Elucidation of the mechanism of outer membrane vesicles production in Gram-negative bacteria

• Project/Area Number: 26860293
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Bacteriology (including mycology)
• Research Institution: Kyoto University (2015-2016); University of Miyazaki (2014)
• Principal Investigator: Kazunori Murase 京都大学, 医学研究科, 研究員 (40710869)
• Research Collaborator: SHIRAISHI Tsukasa 札幌医科大学医学部, 微生物学講座, 助教
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 外膜小胞 / 国際共同研究 / HlyF / グラム陰性細菌 / 国際情報交換（フランス）

Outline of Final Research Achievements

The hemolysin F (hlyF) gene is epidemiologically associated with virulent strains of avian pathogenic E. coli and human neonatal meningitis-associated E. coli. We demonstrated that culture supernatants of E. coli expressing HlyF induced autophagy in eukaryotic cells. This phenotype coincided with an enhanced production of outer membrane vesicles (OMVs) by bacteria expressing HlyF. The HlyF protein displays a predicted catalytic domain of the short chain dehydrogenase/reductase superfamily. This conserved domain was involved the ability of HlyF to promote the production of OMVs.
This study is the first evidence that pathogenic bacteria produce a virulence factor directly involved in the production of OMVs and expected that HlyF-dependent OMV production could be one of the method for the development of vaccine.

Research Products

• [Int'l Joint Research] フランス国立保健医学研究所（INSERM）/フランス国立農学研究所（INRA）/フランス国立科学研究センター（CNRS）(フランス)
• [Int'l Joint Research] フランス国立保健医学研究機構（INSERM）/フランス国立農学研究所（INRA）/フランス国立科学研究センター（CNRS）(フランス)
• [Int'l Joint Research] アルマンフラピエ研究所（INRS Armand-Frappier）(カナダ)
• [Journal Article] Distinct interacting core taxa in co-occurrence networks enable discrimination of polymicrobial oral diseases with similar symptoms (2016)
  • Author(s): Shiba T, Watanabe T, Kachi H, Koyanagi T, Maruyama N, Murase K, Takeuchi Y, Maruyama F, Izumi Y, Nakagawa I
  • Journal Title: Scientific reports Volume: 6 Issue: 1 Pages: 30997-30997
  • DOI: 10.1038/srep30997
  • NAID: 120005841407
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] DNA-based culture-independent analysis detects the presence of Group A Streptococcus in throat samples from healthy adults in Japan (2016)
  • Author(s): T. Kulkarni, F. Maruyama, C. Aikawa, T. Nozawa, K. Murase, I. Nakagawa.
  • Journal Title: BMC Microbiology Volume: 16(1) Issue: 1 Pages: 237-237
  • DOI: 10.1186/s12866-016-0858-5
  • NAID: 120006223635
  • Peer Reviewed / Open Access
• [Journal Article] Hlyf Produced By Extraintestinal Pathogenic Escherichia Coli Is A Virulence Factor That Regulates Outer Membrane Vesicle Biogenesis (2015)
  • Author(s): Kazunori Murase, Patricia Martin, Gaëlle Porcheron, Sébastien Houle, Emmanuelle Helloin, Marie Pénary, Jean-Philippe Nougayrède, Charles M. Dozois, Tetsuya Hayashi, and Eric Oswald
  • Journal Title: Journal of Infectious Diseases Volume: 213(5) Issue: 5 Pages: 856-865
  • DOI: 10.1093/infdis/jiv506
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] New mechanisms of outer membrane vesicles in Gram-negative bacteria (2016)
  • Author(s): Kazunori Murase
  • Organizer: The International Seminar on Applied Microbial Ecology
  • Place of Presentation: Temuco, Chile
  • Year and Date: 2016-11-25
  • Int'l Joint Research / Invited
• [Presentation] HlyFによって誘導される外膜小胞の産生機構の解明 (2016)
  • Author(s): 村瀬一典, Patricia Martin, Jean-Philippe Nougayrede, 林哲也, Eric Oswald
  • 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）
  • Year and Date: 2016-03-23
• [Presentation] HlyFによるOuter membrane vesicles（外膜小胞）産生誘導機構の解明 (2015)
  • Author(s): 村瀬 一典, Patricia Martin, Jean-Philippe Nougayrede, 林 哲也, Eric Oswald
  • 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] Role for hemolysin F in the virulence of Escherichia coli (2014)
  • Author(s): Martin P, Murase K, Houle S, Helloin E, Nougayrede JP, Dozois C, Hayashi T, Oswald E
  • Organizer: ASM 2014 (114th General Meeting, American Society for Microbiology)
  • Place of Presentation: Boston Convention & Exhibition Center (Boston, Massachusetts)
  • Year and Date: 2014-05-17 – 2014-05-20