Development of genetic system of rRNA genes and its use in B. subtilis

• Project/Area Number: 19570006
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Genetics/Genome dynamics
• Research Institution: Rikkyo University
• Principal Investigator: KAWAMURA Fujio Rikkyo University, 理学部, 教授 (10126039)
• Project Period (FY): 2007 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2009: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2008: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2007: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: リボソーム / rRNA / 翻訳 / 枯草菌 / 胞子形成 / rRNAオペロン / 抗生物質耐性

Research Abstract

We deleted the 10 rrn operons of B. subtilis sequentially, and constructed seven mutants, each harboring a single rrn operon in their genome. The growth rates and sporulation frequencies of these mutants were reduced drastically as compared with those of the wild type strain, indicating that normal growth as well as sporulation requires multiple copy numbers of rRNA operon. Further analysis using mutants having various numbers of rRNA operon showed that normal sporulation requies more than 5 copies of rRNA operon.

Research Products

• [Journal Article] Towards an elucidation of the roles of the ribosome during different growth phases in Bacillus subtilis. (2010)
  • Author(s): Nanamiya, H., Kawamura F.
  • Journal Title: Biosci. Biotechnol. Biochem. 74 Pages: 451-461
  • NAID: 10027552065
• [Journal Article] Inhibition of Septation in Bacillus subtilis by a peptide antibiotic (2010)
  • Author(s): Shimotohno, W.K., Kawamura F., Natori, H., Nanamiya, H., Magae, J., Ogata, H., Endo, T., Suzuki, T., Yamaki, H.
  • Journal Title: edein B1. Biol. Pharm. Bull. 33 Pages: 568-571
• [Journal Article] Inhibition of septation in Bacillus subtilis by a peptide antibiotic, edeine B1 (2010)
  • Author(s): Kumiko W.Shimotohno
  • Journal Title: Biological & Pharmaceutical Bulletin 33 Pages: 568-571
  • Peer Reviewed
• [Journal Article] Transcription activity of individual rrn operons in Bacillus subtilis mutants deficient in (p)ppGpp synthetase genes, relA, yjbM, and ywaC. (2009)
  • Author(s): Natori, Y., Tagami, K., Murakami, K., Yoshida, S., Tanigawa, O., Moh, Y., Masuda, K., Wada, T., Suzuki, S., Nanamiya, H., Tozawa, Y., Kawamura, F.
  • Journal Title: J. Bacteriol. 191 Pages: 4555-4561
• [Journal Article] Inactivation of KsgA, a 16S rRNA methyltransferase, causes vigorous emergence of mutants with high-level kasugamycin resistance. (2009)
  • Author(s): Ochi, K., Kim, J-Y., Tanaka, Y., Wang, G., Masuda, K., Nanamiya, H., Okamoto, S., Tokuyama, S., Adachi, Y., Kawamura, F.
  • Journal Title: Antimicrob. Agents Chemother. 53 Pages: 193-201
• [Journal Article] Transcription activity of individual rrn operons in Bacillus subtilis mutants deficient in(p)ppGpp synthetase genes, relA, yjbM, and ywaC. (2009)
  • Author(s): Yousuke Natori
  • Journal Title: Journal of Bacteriology 191 Pages: 4555-4561
  • Peer Reviewed
• [Journal Article] Inactivation of KsgA, a 16S rRNA methyltransferase, causes vigorous emergence of mutqants with high-level kasugamycin resistance (2009)
  • Author(s): Kozo Ochi
  • Journal Title: Antimicrobial Agents and Chemotherapy 53 Pages: 193-201
  • Peer Reviewed
• [Journal Article] Identification and functional analysis of novel (p)ppGpp synthetase genes in Bacillus subtilis. (2008)
  • Author(s): Nanamiya H., Kasai K., Nozawa A., Yun C.-S., Narisawa T., Murakami K., Natori Y., Kawamura F., Tozawa Y.
  • Journal Title: Mol. Microbiol. 67 Pages: 291-304
• [Journal Article] Identification and functional analysis of novel(p)ppGpp synthetase genes in Bacillus subtiblis (2008)
  • Author(s): Nanamiya Hideaki
  • Journal Title: Molecular Microbiology 67 Pages: 291-304
  • Peer Reviewed
• [Journal Article] A fail-safe system for the ribosome under zinc-limiting conditions in Bacillus subtilis. (2007)
  • Author(s): Natori, Y., Nanamiya, H., Akanuma, G., Kosono, S., Kudo, T., Ochi, K., Kawamura, F.
  • Journal Title: Mol. Microbiol. 63 Pages: 294-307
• [Journal Article] Permuted tRNA genes expressed via a circular RNA intermediate in Cyanidioschyzon merolae (2007)
  • Author(s): Soma A., Onodera A., Sugahara J., Kanai A., Yachie N., Tomita M., Kawamura F., Sekine Y.
  • Journal Title: Science 318 Pages: 450-453
• [Journal Article] Model-based definition of population heterogeneity and its effects on metabolism in sporulating Bacillus subtilis. (2007)
  • Author(s): Morohashi M., Ohashi., Tani S., Ishii K., Itaya M., Nanamiya H., Kawamura F., Tomita M., Soga T.
  • Journal Title: J. Biochem. 142 Pages: 183-191
  • NAID: 10020097136
• [Journal Article] Identification of the RsmG methyltransferase target as 16S rRNA nucleotide G527 and characterization of Bacillua subtilis rsmG mutants. (2007)
  • Author(s): Nishimura K., Johansen S.K., Inaoka T., Hosaka T., Tokuyama S., Tahara Y., Okamaoto S., Kawamura F., Douthwaite S., Ochi K.
  • Journal Title: J. Bacteriol. 189 Pages: 6068-6073
• [Journal Article] Isolation and characterization of sporulation-initiation mutation in the Bacillus subtilis prfB gene. Biosci. (2007)
  • Author(s): Asai, K., Inaoka, T., Nanamiya, H., Sadaie, Y., Ochi, K., Kawamura, F.
  • Journal Title: Biotechnol. Biochem. 71 Pages: 397-406
• [Journal Article] Isolation and characterization of sporulation-initiation mutation in theBaeillus subtilis prfB gene (2007)
  • Author(s): Asai Kei
  • Journal Title: Biosci.Biotechnol.Biochem 71 Pages: 397-406
  • Peer Reviewed
• [Journal Article] Identification of the RsmG methyltransferase target as 16S rRNA nucleotide G527 and characterization of Baeillus subtilis rsmG mutants (2007)
  • Author(s): Nishimura Kenji
  • Journal Title: Journal of Bacteriology 189 Pages: 6068-6073
  • Peer Reviewed
• [Journal Article] Model-based definition of population heterogeneity and its effects onmetabolism in sporulating Baeillus subtilis (2007)
  • Author(s): Morohashi Mineo
  • Journal Title: Journal of Biochemistry 142 Pages: 183-191
  • Peer Reviewed
• [Journal Article] Permuted tRNA genes expressed via a circular RNA intermediate in Cyanidioschyzon Merolae (2007)
  • Author(s): Soma Akiko
  • Journal Title: Science 318 Pages: 450-453
  • Peer Reviewed
• [Presentation] 枯草菌16SrRNAにおけるHelix 10領域の改変株の構築とその解析 (2010)
  • Author(s): 鈴木祥太
  • Organizer: 2010年度日本農芸化学会
  • Place of Presentation: 東京
  • Year and Date: 2010-03-30
• [Presentation] 単一rRNA保有株における増殖と胞子形成の阻害は、rRNAオペロンの重複によりサプレスされる (2010)
  • Author(s): 増田健太
  • Organizer: 2010年度日本農芸化学会
  • Place of Presentation: 東京
  • Year and Date: 2010-03-30
• [Presentation] 枯草菌新規ppGpp合成酵素YwaCによるリボソームの二量体化機構の解析 (2010)
  • Author(s): 田上和美
  • Organizer: 2010年度日本農芸化学会
  • Place of Presentation: 東京
  • Year and Date: 2010-03-30
• [Presentation] 枯草菌の正常な増殖と胞子形成に必要なrRNAオペロンのコピー数評価 (2010)
  • Author(s): 和田哲也
  • Organizer: 2010年度日本農芸化学会
  • Place of Presentation: 東京
  • Year and Date: 2010-03-29
• [Presentation] 単一rRNA保有株における増殖と胞子形成の阻害は、rRNAオペロンの重複によりサプレスされる (2010)
  • Author(s): 増田健太
  • Organizer: 2010年度日本農芸化学会大会
  • Place of Presentation: 東京、東京大学
  • Year and Date: 2010-03-29
• [Presentation] 枯草菌16SrRNAにおける Helix 10領域の改変株の構築とその解析 (2010)
  • Author(s): 鈴木将太
  • Organizer: 2010年度日本農芸化学会大会
  • Place of Presentation: 東京、東京大学
  • Year and Date: 2010-03-29
• [Presentation] 枯草菌新奇ppGpp合成酵素YwaCによるリボソームの二量体化機構の解析 (2010)
  • Author(s): 田上和美
  • Organizer: 2010年度日本農芸化学会大会
  • Place of Presentation: 東京、東京大学
  • Year and Date: 2010-03-29
• [Presentation] シアノバクテリアに存在する葉緑体特異的リボソームタンパク質PSRP-1の機能解析 (2010)
  • Author(s): 齋藤勝和
  • Organizer: 2010年度日本農芸化学会大会
  • Place of Presentation: 東京、東京大学
  • Year and Date: 2010-03-29
• [Presentation] 枯草菌の正常な増殖と胞子形成に必要なrRNAオペロンのコピー数評価 (2010)
  • Author(s): 和田哲也
  • Organizer: 2010年度日本農芸化学会大会
  • Place of Presentation: 東京、東京大学
  • Year and Date: 2010-03-28
• [Presentation] 枯草菌における増殖・胞子形成が低下したrRNAオペロン多重欠失変異株から単離された遺伝子重複を伴うサプレッサー解析 (2009)
  • Author(s): 増田健太
  • Organizer: 日本遺伝学会第81回大会
  • Place of Presentation: 松本
  • Year and Date: 2009-09-17
• [Presentation] 枯草菌における新規ppGpp合成酵素YwaCの生体内機能とリボソームの二量体化機構の解析 (2009)
  • Author(s): 田上和美
  • Organizer: 日本遺伝学会第81回大会
  • Place of Presentation: 松本
  • Year and Date: 2009-09-17
• [Presentation] 枯草菌における増殖・胞子形成能が低下したrRNAオペロン多重欠失変異株から単離された遺伝子重複を伴うサプレッサー解析 (2009)
  • Author(s): 増田健太
  • Organizer: 日本遺伝学会第81回大会
  • Place of Presentation: 松本、信州大学
  • Year and Date: 2009-09-17
• [Presentation] 枯草菌における新奇ppGpp合成酵素YwaCの生体内機能とリボソームの二量体化機構の解析 (2009)
  • Author(s): 田上和美
  • Organizer: 日本遺伝学会第81回大会
  • Place of Presentation: 松本、信州大学
  • Year and Date: 2009-09-17
• [Presentation] Isolation and characterization of suppressor mutants of th e strains carrying a single copy of rRNA operon in Bacillus subtilis. (2009)
  • Author(s): Wada Tetsuya
  • Organizer: 5th International Conference on Gram-positive Microorganisms
  • Place of Presentation: アメリカ、サンディエゴ
  • Year and Date: 2009-06-16
• [Presentation] Isolation and characterization of various rRNA gene mutants includingantibiotic resistance mutants in Bacillus subtilis. (2009)
  • Author(s): Suzuki Shota
  • Organizer: 5th International Conference on Gram-positive Microorganisms.
  • Place of Presentation: アメリカ、サンディエゴ
  • Year and Date: 2009-06-16
• [Presentation] Isolation and characterization of suppressor mutants of the strains carrying a single copy of rRNA operon in Bacillus subtilis (2009)
  • Author(s): 和田哲也
  • Organizer: 5^<th> International Conference on Gram-positive Microorganisms
  • Place of Presentation: アメリカ、サンディエゴ
  • Year and Date: 2009-06-16
• [Presentation] Isolation and characterization of various rRNA gene mutants including antibiotic resistance mutants in Bacillus subtilis (2009)
  • Author(s): 鈴木将太
  • Organizer: 5^<th> International Conference on Gram-positive Microorganisms
  • Place of Presentation: アメリカ、サンディエゴ
  • Year and Date: 2009-06-16
• [Presentation] 枯草菌における単一rRNAオペロン保有株のサプレッサーメカニズムの解析 (2009)
  • Author(s): 和田哲也
  • Organizer: 2009年度日本農芸化学会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-29
• [Presentation] 枯草菌rRNAオペロンの多コピー化に関する研究 (2009)
  • Author(s): 鈴木祥太
  • Organizer: 2009年度日本農芸化学会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-29
• [Presentation] 枯草菌における新規ppGpp合成酵素YwaCによるダイマーリボソーム形成の解析 (2009)
  • Author(s): 田上和美
  • Organizer: 2009年度日本農芸化学会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-29
• [Presentation] 枯草菌リボソームタンパク質L2に関する温度感受性変異株のサプレッサー因子yaaA遺伝子の機能解析 (2009)
  • Author(s): 谷川蔵
  • Organizer: 2009年度日本農芸化学会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-29
• [Presentation] 枯草菌における単一rRNAオペロン保有株のサプレッサーメカニズムの解析 (2009)
  • Author(s): 和田 哲也
  • Organizer: 2009年度日本農芸化学会大会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-29
• [Presentation] 枯草菌多コピー化に関する研究 (2009)
  • Author(s): 鈴木 祥太
  • Organizer: 2009年度日本農芸化学会大会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-29
• [Presentation] 枯草菌における新奇ppGpp合成遺伝子ywaCによるタイマーリボソーム形成の解析 (2009)
  • Author(s): 田上 和美
  • Organizer: 2009年度日本農芸化学会大会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-29
• [Presentation] 枯草菌リボソームタンパク質L2に関する温度感受性変異株のサプレッサー因子yaaA遺伝子の機能解析 (2009)
  • Author(s): 谷川 蔵
  • Organizer: 2009年度日本農芸化学会大会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-29
• [Presentation] Cyanidioschyzon merolae における細胞質およびミトコンドリア、葉緑体リボソームのプロテオーム解析 (2009)
  • Author(s): 佐藤 紘
  • Organizer: 2009年度日本農芸化学会大会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-29
• [Presentation] 枯草菌における新規ppGpp合成酵素YwaCの機能解析 (2008)
  • Author(s): 田上和美
  • Organizer: 日本遺伝学会第80回大会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-09-05
• [Presentation] 枯草菌単一rRNAオペロン保有株からの細胞増殖・胞子形成能の回復したサプレッサーの単離と解析 (2008)
  • Author(s): 増田健太
  • Organizer: 日本遺伝学会第80回大会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-09-05
• [Presentation] 枯草菌におけるリボソームタンパク質L2をコードするrplB遺伝子の温度感受性変異株とそのサプレッサー変異株の機能解析 (2008)
  • Author(s): 谷川蔵
  • Organizer: 日本遺伝学会第80回大会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-09-05
• [Presentation] Cyanidioschyzon merolaeにおける細胞質およびミトコンドリア、葉緑体リボソームのプロテオーム解析 (2008)
  • Author(s): 佐藤紘
  • Organizer: 日本遺伝学会第80回大会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-09-05
• [Presentation] 枯草菌における新奇ppGpp合成酵素YwaCの機能解析 (2008)
  • Author(s): 田上 和美
  • Organizer: 日本遺伝学会第80回大会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-09-05
• [Presentation] Function analysis of (p) ppGpp synthetase genes, relA, yjbA, in Bacillus subtilis. (2008)
  • Author(s): Kazumi Tagami
  • Organizer: XX International Congress of Genetics
  • Place of Presentation: ドイツ、ベルリン
  • Year and Date: 2008-07-13
• [Presentation] Studies on multiple deletion mutants of rRNA operons in Bacillus subtilis (2008)
  • Author(s): Kenta Masuda
  • Organizer: XX International Congress of Genetics
  • Place of Presentation: ドイツ、ベルリン
  • Year and Date: 2008-07-13
• [Presentation] Cyanidioschyzon merolaeにおける細胞質および葉緑体リボソームのプロテオーム解析 (2007)
  • Author(s): 佐藤 紘
  • Organizer: 日本遺伝学会第78回大会
  • Place of Presentation: 岡山
  • Year and Date: 2007-09-20
• [Presentation] 枯草菌relA,yjbM,ywaC遺伝子によるrRNAオペロンの転写制御 (2007)
  • Author(s): 村上 佳奈
  • Organizer: 日本遺伝学会第78回大会
  • Place of Presentation: 岡山
  • Year and Date: 2007-09-20
• [Presentation] 枯草菌における単-rRNAオペロン保有変異株とそのサプレッサー変異株の機能解析 (2007)
  • Author(s): 佐藤 牧子
  • Organizer: 日本遺伝学会第78回大会
  • Place of Presentation: 岡山
  • Year and Date: 2007-09-20
• [Presentation] 枯草菌におけるL2リボソームタンパク質遺伝子,rplBの温度感受性株とそのサプレッサー変異株の機能解析 (2007)
  • Author(s): 谷川 蔵
  • Organizer: 日本遺伝学会第78回大会
  • Place of Presentation: 岡山
  • Year and Date: 2007-09-20
• [Presentation] 枯草菌の16S,23SrRNA遺伝子変異による薬剤耐性株の単離 (2007)
  • Author(s): 名取 陽佑
  • Organizer: 日本遺伝学会第78回大会
  • Place of Presentation: 岡山
  • Year and Date: 2007-09-20
• [Presentation] 枯草菌における単一rRNAオペロン保有変異株とそのサプレッサー変異株の機能解析 (2007)
  • Author(s): 増田 健太
  • Organizer: 日本遺伝学会第80回大会
  • Place of Presentation: 名古屋
  • Year and Date: 2007-09-05
• [Presentation] Effect of relA and its suppressor genes on transcription activity of the rRNA operon in Bacillus subtilis (2007)
  • Author(s): Natori Yousuke
  • Organizer: 14th International Conference on Bacilli
  • Place of Presentation: イタリア、ピサ
  • Year and Date: 2007-07-27
• [Presentation] Isolation and characterization of a temperature-sensitive mutant of the rplB gene encoding the L2 ribosomal protein,and its suppressor mutant in Bacillus subtilis (2007)
  • Author(s): Osamu Tanigawa
  • Organizer: 14th International Conference on Bacilli
  • Place of Presentation: イタリア、ピサ
  • Year and Date: 2007-07-27