限定的発現遺伝子群espが担う新たな細菌特性の解明

• Project/Area Number: 18H02652
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 49050:Bacteriology-related
• Research Institution: University of Tsukuba
• Principal Investigator: 森川 一也 筑波大学, 医学医療系, 教授 (90361328)
• Co-Investigator(Kenkyū-buntansha): 尾花 望 筑波大学, 医学医療系, 助教 (00722688)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000); Fiscal Year 2021: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2020: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2019: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2018: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
• Keywords: 黄色ブドウ球菌 / esp / 不均一性 / 限定的発現 / 限定的発発現 / 限定的発言

Outline of Annual Research Achievements

本研究対象である黄色ブドウ球菌の限定的発現遺伝子esp (expression in minor subpopulation)は数パーセント以下の細胞で限定的に発現するものである。本研究は、esp遺伝子群が生む細胞集団の多様性に基づいた新たな特性を明らかにすることを目標とした。esp発現細胞での遺伝子発現プロフィールを明らかにするために、これまでにesp発現細胞をセルソーターにより回収してRNAseqまで行う実験系を構築してきた。その中でセルソータ実験におけるレポータの蛍光強度や発現頻度のクリアすべき下限が明らかとなり、レポータ強度の問題を解決するための各種レポータを準備した。発現頻度条件をクリアできる培養条件の検討は今後も継続してすすめ、残された機能未知のesp遺伝子の役割を解明する予定である。esp16(com遺伝子)の機能、すなわち細胞外DNAの取り込み、についてさらに研究をすすめ、これがブドウ球菌カセット染色体の取り込みによるMRSA出現のメカニズムであること、またバイオフィルム中で複雑なシグナル伝達系に制御されつつ機能することなどを見出し、多くの追加実験を行って論文として発表する準備を完了した（Nature communications 受理）。また、esp16と同時に発現する遺伝子群を上記の方法で明らかにすることができた。多くのDNA取り込み関連遺伝子群の他、機能未知遺伝子も含まれていた（投稿準備中）。

Research Progress Status

令和3年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和3年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] Biotechnology Center of Ho Chi Minh City(ベトナム)
• [Int'l Joint Research] Institut Pasteur(フランス)
• [Int'l Joint Research] Institut Pasteur(フランス)
• [Int'l Joint Research] Biotechnology Center of Ho Chi Minh City(ベトナム)
• [Int'l Joint Research] Biotechnology Center of Ho Chi Minh City(ベトナム)
• [Int'l Joint Research] Biotechnology Center Ho Chi Minh City(ベトナム)
• [Journal Article] Natural transformation allows transfer of SCCmec-mediated methicillin resistance in Staphylococcus aureus biofilm. (2022)
  • Author(s): Mais Maree, Le Thuy Thi Nguyen, Ryosuke L. Ohniwa, Masato Higashide, Tarek Msadek, Kazuya Morikawa
  • Journal Title: Nature communications Volume: 未定
  • DOI: 10.1038/s41467-022-29877-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] No Change, No Life? What We Know about Phase Variation in Staphylococcus aureus (2021)
  • Author(s): Gor Vishal、Ohniwa Ryosuke L.、Morikawa Kazuya
  • Journal Title: Microorganisms Volume: 9 Pages: 244-244
  • DOI: 10.3390/microorganisms9020244
  • Peer Reviewed / Open Access
• [Journal Article] Finding of Agr phase variants in Staphylococcus aureus. (2019)
  • Author(s): Vishal Gor, Aya J. Takemura, Masami Nishitani, Masato Higashide, Veronica Medrano Romero, Ryosuke L. Ohniwa, and Kazuya Morikawa
  • Journal Title: mBio Volume: 10
  • DOI: 10.1128/mbio.00796-19
  • NAID: 120007132913
  • Peer Reviewed / Open Access
• [Journal Article] Fitness of Spontaneous Rifampicin-resistant Staphylococcus aureus isolates in a biofilm environment. (2019)
  • Author(s): Lisa Maudsdotter, Yuri Ushijima, Kazuya Morikawa
  • Journal Title: Front Microbiol Volume: 10
  • DOI: 10.3389/fmicb.2019.00988
  • NAID: 120007133364
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] What Happens in the Staphylococcal Nucleoid Under Oxidative Stress? (2019)
  • Author(s): Kazuya Morikawa, Yuri Ushijima , Ryosuke L. Ohniwa, Masatoshi Miyakoshi and Kunio Takeyasu
  • Journal Title: Microorganisms Volume: 7 Pages: 631-631
  • DOI: 10.3390/microorganisms7120631
  • NAID: 120007132875
  • Peer Reviewed / Open Access
• [Presentation] 黄色ブドウ球菌Agr系の相変異 (2022)
  • Author(s): 森川一也、Vishal Gor, 東出正人、Veronica Medrano Romero、大庭良介
  • Organizer: 第95回日本細菌学会総会、シンポジウム細菌のリバーシブル変異と適応戦略
• [Presentation] MRSA emergence is mediated by natural transformation. (2021)
  • Author(s): Mais Maree, Le Thuy Thi Nguyen, Ryosuke L. Ohniwa Shenghe Huang, Masato Higashide, Tarek Msadek, Kazuya Morikawa
  • Organizer: World Microbe Forum 2021
  • Int'l Joint Research
• [Presentation] Virulence Reversion in Staphylococcus aureus (2020)
  • Author(s): Vishal Gor, Mitsuaki Hoshi, Aya J Takemura, Masato Higashide, Veronica Medrano Romero, Ryosuke L Ohniwa, Kazuya Morikawa
  • Organizer: 1st International Electronic Conference on Microbiology
  • Int'l Joint Research
• [Presentation] Prevalence of Agr Phase Variants in Staphylococcus aureus (2019)
  • Author(s): Vishal Gor, Mitsuaki Hoshi, Aya J. Takemura, Masato Higashide, Veronica Medrano Romero, Ryosuke L. Ohniwa, and Kazuya Morikawa
  • Organizer: Tsukuba Global Science Week
  • Int'l Joint Research
• [Presentation] Involvement of two-component systems in development in Staphylococcus aureus (2019)
  • Author(s): Mais Maree, Le Thuy Nguyen Thi, Ryosuke L. Ohniwa, and Kazuya Morikawa
  • Organizer: Tsukuba Global Science Week
  • Int'l Joint Research
• [Presentation] Functional analysis of nucleoid dynamics in Staphylococcus aureus. (2019)
  • Author(s): Eunsam Lee, Mais Maree, and Kazuya Morikawa
  • Organizer: The 1st University of Tsukuba- University of Science Joint Young Researcher Meeting in Biomdeical Science
  • Int'l Joint Research
• [Presentation] Report of Two Linezolid-Resistant Staphylococcal Isolates from University Medical Center, Ho Chi Minh City. (2019)
  • Author(s): Nguyen Thi Le Thuy, Nguyen Thi Khanh Ngau, Le Thi Anh Phuc Nhi, Fabio Cafini, Morikawa Kazuya
  • Organizer: The 1st University of Tsukuba- University of Science Joint Young Researcher Meeting in Biomdeical Science
  • Int'l Joint Research
• [Presentation] Can “natural transformation” transmit SCCmec among staphylococci? (2019)
  • Author(s): Mais Maree Nguyen Thi Le Thuy, and Kazuya Morikawa.
  • Organizer: The 1st University of Tsukuba- University of Science Joint Young Researcher Meeting in Biomdeical Science
  • Int'l Joint Research
• [Presentation] 黄色ブドウ球菌における自然形質転換の制御 (2018)
  • Author(s): 宮井拓哉、Nguyen Thi Le Thuy、森川一也
  • Organizer: 第41回 日本分子生物学会年会
• [Remarks] Laboratory of Microbiology
  • URL: https://www.md.tsukuba.ac.jp/basic-med/infectionbiology/microbiology/index-e.html
• [Remarks] 微生物学研究室
  • URL: https://www.md.tsukuba.ac.jp/basic-med/infectionbiology/microbiology/index.html
• [Remarks] 研究室HP研究内容
  • URL: http://www.md.tsukuba.ac.jp/basic-med/infectionbiology/microbiology/research.html
• [Remarks] 筑波大学 感染生物学（微生物学）
  • URL: http://www.md.tsukuba.ac.jp/basic-med/infectionbiology/microbiology/
• [Remarks] 感染生物学微生物学研究室
  • URL: http://www.md.tsukuba.ac.jp/basic-med/infectionbiology/microbiology/