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DNA Stable-Isotope Probing DNA-SIP
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静止段階の細菌におけるグローバルDNA圧縮は転写に影響を与えない
Richard Janissen1, Mathia M A Arens1, Natalia N Vtyurina1
1Department of Bionanoscience, Kavli Institute of Nanoscience, Delft, South-Holland 2629HZ, the Netherlands.
Cell
|July 31, 2018
まとめ
飢えた細胞からのDNA結合タンパク質 (Dps) は,大腸菌のDNAを凝縮し,それを保護します. DpsはDNA結合タンパク質を排除しながら,RNAポリメラーゼが動的に遺伝子にアクセスすることを可能にします.
科学分野:
- 微生物学
- 分子生物学
- 生物化学
背景:
- 飢えた細胞からのDNA結合タンパク質 (Dps) は静止段階のEscherichia coliに豊富に含まれています.
- DpsはDNAを圧縮し,損傷から保護し,グローバルな転写調節剤として提案されています.
- RNAポリメラーゼ (RNAP) 活動に対するDps誘発のDNA圧縮の効果は,直接的な検査を必要とする.
研究 の 目的:
- E. coli の RNAP 活性に対する Dps 媒介の DNA 圧縮の影響を調査する.
- DNAへの RNAP アクセスを変えて Dps が転写を調節するかどうかを判断する.
- Dps,DNA,RNAPのダイナミックな相互作用を特徴づけること.
主な方法:
- ニュクレオイド構造と全体的な遺伝子発現を評価するための遺伝子削除研究.
- DNAとタンパク質との相互作用をテストするインビトロ結合測定法
- 単一分子アッセイは,RNAPの延長に関するDpsダイナミクスを観察する.
主要な成果:
- dps遺伝子を削除すると,核細胞が分解されるが,トランスクリプトームとプロテオームに最小の影響を及ぼした.
- In vitroでは,Dpsは制限酵素を阻害したが,RNAPはDNA結合を阻害しなかった.
- 単一分子測定では,DpsがRNAPの進行を阻害することなくDNAを動的に凝縮することを示した.
結論:
- Dpsは,特定のDNA結合タンパク質を除外するダイナミックな構造を形成しますが,RNAPが遺伝子へのアクセスを許可します.
- この振る舞いは相分離オーガネルのものと類似している.
- DpsのDNA圧縮における役割は,静止期E. coliにおける転写を全体的に制御しない.
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