九州大学 研究者情報
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基本情報 研究活動 教育活動
石野 園子(いしの そのこ) データ更新日:2019.06.24

准教授 /  農学研究院 生命機能科学部門 生物機能分子化学講座


主な研究テーマ
DNA複製、組換え、修復の分子機構解析
キーワード:DNA複製  DNA修復  DNA組換え 
2008.04~2019.03.
研究業績
主要原著論文
1. Ying Liu, David Brandt, Sonoko Ishino, Yoshizumi Ishino, Eugene V. Koonin, Jörn Kalinowski, Mart Krupovic, David Prangishvili, New archaeal viruses discovered by metagenomic analysis of viral communities in enrichment cultures, Environmental Microbiology, 10.1111/1462-2920.14479, 2019.01, [URL], Viruses infecting hyperthermophilic archaea of the phylum Crenarchaeota display enormous morphological and genetic diversity, and are classified into 12 families. Eight of these families include only one or two species, indicating sparse sampling of the crenarchaeal virus diversity. In an attempt to expand the crenarchaeal virome, we explored virus diversity in the acidic, hot spring Umi Jigoku in Beppu, Japan. Environmental samples were used to establish enrichment cultures under conditions favouring virus replication. The host diversity in the enrichment cultures was restricted to members of the order Sulfolobales. Metagenomic sequencing of the viral communities yielded seven complete or near-complete double-stranded DNA virus genomes. Six of these genomes could be attributed to polyhedral and filamentous viruses that were observed by electron microscopy in the enrichment cultures. Two icosahedral viruses represented species in the family Portogloboviridae. Among the filamentous viruses, two were identified as new species in the families Rudiviridae and Lipothrixviridae, whereas two other formed a group seemingly distinct from the known virus genera. No particle morphotype could be unequivocally assigned to the seventh viral genome, which apparently represents a new virus type. Our results suggest that filamentous viruses are globally distributed and are prevalent virus types in extreme geothermal environments..
2. Sonoko Ishino, Stéphane Skouloubris, Hanae Kudo, Caroline L'Hermitte-Stead, Asmae Es-Sadik, Jean Christophe Lambry, Yoshizumi Ishino, Hannu Myllykallio, Activation of the mismatch-specific endonuclease EndoMS/NucS by the replication clamp is required for high fidelity DNA replication, Nucleic acids research, 10.1093/nar/gky460, 46, 12, 6206-6217, 2018.07, [URL], The mismatch repair (MMR) system, exemplified by the MutS/MutL proteins, is widespread in Bacteria and Eukarya. However, molecular mechanisms how numerous archaea and bacteria lacking the mutS/mutL genes maintain high replication fidelity and genome stability have remained elusive. EndoMS is a recently discovered hyperthermophilic mismatch-specific endonuclease encoded by nucS in Thermococcales. We deleted the nucS from the actinobacterium Corynebacterium glutamicum and demonstrated a drastic increase of spontaneous transition mutations in the nucS deletion strain. The observed spectra of these mutations were consistent with the enzymatic properties of EndoMS in vitro. The robust mismatch-specific endonuclease activity was detected with the purified C. glutamicum EndoMS protein but only in the presence of the β-clamp (DnaN). Our biochemical and genetic data suggest that the frequently occurring G/T mismatch is efficiently repaired by the bacterial EndoMS-β'clamp complex formed via a carboxy-terminal sequence motif of EndoMS proteins. Our study thus has great implications for understanding how the activity of the novel MMR system is coordinated with the replisome and provides new mechanistic insight into genetic diversity and mutational patterns in industrially and clinically (e.g. Mycobacteria) important archaeal and bacterial phyla previously thought to be devoid of the MMR system..
3. Mariko Nagata, Sonoko Ishino, Takeshi Yamagami, Hiromi Ogino, Jan Robert Simons, Tamotsu Kanai, Haruyuki Atomi, Yoshizumi Ishino, The Cdc45/RecJ-like protein forms a complex with GINS and MCM, and is important for DNA replication in Thermococcus kodakarensis, Nucleic acids research, 10.1093/nar/gkx740, 45, 18, 10693-10705, 2017.10, [URL], The archaeal minichromosome maintenance (MCM) has DNA helicase activity, which is stimulated by GINS in several archaea. In the eukaryotic replicative helicase complex, Cdc45 forms a complex with MCM and GINS, named as CMG (Cdc45-MCM-GINS). Cdc45 shares sequence similarity with bacterial RecJ. A Cdc45/RecJ-like protein from Thermococcus kodakarensis shows a bacterial RecJ-like exonuclease activity, which is stimulated by GINS in vitro. Therefore, this archaeal Cdc45/RecJ is designated as GAN, from GINS-associated nuclease. In this study, we identified the CMG-like complex in T. kodakarensis cells. The GANGINS complex stimulated the MCM helicase, but MCM did not affect the nuclease activity of GAN in vitro. The gene disruption analysis showed that GAN was non-essential for its viability but the Agan mutant did not grow at 93 C. Furthermore, the Agan mutant showed a clear retardation in growth as compared with the parent cells under optimal conditions at 85 C. These deficiencies were recovered by introducing the gan gene encoding the nuclease deficient GAN protein back to the genome. These results suggest that the replicative helicase complex without GAN may become unstable and ineffective in replication fork progression. The nuclease activity of GAN is not related to the growth defects of the δgan mutant cells..
4. Ying Liu, Sonoko Ishino, Yoshizumi Ishino, Gérard Pehau-Arnaudet, Mart Krupovic, David Prangishvili, A novel type of polyhedral viruses infecting hyperthermophilic archaea, Journal of Virology, 10.1128/JVI.00589-17, 91, 13, 2017.07, [URL], Encapsidation of genetic material into polyhedral particles is one of the most common structural solutions employed by viruses infecting hosts in all three domains of life. Here, we describe a new virus of hyperthermophilic archaea, Sulfolobus polyhedral virus 1 (SPV1), which condenses its circular double-stranded DNA genome in a manner not previously observed for other known viruses. The genome complexed with virion proteins is wound up sinusoidally into a spherical coil which is surrounded by an envelope and further encased by an outer polyhedral capsid apparently composed of the 20-kDa virion protein. Lipids selectively acquired from the pool of host lipids are integral constituents of the virion. None of the major virion proteins of SPV1 show similarity to structural proteins of known viruses. However, minor structural proteins, which are predicted to mediate host recognition, are shared with other hyperthermophilic archaeal viruses infecting members of the order Sulfolobales. The SPV1 genome consists of 20,222 bp and contains 45 open reading frames, only one-fifth of which could be functionally annotated..
主要学会発表等
1. 石野園子、工藤華枝、Stephane Skouloubris、Caroline l’Hermitte-Stead、Asmae Essadik、Jean Christophe Lambry、Hannu Myllykallio、石野良純, ミスマッチ特異的エンドヌクレアーゼEndoMS/NucSのDNA修復における役割, 第41回日本分子生物学会年会, 2018.11.
2. Sonoko Ishino, Ken-ichi Miyazono, Naruto Makita, Tomoko Ito, Masaru Tanokura and Yoshizumi Ishino., Structure-based mutational analysis of the lesion-specific endonuclease., Molecular Biology of Archaea 6 / EMBO Workshop, 2018.09.
3. Sonoko Ishino, Kouta Mayanagi, Tsuyoshi Shirai, Shinichi Kiyonari, Namiko Imai, Takuji Oyama, and Yoshizumi Ishino, Biochemical and Structural analyses of replication proteins bound to PCNA., 12th International Congress on Extremophiles, 2018.09.
学会活動
学会大会・会議・シンポジウム等における役割
2018.12.08~2018.12.09, 極限環境生物学会, 評議委員.
その他の研究活動
海外渡航状況, 海外での教育研究歴
Shandong Univ., China, 2018.12~2018.12.
ウィーン大学, パスツール研究所, ナポリ大学, Austria, France, Italy, 2018.09~2018.09.
研究資金
科学研究費補助金の採択状況(文部科学省、日本学術振興会)
2018年度~2020年度, 基盤研究(C), 代表, 超好熱性アーキアから発見された新規エンドヌクレアーゼによるDNA修復経路の解明.

九大関連コンテンツ

pure2017年10月2日から、「九州大学研究者情報」を補完するデータベースとして、Elsevier社の「Pure」による研究業績の公開を開始しました。
 
 
九州大学知的財産本部「九州大学Seeds集」