Analysis of tail adsorbed protein of Clostridium botulinum types C and D neurotoxin-converting phage and its host bacterial receptors

• Project/Area Number: 25460541
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bacteriology (including mycology)
• Research Institution: Kitasato University (2014-2015); University of Miyazaki (2013)
• Principal Investigator: Sakaguchi Yoshihiko 北里大学, 医学部, 助教 (70403491)
• Co-Investigator(Kenkyū-buntansha): MATSUZAKI Shigenobu 高知大学, 医歯学系, 准教授 (00190439) ; UCHIYAMA Jumpei 麻布大学, 獣医学部, 講師 (20574619)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: ボツリヌス菌 / 人畜共通感染症 / ボツリヌス毒素遺伝子 / バクテリオファージ / 水平伝播 / ファージ変換 / ゲノム解析 / プロテオーム解析

Outline of Final Research Achievements

Because Clostridium botulinum types C and D produce botulinum neurotoxin, causing serious symptom botulism, they are recognized as severe pathogen you should prevent spreading in food and animal industries. The botulinum neurotoxin genes are coded on the prophage genomes, and so inhibition of lateral gene transfer mediated by bacteriophage (phage) can be one of the preventions for the disease. Thus, we investigated the adsorption mechanism of C. botulinum type C neurotoxin-converting phage, phage c-st on the genetic point. Based on the comparative genomics and identification of phage virion genes, we got at a speculation that several genes associated with phage adsorption. We are in advanced studies of the findings.

Research Products

• [Journal Article] Characterization of Pseudomonas aeruginosa phage KPP21 belonging to family Podoviridae genus N4-like viruses isolated in Japan (2016)
  • Author(s): Shigehisa, R., Uchiyama, J., Kato S, Takemura-Uchiyama, I, Yamaguchi, K., Miyata, R., Ujihara, T., Sakaguchi, Y., Okamoto, N., Shimakura, H., Daibata, M., Sakaguchi, M., and Matsuzaki, S.
  • Journal Title: Microbiology and Immunology Volume: 60 Issue: 1 Pages: 64-67
  • DOI: 10.1111/1348-0421.12347
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Genomics of Clostridium botulinum group III strains (2015)
  • Author(s): Sakaguchi, Y., Suzuki, T., Yamamoto, Y., Nishikawa, A., Oguma, K.
  • Journal Title: Res. Microbiol. Volume: 166 Issue: 4 Pages: 318-325
  • DOI: 10.1016/j.resmic.2014.07.016
  • Peer Reviewed / Open Access
• [Journal Article] 「シリーズ！！ボツリヌス菌による疾患（ボツリヌス症）」第一回 発見の歴史と疫学 (2015)
  • Author(s): 阪口義彦
  • Journal Title: サラヤ情報誌「衛生の友」 Volume: －
• [Journal Article] 「シリーズ！！ボツリヌス菌による疾患（ボツリヌス症）」第二回 病原因子と作用機序 (2015)
  • Author(s): 阪口義彦
  • Journal Title: サラヤ情報誌「衛生の友」 Volume: －
• [Journal Article] 「シリーズ！！ボツリヌス菌による疾患（ボツリヌス症）」第三回 臨床症状（ボツリヌス食中毒、乳児ボツリヌス症、創傷ボツリヌス症） (2015)
  • Author(s): 阪口義彦
  • Journal Title: サラヤ情報誌「衛生の友」 Volume: －
• [Journal Article] Complete genome sequence of a Pseudomonas aeruginosa bacteriophage KPP23 belonging to family Siphoviridae (2014)
  • Author(s): Yamaguchi K, Miyata R, Shigehisa R, Uchiyama J, Takemura-Uchiyama I, Kato S, Ujihara T, Sakaguchi Y, Daibata M, Matsuzaki S
  • Journal Title: Genome Announcements Volume: 2(3) Issue: 3
  • DOI: 10.1128/genomea.00233-14
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of a novel Pseudomonas aeruginosa bacteriophage, KPP25, of the family Podoviridae (2014)
  • Author(s): Miyata R, Yamaguchi K, Uchiyama J, Shigehisa R, Takemura-Uchiyama I, Kato S, Ujihara T, Sakaguchi Y, Daibata M, Matsuzaki S
  • Journal Title: Virus Research Volume: 189 Pages: 43-46
  • DOI: 10.1016/j.virusres.2014.04.019
  • Peer Reviewed / Open Access
• [Journal Article] DNAX-activating protein 10 (DAP10) membrane adaptor associates with receptor for advanced glycation end products (RAGE) and modulates the RAGE-triggered signaling pathway in human keratinocytes (2014)
  • Author(s): Sakaguchi M, Murata H, Aoyama Y, Hibino T, Putranto EW, Ruma IM, Inoue Y, Sakaguchi Y, Yamamoto K, Kinoshita R, Futami J, Kataoka K, Iwatsuki K, Huh NH
  • Journal Title: J Biol Chem Volume: 289 Issue: 34 Pages: 23389-23402
  • DOI: 10.1074/jbc.m114.573071
  • NAID: 120006333870
  • Peer Reviewed / Open Access
• [Journal Article] Complete nucleotide sequence of a plasmid containing the botulinum neurotoxin gene in Clostridium botulinum type B strain 111 isolated from an infant patient in Japan. (2014)
  • Author(s): Hosomi, K., Sakaguchi, Y., Kohda, T., Gotoh, K., Motooka, D., Nakamura, S., Umeda, K., Iida, T., Kozaki, S., Mukamoto, M.
  • Journal Title: Mol. Genet. Genomics Volume: 289(6) Issue: 6 Pages: 1267-1274
  • DOI: 10.1007/s00438-014-0887-4
  • Peer Reviewed / Open Access
• [Journal Article] Draft Genome Sequence of Clostridium botulinum Type B Strain Osaka05, Isolated from an Infant Patient with Botulism in Japan. (2014)
  • Author(s): Sakaguchi, Y., Hosomi, K., Uchiyama, J., Ogura, Y., Umeda, K., Sakaguchi, M., Kohda, T., Mukamoto, M., Misawa, N., Matsuzaki, S., Hayashi, T., Kozaki, S.
  • Journal Title: Genome Announc. Volume: 23 Issue: 1
  • DOI: 10.1128/genomea.01010-13
  • Peer Reviewed / Open Access
• [Journal Article] Intragenus generalized transduction in Staphylococcus spp. by a novel giant phage. (2014)
  • Author(s): Uchiyama J, Takemura-Uchiyama I, Sakaguchi Y, Gamoh K, Kato SI, Daibata M, Ujihara T, Misawa N, Matsuzaki S.
  • Journal Title: ISME J. Volume: in press Issue: 9 Pages: 1949-1952
  • DOI: 10.1038/ismej.2014.29
  • Peer Reviewed / Open Access
• [Journal Article] Intracellular trafficking of Clostridium botulinum C2 toxin (2014)
  • Author(s): Nagahama, M., Takahashi, C., Aoyanagi, K., Tashiro, R., Kobayashi, K., Sakaguchi, Y., Ishidoh, K., Sakurai, J.
  • Journal Title: Toxicon Volume: 82 Pages: 76-82
  • DOI: 10.1016/j.toxicon.2014.02.009
  • Peer Reviewed
• [Presentation] The genome sequence of Clostridium botulinum type C phage and functional analysis of gene products (2016)
  • Author(s): Sakaguchi, Y., Uchiyama, J., Ogura, Y., Yamamoto, Y., Matsuzaki, S., Hayashi, T., Matsushita, O., Oguma, K., Hayashi, S.
  • Organizer: 第89回日本細菌学会総会
  • Place of Presentation: 大阪国際交流センター（大阪府大阪市）
  • Year and Date: 2016-03-23
• [Presentation] 緑膿菌PAO1株とファージKPP22の短期間進化的軍拡競走の解析 (2016)
  • Author(s): 那須川忠弥、内山淳平、鈴木仁人、宮田玲奈、山口琴絵、内山伊代、阪口義彦、柴山恵吾、阪口雅弘、松崎茂展
  • Organizer: 第89回日本細菌学会総会
  • Place of Presentation: 大阪国際交流センター（大阪府大阪市）
  • Year and Date: 2016-03-23
• [Presentation] 早期胃癌ESD後再発予測におけるH. pylori－J抗原を用いたIgG サブクラスの有用性 (2016)
  • Author(s): 横田憲治、松下 治、山本由弥子、美間健彦、後藤和義、林 俊治、阪口義彦
  • Organizer: 第89回日本細菌学会総会
  • Place of Presentation: 大阪国際交流センター（大阪府大阪市）
  • Year and Date: 2016-03-23
• [Presentation] C型ボツリヌス毒素変換ファージのゲノム比較と遺伝子機能解析 (2015)
  • Author(s): 阪口義彦、内山淳平、Oliva, M. A.、Martin-Galiano, A. J.、Andreu, J. M.、小椋義俊、山本 由弥子、松崎茂展、林 哲也、小熊惠二、林 俊治
  • Organizer: 第98回日本細菌学会関東支部総会
  • Place of Presentation: 東京歯科大学（東京都）
  • Year and Date: 2015-10-29
• [Presentation] C型ボツリヌス毒素変換ファージの生存に必要なゲノム領域の解析 (2015)
  • Author(s): 阪口義彦、内山淳平、 Oliva, M. A.、Martin-Galiano, A. J.、Andreu, J. M.、小椋義俊、山本 由弥子、松崎茂展、林 哲也、小熊惠二、林 俊治
  • Organizer: 平成27年度AKPS研究集会、第18回北里微生物アカデミー
  • Place of Presentation: 北里大学（神奈川県相模原市）
  • Year and Date: 2015-08-06
• [Presentation] ピロリ菌ファージの分離と解析 (2015)
  • Author(s): 内山淳平、竹内 啓晃、加藤 伸一郎、阪口義彦、蒲生啓司、氏原隆子、内山伊代、島倉秀勝、今西市朗、阪口雅弘、松崎茂展
  • Organizer: 第2回ファージセラピー臨床応用検討会
  • Place of Presentation: 東京工業大学（神奈川県横浜市）
  • Year and Date: 2015-07-31
• [Presentation] Analysis of botulinum neurotoxin type B6 gene-encoding plasmid in Clostridium botulinum type B (2015)
  • Author(s): Sakaguchi, Y., Uchiyama, J., Hosomi, K., Kohda, T., Matsuzaki, S., Ogura, Y., Hayashi, T., Hayashi, S., Kozaki, S., Mukamoto, M.
  • Organizer: 第88回日本細菌学会総会
  • Place of Presentation: 長良川国際会議場（岐阜県岐阜市）
  • Year and Date: 2015-03-26 – 2015-03-28
• [Presentation] 各種ファージに対する黄色ブドウ球菌受容体の解析 (2015)
  • Author(s): 内山淳平、黒川健児、阪口義彦、内山（竹村）伊代、氏原隆子、村上雅尚、大畑雅典、松崎茂展
  • Organizer: 第88回日本細菌学会総会
  • Place of Presentation: 長良川国際会議場（岐阜県岐阜市）
  • Year and Date: 2015-03-26 – 2015-03-28
• [Presentation] 緑膿菌感染症に対するファージ療法と化学療法の併用療法の可能性検討 (2015)
  • Author(s): 重久 立、内山淳平、宮田怜奈、山口琴絵、内山伊代、阪口義彦、氏原隆子、大畑雅典、松崎茂展
  • Organizer: 第88回日本細菌学会総会
  • Place of Presentation: 長良川国際会議場（岐阜県岐阜市）
  • Year and Date: 2015-03-26 – 2015-03-28
• [Presentation] ピロリ菌におけるKHP30様ファージの溶原性に関する研究 (2014)
  • Author(s): 内山淳平、内山（竹村）伊代、阪口義彦、竹内啓晃、村上政尚、氏原隆子、大畑雅典、松崎茂展
  • Organizer: 第67回日本細菌学会中国・四国支部総会
  • Place of Presentation: 徳島文理大学薬学部（徳島県徳島市）
  • Year and Date: 2014-10-04 – 2014-10-05
• [Presentation] Generalized transduction experiment in vitro among Staphylococcus spp., using a novel giant phage S6. (2014)
  • Author(s): Uchiyama, J., Takemura-Uchiyama, I., Sakaguchi, Y., Kato, S., Gamoh, K., Misawa, N., Murakami, M., Daibata, M., Matsuzaki, S.
  • Organizer: 第5回ファージ研究会
  • Place of Presentation: 三重大学生物資源学部（三重県津市）
  • Year and Date: 2014-09-04 – 2014-09-05
• [Presentation] Analysis of botulinum neurotoxin type G gene-encoding plasmid in Clostridium argentinense strain 2740. (2014)
  • Author(s): Sakaguchi, Y., Uchiyama, J., Suzuki, T., Yamamoto, Y., Hosomi, K., Kohda, T., Matsuzaki, S., Hayashi, T., Mukamoto, M., Oguma, K.
  • Organizer: International Union of Microbiological Societies (IUMS)
  • Place of Presentation: Montreal Convention Center (Montreal, Canada)
  • Year and Date: 2014-07-27 – 2014-08-01
• [Presentation] Generalized transduction in vitro among Staphylococcus spp., using a novel phage S6. (2014)
  • Author(s): Uchiyama, J., Takemura-Uchiyama, I., Sakaguchi, Y., Kato, S., Gamoh, K., Misawa, N., Daibata, M., Matsuzaki, S.
  • Organizer: International Union of Microbiological Societies (IUMS)
  • Place of Presentation: Montreal Convention Center (Montreal, Canada)
  • Year and Date: 2014-07-27 – 2014-08-01
• [Presentation] KHP30-like phages among clinical isolates of Helicobacter pylori. (2014)
  • Author(s): Takemura-Uchiyama, I., Uchiyama, J., Takeuchi, H., Gamoh, K., Sakaguchi, Y., Kato, S., Ujihara, T., Daibata, M., Matsuzaki, S.
  • Organizer: International Union of Microbiological Societies (IUMS)
  • Place of Presentation: Montreal Convention Center (Montreal, Canada)
  • Year and Date: 2014-07-27 – 2014-08-01
• [Presentation] Examination of generalized transduction among Staphylococcus spp. using a novel phage S6 (2014)
  • Author(s): Uchiyama, J., Takemura-Uchiyama, I., Sakaguchi, Y., Gamoh, K., Misawa, N., Daibata, M., Matsuzaki, S.
  • Organizer: The 13th KMS Research meeting
  • Place of Presentation: Kochi, Japan
• [Presentation] Phage therapy experiment against staphylococcal lung-derived septic mouse model (2014)
  • Author(s): Takemura-Uchiyama, I., Uchiyama, J., Sakaguchi, Y. Ujihara, T., Daibata, M., Matsuzaki, S.
  • Organizer: The 13th KMS Research meeting
  • Place of Presentation: Kochi, Japan
• [Presentation] Estimated distribution of KHP30-like bacteriophages in the stomach, using clinical Helicobacter pylori isolates (2014)
  • Author(s): Uchiyama, J., Takeuchi, H., Sakaguchi, Y., Takemura-Uchiyama, I., Kato, S., Gamoh, K., Ujihara, T., Daibata, M., Matsuzaki, S.
  • Organizer: The 12th Japan-Korea International Symposium on Microbiology (XII-JKISM)
  • Place of Presentation: Tokyo, Japan
• [Presentation] Analysis of botulinum neurotoxin type G gene-encoding plasmid in Clostridium argentinense (2014)
  • Author(s): Sakaguchi, Y., Uchiyama, J., Suzuki, T., Yamamoto, Y., Ogura, Y., Hosomi, K., Matsuzaki, S., Hayashi, T., Kozaki, S., Oguma, K.
  • Organizer: The 87th Annual Meeting of Japanese Society for Bacteriology
  • Place of Presentation: Tokyo, Japan
• [Presentation] Examination of generalized transduction among Staphylococcus spp., using a novel phage S6 (2014)
  • Author(s): Uchiyama, J., Takemura-Uchiyama, I., Sakaguchi, Y., Misawa, N., Daibata, M., Matsuzaki, S.
  • Organizer: The 87th Annual Meeting of Japanese Society for Bacteriology
  • Place of Presentation: Tokyo, Japan
• [Presentation] Estimation of Helicobacter pylori phage ecology (2014)
  • Author(s): Matsuzaki, S., Uchiyama, J., Takeuchi, H., Takemura-Uchiyama, I., Sakaguchi, Y., Ujihara, T., Daibata, M.
  • Organizer: The 87th Annual Meeting of Japanese Society for Bacteriology
  • Place of Presentation: Tokyo, Japan
• [Presentation] Phage therapy experiment against staphylococcal lung-derived septic mouse model (2014)
  • Author(s): Takemura-Uchiyama, I., Uchiyama, J., Sakaguchi, Y., Ujihara, T., Daibata, M., Matsuzaki, S.
  • Organizer: The 87th Annual Meeting of Japanese Society for Bacteriology
  • Place of Presentation: Tokyo, Japan
• [Presentation] Bacterial detection system for Staphylococcus aureus based on bacteriophage tail adsorption protein (2013)
  • Author(s): Uchiyama, J., Uchiyama-Takemura, I., Watanabe, S., Sakaguchi, Y., Daibata, M., Matsuzaki, S.
  • Organizer: The 28th International Congress of Chemotherapy and Infection (ICC), Incorporating the 14th Asia-Pacific Congress of Clinical Microbiology and Infection
  • Place of Presentation: Yokohama, Japan
• [Presentation] Analysis of botulinum neurotoxin-encoding plasmid in of Clostridium argentinense (2013)
  • Author(s): Sakaguchi, Y., Uchiyama, J., Suzuki, T., Yamamoto, Y., Ogura, Y., Sakaguchi, M., Tsuda, C., Matsuzaki, S., Hayashi, T., Oguma, K.
  • Organizer: FEMS2013, The 5th Congress of European Microbiologists
  • Place of Presentation: Leipzig, Germany
• [Presentation] Phage therapy experiment for staphylococcal lung infection (2013)
  • Author(s): Uchiyama, J., Sakaguchi, Y., Takemura-Uchiyama, I., Kato, S., Ujihara, T., Daibata, M., Matsuzaki, S.
  • Organizer: FEMS2013, The 5th Congress of European Microbiologists
  • Place of Presentation: Leipzig, Germany
• [Presentation] A novel type of spherical bacteriophage, Helicobacter pylori bacteriophage KHP30 (2013)
  • Author(s): Uchiyama, J., Takeuchi, H., Sakaguchi, Y., Gamo, K, Kato, S., Takemura-Uchiyama, I., Ujihara, T., Daibata, M., Matsuzaki, S.
  • Organizer: FEMS2013, The 5th Congress of European Microbiologists
  • Place of Presentation: Leipzig, Germany
• [Presentation] 新奇ファージS6の分離とブドウ球菌における普遍形質導入の検討 (2013)
  • Author(s): 内山淳平、内山（竹村）伊代、蒲生恵司、阪口義彦、三澤尚明、大畑雅典、松崎茂展
  • Organizer: 第58回日本ブドウ球菌研究会
  • Place of Presentation: 東京
• [Presentation] 黄色ブドウ球菌性肺感染症に対するファージ療法の可能性の検討 (2013)
  • Author(s): 松崎茂展、内山淳平、内山（竹村）伊代、阪口義彦、加藤 伸一郎、氏原隆子、大畑雅典
  • Organizer: 第58回日本ブドウ球菌研究会
  • Place of Presentation: 東京
• [Presentation] 新規ファージ S6 によるブドウ球菌間の普遍形質導入の検討 (2013)
  • Author(s): 内山淳平、内山(竹村)伊代、阪口義彦、三澤尚明、大畑雅典、松崎茂展
  • Organizer: 第66回日本細菌学会中国・四国支部総会
  • Place of Presentation: 広島
• [Presentation] B型ボツリヌス菌が保有するプラスミドの芽胞形成への関与
  • Author(s): 細見晃司、阪口義彦、後藤和義、幸田知子、飯田哲也、小崎俊司、向本雅郁
  • Organizer: 第66回日本細菌学会関西支部総会
  • Place of Presentation: 大阪
• [Book] 化学療法の領域、III嫌気性菌、破傷風とボツリヌス症 (2013)
  • Author(s): 小熊惠二、阪口義彦、杉本 央、山本 由弥子
  • Publisher: 医薬ジャーナル社