Novel extracellular function of molecular chaperones and mechanisms of biofilm formation in pathogenic bacteria

• Project/Area Number: 24780079
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Applied microbiology
• Research Institution: Jikei University School of Medicine
• Principal Investigator: SUGIMOTO SHINYA 東京慈恵会医科大学, 医学部, 講師 (60464393)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2013: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2012: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: バイオフィルム / 分子シャペロン / ClpB / DnaK / マトリクス / 黄色ブドウ球菌 / 抗体 / 阻害剤 / Myricetin / persister / アミロイド線維 / curli / 細胞外マトリクス / 病原細菌 / 線虫 / ブドウ球菌

Outline of Final Research Achievements

Molecular chaperones play key roles in protein quality control in the cell. We found that cytoplasmic molecular chaperones ClpB and DnaK are incorporated into extracellular matrixes of Staphylococcus aureus and are involved in biofilm development. We here report that extracellularly excreted ClpB promotes biofilm formation in an oligomerization- and disaggregation activity-independent manner. Inhibition of these molecular chaperones using antibodies or small compounds leads to prevention of biofilms and persisters. These findings provide a novel mechanistic insight into molecular chaperones as well as a therapeutic strategy for chronic infectious diseases.

Research Products

• [Journal Article] A refined technique for extraction of extracellular matrixes from bacterial biofilms and its applicability (2015)
  • Author(s): Akio Chiba, Shinya Sugimoto, Fumiya Sato, Seiji Hori, Yoshimitsu Mizunoe
  • Journal Title: Microbial Biotechnology Volume: 8 Pages: 392-403
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A refined technique for extraction of extracellular matrices from bacterial biofilms and its applicability (2015)
  • Author(s): Akio Chiba, Shinya Sugimoto, Fumiya Sato, Seiji Hori, Yoshimitsu Mizunoe
  • Journal Title: Microbial Biotechnology Volume: - Issue: 3 Pages: 392-403
  • DOI: 10.1111/1751-7915.12155
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Novel strategy for biofilm inhibition by using small molecules targeting molecular chaperone DnaK (2015)
  • Author(s): Ken-ichi Arita-Morioka, Kunitoshi Yamanaka, Yoshimitsu Mizunoe, Teru Ogura, Shinya Sugimoto
  • Journal Title: Antimicrobial Agents and Chemotherapy Volume: 59(1) Issue: 1 Pages: 633-641
  • DOI: 10.1128/aac.04465-14
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Thioflavin T as a fluorescence probe for monitoring RNA metabolism at molecular and cellular levels (2015)
  • Author(s): Shinya Sugimoto, Ken-ichi Arita-Morioka, Yoshimistu Mizunoe, Kunitoshi Yamanaka, Teru Ogura
  • Journal Title: Nucleic Acids Research Volume: -
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Immuno-Electron Microscopy of Primary Cell Cultures from GeneticallyModified Animals in Liquid by Atmospheric Scanning Electron Microscopy (ASEM) (2014)
  • Author(s): T.Kinoshita, Y.Mori, K.Hirano, S.Sugimoto, K.Okuda, S.Matsumoto, T.Namiki, T.Ebihara, M.Kawata, H.Nishiyama, M.Sato, M.Suga, K.Higashiyama, K.Sonomoto, Y.Mizunoe, S.Nishihara, C.Sato
  • Journal Title: Microscopy and Microanalysis Volume: 20 Issue: 2 Pages: 469-483
  • DOI: 10.1017/s1431927614000178
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Effects of bacteriocins on methicillin-resistant Staphy lococcus a ureus biofilm. Antimicrob (2013)
  • Author(s): Ken-ichi Okuda, T akeshi Zendo, Shinya Sugimoto, T adayuki Iwase, Akiko T ajima, Satomi Y amada, Kenji Sonomoto & Y oshimitsu Mizunoe
  • Journal Title: Agents Chemother Volume: 57 Issue: 11 Pages: 5572-5579
  • DOI: 10.1128/aac.00888-13
  • Peer Reviewed
• [Journal Article] A simple assay for measuring catalase activity: a visual approach. (2013)
  • Author(s): Iwase T, Tajima A, Sugimoto S, Okuda K, Hironaka I, Kamata Y, Takada K, Mizunoe Y.
  • Journal Title: Sci Rep Volume: 3 Issue: 1 Pages: 3081-3081
  • DOI: 10.1038/srep03081
  • Peer Reviewed
• [Journal Article] Staphylococcus epidermidis Esp degrades specific proteins associated with Staphylococcus aureus biofilm formation and host-pathogen interaction. (2013)
  • Author(s): Sugimoto S, Iwamoto T, Takada K, Okuda K, Tajima A, Iwase T, Mizunoe Y.
  • Journal Title: J Bacteriol Volume: 195 Issue: 8 Pages: 1645-1655
  • DOI: 10.1128/jb.01672-12
  • Peer Reviewed
• [Journal Article] Glucose triggers ATP secretion from bacteria in a growth-phase-dependent manner. (2013)
  • Author(s): Hironaka I, Iwase T, Sugimoto S, Okuda K, Tajima A, Yanaga K, Mizunoe Y.
  • Journal Title: Appl Environ Microbiol Volume: 79 Issue: 7 Pages: 2328-35
  • DOI: 10.1128/aem.03871-12
  • Peer Reviewed
• [Presentation] 細胞外アミロイド産生におけるコシャペロン非依存的な分子シャペロンDnaKの機能 (2015)
  • Author(s): ○杉本真也，有田-森岡健一，山中邦俊，小椋光，水之江義充
  • Organizer: 第38回日本分子生物学会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] Inhibition of bacterial biofilms by small compounds targeting molecular chaperone DnaK (2015)
  • Author(s): Ken-ichi Arita-Morioka, Kunitoshi Yamanaka, Yoshimistu Mizunoe, Teru Ogura, ○Shinya Sugimoto
  • Organizer: Microbial Stress: From Molecule to Systems
  • Place of Presentation: Sitges
  • Year and Date: 2015-11-12
  • Int'l Joint Research
• [Presentation] 8型分泌装置の発動における細胞質分子シャペロンの機能 (2015)
  • Author(s): 〇杉本真也
  • Organizer: 第98回日本細菌学会関東支部総会
  • Place of Presentation: 東京
  • Year and Date: 2015-10-29
  • Invited
• [Presentation] DnaK/Hsp70シャペロンシステムはVIII型分泌装置の発現と品質を制御する (2015)
  • Author(s): ○杉本真也，有田健一，山中邦俊，小椋光，水之江義充
  • Organizer: 第12回21世紀大腸菌研究会
  • Place of Presentation: 滋賀
  • Year and Date: 2015-06-04
• [Presentation] 分子シャペロンDnaKをターゲットにした小分子化合物を用いた新規バイオフィルム阻害法の開発 (2015)
  • Author(s): ○有田健一，山中邦俊，水之江義充，小椋光，杉本真也
  • Organizer: 第12回21世紀大腸菌研究会
  • Place of Presentation: 滋賀
  • Year and Date: 2015-06-04
• [Presentation] DnaK/Hsp70 シャペロンシステムは8 型分泌装置の発現と品質を制御する (2015)
  • Author(s): 杉本 真也, 有田（森岡） 健一, 山中 邦俊, 小椋 光, 水之江 義充
  • Organizer: 第88回日本細菌学会総会
  • Place of Presentation: 岐阜
  • Year and Date: 2015-03-26 – 2015-03-28
• [Presentation] Extracellular small RNA はバイオフィルムの構造維持に重要である (2015)
  • Author(s): 千葉 明生, 杉本 真也, 水之江 義充
  • Organizer: 第88回日本細菌学会総会
  • Place of Presentation: 岐阜
  • Year and Date: 2015-03-26 – 2015-03-28
• [Presentation] 分子シャペロンDnaKはタンパク質のフォールディングと局在化を介してバイオフィルム形成を制御する (2014)
  • Author(s): 杉本 真也, 有田（森岡） 健一, 山中 邦俊, 小椋 光, 水之江 義充
  • Organizer: 第37回日本分子生物学会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 分子シャペロンDnaKを標的とする低分子化合物を用いた新たなバイオフィルム阻害法の開発 (2014)
  • Author(s): 有田(森岡) 健一, 山中 邦俊, 水之江 義充, 小椋 光, 杉本 真也
  • Organizer: 第37回日本分子生物学会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 迅速な非侵襲的バイオフィルムマトリクス抽出法の開発 (2014)
  • Author(s): 千葉 明生, 杉本 真也, 佐藤 文哉, 堀 誠治, 水之江 義充
  • Organizer: 第37回日本分子生物学会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Fine structures of biofilms in solution visualized by atmospheric scanning electron microscopy (2014)
  • Author(s): Shinya Sugimoto, Ken-ichi Okuda, Mari Sato, Chikara Sato, Yoshimitsu Mizunoe
  • Organizer: 19th World Congress on Advances in Oncology and 17th World Congress on Advances in Oncology
  • Place of Presentation: アテネ
  • Year and Date: 2014-10-07 – 2014-10-13
  • Invited
• [Presentation] DnaK阻害剤を用いた慢性細菌感染症の制御戦略 (2014)
  • Author(s): 有田 (森岡) 健一, 山中 邦俊, 水之江 義充, 小椋 光, 杉本 真也
  • Organizer: 第66回日本生物工学会大会
  • Place of Presentation: 札幌
  • Year and Date: 2014-09-09 – 2014-09-11
• [Presentation] 迅速な非侵襲的バイオフィルムマトリクス抽出法の開発． (2014)
  • Author(s): ○千葉明生、佐藤文哉、水之江義充、杉本真也
  • Organizer: 第87回日本細菌学会総会
  • Place of Presentation: 東京
• [Presentation] Effects of bacteriocins on methicillin-resistant Staphylococcus aureus biofilm. (2014)
  • Author(s): ○奥田賢一，杉本真也，岩瀬忠行，田嶌亜紀子，水之江義充
  • Organizer: 第87回日本細菌学会総会
  • Place of Presentation: 東京
• [Presentation] Staphylococcus epidermidis Esp degrades specific proteins associated with Staphylococcus aureus biofilm formation and host-pathogen interaction. (2013)
  • Author(s): 〇Sugimoto S., Iwamoto T., Takada K., Okuda K., Tajima A, Iwase T, Chiba A., Mizunoe Y.
  • Organizer: 5th FEMS Congress 2013
  • Place of Presentation: Leipzig
• [Presentation] Excreted AAA+ chaperone ClpB stimulates bacterial biofilm formation. (2013)
  • Author(s): 〇Sugimoto S., Mizunoe Y.
  • Organizer: 10th International Conference on AAA+ Proteins (EMBO Workshop on AAA+ proteins: From mechanisms and disease to targets).
  • Place of Presentation: Neuss
• [Presentation] 大気圧走査電子顕微鏡によるバクテリアの多細胞的形態“バイオフィルム”の液中高分解能観察． (2013)
  • Author(s): 〇杉本真也、奥田賢一、千葉明生、佐藤主税、水之江義充
  • Organizer: 第36回日本分子生物学会
  • Place of Presentation: 神戸
• [Presentation] 大気圧走査電子顕微鏡による水中での細胞の観察：乳酸菌と細胞外構造． (2013)
  • Author(s): 杉本真也、奥田賢一、並木健、東山堅一、佐藤真理、西山英利、須賀三雄、海老原達彦、松本俊介、園元謙二、水之江義充、〇佐藤主税
  • Organizer: 第36回日本分子生物学会
  • Place of Presentation: 神戸
• [Presentation] Bacterial ATP secretion (2013)
  • Author(s): Hironaka I, Iwase T, Sugimoto S, Okuda K, Tajima A, Yanaga K, Mizunoe Y
  • Organizer: Annual Conference of the Association for General and Applied Microbiology.
  • Place of Presentation: Bremen, Germany
• [Presentation] 細菌によるATP分泌機構の解析 (2013)
  • Author(s): 弘中一平，岩瀬忠行，杉本真也，奥田賢一，田嶌亜紀子，水之江義充
  • Organizer: 第86回日本細菌学会総会
  • Place of Presentation: 千葉
• [Presentation] Quality control of staphylococcal biofilms by excreted molecular chaperones DnaK and ClpB. (2012)
  • Author(s): Sugimoto S, Okuda K, Tajima A, Iwase T, Mizunoe Y.
  • Organizer: EMBO/EMBL Symposium, Quality Control -From Molecules to Organelles-
  • Place of Presentation: Heidelberg, Germany
• [Presentation] MRSA のバイオフィルムマトリクスタンパク質の網羅的解析と時空間的動態 (2012)
  • Author(s): 杉本真也、奥田賢一、佐藤主税、水之江義充
  • Organizer: 第95回日本細菌学会関東支部総会
  • Place of Presentation: 東京
• [Presentation] 細胞外に排出された細胞質分子シャペロンによるバイオフィルム形成制御メカニズム (2012)
  • Author(s): 杉本真也、千葉明夫、弘中一平、奥田賢一、田嶌亜紀子、岩瀬忠行、水之江義充
  • Organizer: 第35回日本分子生物学会
  • Place of Presentation: 福岡
• [Presentation] メチシリン耐性黄色ブドウ球菌バイオフィルムに対するバクテリオシンの殺菌効果 (2012)
  • Author(s): 奥田賢一、善藤 威史、田嶌亜紀子、岩瀬忠行、杉本真也、弘中一平、千葉明生、園元謙二、水之江義充
  • Organizer: 第35回日本分子生物学会
  • Place of Presentation: 福岡
• [Presentation] バクテリアの多細胞的形態“バイオフィルム”の液中高分解能観察．
  • Author(s): 〇杉本真也
  • Organizer: 山形大学理学部・テニュアトラック合同シンポジウム
  • Place of Presentation: 山形
  • Invited
• [Book] 化学療法の領域：バイオフィルム細胞外マトリクスの分離・解析 (2015)
  • Author(s): 水之江義充，千葉明生，岩瀬忠行，杉本真也
  • Total Pages: 8
  • Publisher: 医学ジャーナル社
• [Remarks] 東京慈恵会医科大学細菌学講座
  • URL: http://square.umin.ac.jp/saikin/
• [Remarks] 細菌学講座 東京慈恵会医科大学 基礎・臨床講座
  • URL: http://www.jikei.ac.jp/academic/course/11_saikin.html
• [Remarks] 東京慈恵会医科大学医学部細菌学講座ホームページ
  • URL: http://square.umin.ac.jp/saikin/
• [Remarks] 東京慈恵会医科大学細菌学講座HP
  • URL: http://square.umin.ac.jp/saikin/
• [Remarks] 細菌学講座/東京慈恵会医科大学
  • URL: http://www.jikei.ac.jp/academic/course/11_saikin.html