環境保全微生物に生じたDNAダメージと修復のアダクトミクスに関する研究

2015 年度 実施状況報告書

• 研究課題/領域番号: 26505010
• 研究機関: 横浜市立大学
• 研究代表者: Robert Kanaly 横浜市立大学, 生命ナノシステム科学研究科, 教授 (00448656)
• 研究期間 (年度): 2014-04-01 – 2017-03-31
• キーワード: DNA adductomics

研究実績の概要

Exposure assays with hydrogen peroxide were conducted using the model organism. DNA adducts were identified in bacterial DNA. Relative levels of some adducts were measured and differences were observed when comparing exposed and unexposed organisms. Following, exposure assays with model organism and PAH quinone, 1,2-naphthoquinone were conducted. DNA adducts were identified. Possible exposure-specific adducts of guanine were detected. Considering these results, in vitro reactions of 1,2-naphthoquinone and 2’-deoxyguanosine dG were conducted. Products were analyzed by LC-MS/MS to aid in identification of adducts in bacterial DNA. Research is ongoing.

現在までの達成度 (区分)

3: やや遅れている

理由

Model organism was decided and production of PAH-type catechols during biotransformation of PAHs was confirmed by LC/ESI-MS/MS. Genes involved in catabolism of PAHs by this organism were determined and pathways for the biotransformation of model PAHs were mapped. RT-PCR analyses were conducted as part of methods development. Assessment of DNA damage by LC/ESI-MS/MS DNA adductomics was conducted using the model organism and model toxicant hydrogen peroxide. Afterwards, DNA damage assessment was conducted by using 1,2-naphthoquinone. DNA adducts were identified in both conditions. Exposure specific adduct(s) of guanine appeared to have occurred after exposure to 1,2-naphthoquinone. In vitro reactions of 1,2-naphthoquinone and 2’-deoxyguanosine dG were conducted with the aim to identify DNA adducts in bacterial DNA.

今後の研究の推進方策

Continue DNA damage analyses using 1,2-naphthoquinone. Develop methods for conducting transcriptomics on the model organism. Conduct transcriptome analyses. Develop methods for detecting ROS by fluorescence in the model organism.

次年度使用額が生じた理由

Budget from 2015-16 was carried over to 2016-17 because it was sufficient for the experiments conducted in 2015-16.

次年度使用額の使用計画

Continuation of exposure assays by using LC/ESI-MS/MS. Transcriptomics analyses. Presentation of results at International peer review conference.

研究成果

• [雑誌論文] Analysis of the Fine-Tuning of Cyanobacterial Circadian Phase by Monochromatic (2016)
  • 著者名/発表者名: Kobayashi T, Obana Y, Kuboi N, Kitayama Y, Hayashi S, Oka M, Wada N, Arita K, Shimizu T, Sato M, Kanaly RA, Kutsuna S
  • 雑誌名: Plant Cell Physiol 巻: 57 ページ: 105-114
  • DOI: 10.1093/pcp/pcv177
  • 査読あり / オープンアクセス / 国際共著 / 謝辞記載あり
• [雑誌論文] Application of DNA adductomics to soil bacterium Sphingobium sp. strain KK22 (2015)
  • 著者名/発表者名: Kanaly, R. A., R. Micheletto, T. Matsuda, Y. Utsuno, Y. Ozeki and N. Hamamura
  • 雑誌名: MicrobiologyOpen 巻: 4 ページ: 841-856
  • DOI: 10.1002/mbo3.283
  • 査読あり / オープンアクセス / 国際共著 / 謝辞記載あり
• [雑誌論文] Sphingobium barthaii sp. nov., a high molecular weight polycyclic aromatic hydrocarbon-degrading bacterium isolated from cattle pasture soil (2015)
  • 著者名/発表者名: Maeda, A.H., M. Kunihiro, Y. Ozeki, Y. Nogi and R.A. Kanaly
  • 雑誌名: International Journal of Systematic and Evolutionary Microbiology 巻: 65 ページ: 2919-2924
  • DOI: 10.1099/ijs.0.000356
  • 査読あり / 国際共著 / 謝辞記載あり
• [雑誌論文] Aerobic biotransformation of 3-methylindole to ring cleavage products by (2015)
  • 著者名/発表者名: Fukuoka K, Ozeki Y, Kanaly RA
  • 雑誌名: Biodegradation 巻: 26 ページ: 359-373
  • DOI: 10.1007/s10532-015-9739-0
  • 査読あり / 国際共著 / 謝辞記載あり
• [雑誌論文] MytiLec, a mussel R-type lectin, interacts with surface glycan Gb3 on Burkitt’s lymphoma cells to trigger apoptosis through multiple pathways (2015)
  • 著者名/発表者名: Imtiaj Hasan, Shigeki Sugawara, Yuki Fujii, Yasuhiro Koide, Daiki Terada, Naoya Iimura, Toshiyuki Fujiwara, Keisuke G. Takahashi, Nobuhiko Kojima, Sultana Rajia, Sarkar M. A. Kawsar, Robert A. Kanaly, Hideho Uchiyama, Masahiro Hosono, Yukiko Ogawa, Hideaki Fujita, Jiharu Hamako, Taei Matsui, Yasuhiro Ozeki
  • 雑誌名: Marine Drugs 巻: 13 ページ: 7377-7389
  • DOI: 10.3390/md13127071
  • 査読あり / オープンアクセス / 国際共著 / 謝辞記載あり
• [雑誌論文] Expression of various types of glycans in the lugworm (Perinereis nuntia var. vallata) (2015)
  • 著者名/発表者名: Koide Y, Hasan I, Asanuma A, Fujii Y, Ogawa Y, Kobayashi H, Rajia S, Kawsar SMA, Kanaly RA, Ozeki Y.
  • 雑誌名: Annals of Marine Biology and Research 巻: 2 ページ: 1005
  • 査読あり / オープンアクセス / 国際共著 / 謝辞記載あり
• [学会発表] Investigation of DNA adducts in PAH-degrading bacterial cells after exposure to hydrogen peroxide by DNA adductomics (2015)
  • 著者名/発表者名: Fukuoka, K., and R.A. Kanaly
  • 学会等名: The 44th Annual Meeting of the Japanese Environmental Mutagen Society
  • 発表場所: Fukuoka, Japan
  • 年月日: 2015-11-27 – 2015-11-28
• [学会発表] MytiLec, an R-type lectin from Mediterranean mussel regulates cancer cell growth through glycan-dependent signaling. (2015)
  • 著者名/発表者名: Hasan I, Fujii Y, Ogawa Y, Sugawara S, Hosono M, Koide Y, Rajia S, Kawsar S M A, Kanaly R A, Fujiwara T, Fujita H, Ozeki Y
  • 学会等名: 23rd International Glycoconjugates Symposium (GLYCO23)
  • 発表場所: Split, Croatia
  • 年月日: 2015-09-15 – 2015-09-20
  • 国際学会
• [学会発表] Analysis of DNA damage in bacterial cells by DNA adductomics (2015)
  • 著者名/発表者名: Kanaly, R A., K. Fukuoka, and R. Micheletto
  • 学会等名: International Congress of the Asian Society of Toxicology (ASIATOX 2015)
  • 発表場所: Jeju, South Korea
  • 年月日: 2015-06-23 – 2015-06-26
  • 国際学会
• [図書] Chapter 33, "Structural glycobiology for lectin to promote advanced biomedical research" Marine Glycobiology, Principles and Applications. (Edited by Se-Kwon Kim, ISBN: 9781498709613) (2016)
  • 著者名/発表者名: 1. Hasan I, Fujii Y, Rajia S, Kawsar S M A, Sugawara S, Ogawa Y, Hosono M, Koide Y, Kanaly R A, Ozeki Y.
  • 総ページ数: 443-458
  • 出版者: Taylor & Francis Group, CRC press
• [図書] Chapter 21: SUEL/RBL-Lectins and their biomedical applications" Marine Omics, Principles and Applications (Se-Kwon Kim ed, ISBN 9781482258202) (2016)
  • 著者名/発表者名: 5. Koide Y, Hasan I, Fujii Y, Kawsar SMA, Rajia S, Sugawara S, Ogawa Y, Hosono M, Kanaly RA, Ozeki Y.
  • 総ページ数: 407-418
  • 出版者: Taylor & Francis Group, CRC press