Mechanism of phenotype diversification by bacterial methylome

• Project/Area Number: 18K14672
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 43050:Genome biology-related
• Research Institution: Hokkaido University
• Principal Investigator: Furuta Yoshikazu 北海道大学, 人獣共通感染症国際共同研究所, 講師 (40613667)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2020: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
• Keywords: DNA修飾 / エピゲノム / メチル化酵素 / DNAメチル化酵素 / エピゲノム解析 / DNAメチル化 / メチローム / 制限修飾系 / 遺伝子発現制御 / トランスクリプトーム

Outline of Final Research Achievements

To analyze how DNA methylation affect phenotype of bacterial cells, we aimed to develop an E. coli library with diversity in DNA methylation status by developing DNA methyltransferase enzyme with various DNA methylation target sequence utilizing one-hybrid method and also by searching a DNA methyltransferase with unknown target sequence from databases.
The former didn't work well as it was difficult to obtain an enzyme with ideal activity with enough specificity and biochemical activity. For the latter, we discovered a novel DNA methyltransferase with complex target sequence.

Academic Significance and Societal Importance of the Research Achievements

データベース中のDNAメチル化酵素の活性を探索することで、今回のような複雑なメチル化ターゲット配列を示すものを得られたことによって、バクテリアのメチル化パターンの多様性が示された。今後はこうした複雑なメチル化を施すDNAメチル化酵素の役割や、環境中のバクテリア中のメチル化パターンの多様性について、解析を行っていく。

Research Products

• [Journal Article] A GCDGC-specific DNA (cytosine-5) methyltransferase that methylates the GCWGC sequence on both strands and the GCSGC sequence on one strand (2022)
  • Author(s): Furuta Yoshikazu、Miura Fumihito、Ichise Takahiro、Nakayama Shouta M. M.、Ikenaka Yoshinori、Zorigt Tuvshinzaya、Tsujinouchi Mai、Ishizuka Mayumi、Ito Takashi、Higashi Hideaki
  • Journal Title: PLOS ONE Volume: 17 Issue: 3 Pages: e0265225-e0265225
  • DOI: 10.1371/journal.pone.0265225
  • Peer Reviewed / Open Access
• [Journal Article] Networking and Specificity-Changing DNA Methyltransferases in Helicobacter pylori (2020)
  • Author(s): Yano Hirokazu、Alam Md. Zobaidul、Rimbara Emiko、Shibata Tomoko F.、Fukuyo Masaki、Furuta Yoshikazu、Nishiyama Tomoaki、Shigenobu Shuji、Hasebe Mitsuyasu、Toyoda Atsushi、Suzuki Yutaka、Sugano Sumio、Shibayama Keigo、Kobayashi Ichizo
  • Journal Title: Frontiers in Microbiology Volume: 11 Pages: 1628-1628
  • DOI: 10.3389/fmicb.2020.01628
  • Peer Reviewed / Open Access
• [Presentation] 長鎖型シークエンサーを用いた炭疽菌のゲノム欠失およびDNAメチル化の解析 (2019)
  • Author(s): 古田芳一、播磨勇人、伊藤絵美子、丸山史人、大西なおみ、大崎研、小川寛人、David Squarre、Bernard M. Hang'ombe、東秀明
  • Organizer: 第92回日本細菌学会総会
  • Invited