono., T. Kudo., K. Ochi., F. Kawamura.
  • Journal Title: Mol. Microbiol. 63 Pages: 294-307
  • Peer Reviewed
• [Journal Article] Identification of the RsmG Methyltransferase target as 16S rRNA nucleotide G527 and characterization of Bacillus subtilis rsmG mutants (2007)
  • Author(s): Kenji Nishimura, et. al.
  • Journal Title: Journal of Bacteriology 189 Pages: 6068-6073
  • Peer Reviewed
• [Journal Article] The rare earth,scandium,causes antibiotic overproduction in Streptomyces spp. (2007)
  • Author(s): Keiichi Kawai, et. al.
  • Journal Title: FEMS Microbiology Letters 274 Pages: 311-315
  • Peer Reviewed
• [Journal Article] A fail-safe system for the ribosome under zinc-limiting conditions in Bacillus subtilis (2007)
  • Author(s): Yousuke Natori et al.
  • Journal Title: Molecular Microbiology 63 Pages: 294-307
• [Journal Article] Glucose uptake pathway-specific regulation of synthesis of meotrehalosadiamine, a novel autoinducer produced in bacillus subtilis (2007)
  • Author(s): Takashi Inaoka et al.
  • Journal Title: Journal of Bacteriology 189 Pages: 65-75
• [Journal Article] Loss of a conserved 7-methylguanosine modification in 16S rRNA confers low-level streptomycin resistance in bacteria (2007)
  • Author(s): Susumu Okamoto et al.
  • Journal Title: Molecular Microbiology 63 Pages: 1096-1106
• [Journal Article] Increased expression of ribosome recycling factor is responsible for the enhanced protein synthesis during the late growth phase in an antibitoic-overproducing Streptomyces coel icolor ribosomal rpsL mutant. (2006)
  • Author(s): Hosaka, T., J. Xu., K. Ochi.
  • Journal Title: Mol. Microbiol. 61 Pages: 833-897
  • Peer Reviewed
• [Journal Article] Physiological analysis of tehstringent response elicited in an extreme thermophilic bacterium, The rmus thermophilus. (2006)
  • Author(s): Kasai, K., T. Nishizawa., K. Takahashi., T. Hosaka., H. Aoki., K. Ochi.
  • Journal Title: J. Bacteriol. 188 Pages: 7111-7122
  • Peer Reviewed
• [Journal Article] EshA accentuates ppGpp accumulation and is conditionally required for antibiotic producton in Streptomyces coelicolor A3(2). (2006)
  • Author(s): Saito, N., J. Xu., T. Hosaka., S. Okamoto., H. Aoki., M. J. Bibb., K. Ochi.
  • Journal Title: J. Bacteriol. 188 Pages: 4952-4961
  • Peer Reviewed
• [Journal Article] Improvement of α-amylase production by modulating the ribosomal component S12 protein in Bacillus subtilis 168. (2006)
  • Author(s): Kurosawa, K., T. Hosaka, N. Tamehiro, T. Inaoka, K. Ochi.
  • Journal Title: Appl. Environ. Microbiol. 72 Pages: 71-77
  • Peer Reviewed
• [Journal Article] Improvement of α-amylase production by modulation of ribosomal component protein S12 in Bacillus subtilis 168 (2006)
  • Author(s): Kazuhiko Kurosawa et al.
  • Journal Title: Applied and Environmental Microbiology 72 Pages: 71-77
• [Journal Article] EshA accentuates ppGpp accumulation and is conditionally required for antibiotic production in Streptomyces coelicolor A3(2) (2006)
  • Author(s): Natsumi Saito et al.
  • Journal Title: Journal of Bacteriology 188 Pages: 4952-4961
• [Journal Article] Increased expression of ribosome recycling factor is responsible for the enhanced protein synthesis during the late growth phase in an antibiotic-overproducing Streptomyces coelicolor ribosomal rpsL mutant (2006)
  • Author(s): Takeshi Hosaka et al.
  • Journal Title: Molecular Microbiology 61 Pages: 883-897
• [Journal Article] Physiological analysis of the stringent response elicited in an extreme thermophilic bacterium, Thermus thermophilus (2006)
  • Author(s): Koji Kasai et al.
  • Journal Title: Journal of Bacteriology 188 Pages: 7111-7122
• [Journal Article] Improvement of α-amylase production by modulation of ribosomal component protein S12 in Bacillus subtilis 168 (2006)
  • Author(s): Kazuhiko Kurosawa
  • Journal Title: Applied and Environmental Microbiology 72・1 Pages: 71-77
• [Journal Article] Possible involvement of ActVI-ORFA in transcriptional regulation of actVI tailoring-step genes for actinorhodin biosynthesis.
  • Author(s): Taguchi, T., S. Okamoto., A. Lezhava., A. Li., K. Ochi., Y. Ebizaka., K. Ichinose.
  • Journal Title: FEMS Microbiol. Lett. 269 Pages: 234-239
  • Peer Reviewed
• [Presentation] ストレプトマイシン弱耐性変異rsmG導入による Streptomyces 属の二次代謝能の変化 (2008)
  • Author(s): 田中幸徳, 他
  • Organizer: 2008年度日本放線菌学会
  • Place of Presentation: 山梨県山梨市
  • Year and Date: 2008-10-10
• [Presentation] Actinorhodin 生合成におけるActVA-5, ActVA-6 の機能解析 (2008)
  • Author(s): 田口貴章、岡本晋、越智幸三、市瀬浩志
  • Organizer: 日本放線菌学会
  • Place of Presentation: フルーツパーク富士屋ホテル(山梨県山梨市)
• [Presentation] ストレプトマイシン弱耐性変異rsmG導入によるStreptomyces属の二次代謝能の変化 (2008)
  • Author(s): 田中幸徳、岡本晋、小松譲、徳山真治、池田治生、越智幸三
  • Organizer: 日本放線菌学会
  • Place of Presentation: フルーツパーク富士屋ホテル(山梨県山梨市)
• [Presentation] Streptromyces coelicolor変異株における染色体環状化の解析 (2008)
  • Author(s): 西川友也、荒川賢治、王国君、越智幸三、木梨陽康
  • Organizer: 日本放線菌学会
  • Place of Presentation: フルーツパーク富士屋ホテル(山梨県山梨市)
• [Presentation] 希土類元素「スカンジウム」による放線菌二次代謝の活性化 (2007)
  • Author(s): Kozo Ochi, et. al.
  • Organizer: 2007年度日本放線菌学会大会
  • Place of Presentation: しまなみ交流館(テアトロシェルネ)・広島県尾道市
  • Year and Date: 2007-06-01
• [Book] In Gloval Regulatory Networks in Bacillus subtilis (2007)
  • Author(s): Regulation of secondary metabolism in Bacillus subtilis
  • Publisher: Transworld Research Network
• [Book] 微生物の二次代謝に生理的意義はあるのか? 抗生物質生産の新たな意味を探る、化学と生物 (2007)
  • Author(s): 越智幸三、稲岡隆史
• [Book] From microbial differentiation to ribosome engineering. Biosci. Biotechnol. Biochem. (2007)
  • Author(s): Ochi, K.
• [Book] 微生物の「休眠遺伝子」を活性化し抗生物質などを作り出す技術を開発、ブレインテクノニュース (2007)
  • Author(s): 越智幸三
• [Book] リボゾーム工学による微生物からの有用物質発見、月刊バイオインダストリー2007年10月号 (2007)
  • Author(s): 保坂毅、王国君、岡本晋、越智幸三
• [Book] Global regulatory networks in Bacillus subtilis (2007)
  • Author(s): Takashi Inaoka, et. al.
  • Total Pages: 249
  • Publisher: Transworld Research Network
• [Book] 蛋白質 核酸 酵素 (2005)
  • Author(s): 戸澤 譲, 越智 幸三
  • Total Pages: 1
  • Publisher: 共立出版株式会社
• [Book] THE WAKSMAN FOUNDATION OF JAPAN INC. Report of Researchs in 2004 (2005)
  • Author(s): Kozo Ocni
  • Total Pages: 12
  • Publisher: THE WAKSMAN FOUNDATION OF JAPAN INC.